medical research with mice

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Published: 05 December 2002

Mice make medical history

Tom Clarke

Nature ( 2002 ) Cite this article

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The scientific credentials of the lab mouse are impressive.

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Clarke, T. Mice make medical history. Nature (2002). https://doi.org/10.1038/news021202-10

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Published : 05 December 2002

DOI : https://doi.org/10.1038/news021202-10

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NIH to make a mightier mouse resource for understanding disease

Over the next five years, National Institutes of Health (NIH)-funded researchers will extensively test and generate data about mice with disrupted genes to gain clues about human diseases. NIH today awarded a set of cooperative agreements totaling more than $110 million to begin the second phase of the Knockout Mouse Project (KOMP).

The results of the next stage, called the Knockout Mouse Phenotyping Project, or KOMP2, will be placed in a public database. Researchers make knockout mice by disrupting the function of individual genes across the animal’s genome.

KOMP2 is a trans-NIH and NIH Common Fund project that will work with other members of the International Knockout Mouse Phenotyping Consortium (IMPC) to generate about 5,000 strains of knockout mice that will undergo a large battery of clinical phenotype tests. A phenotype includes biological information about appearance, behavior and other measurable physical and biochemical characteristics. Such information will help reveal how all traits are affected by deleting a given gene in an individual mouse.

In the long term, the project aims to enable the research community to establish the traits associated with the function of every protein-coding gene in the mammalian genome. Such information will be valuable for the discovery of the genetic causes of human diseases and will aid efforts to identify new drug targets.

"The generation of detailed phenotypic information for each knockout mouse strain will be a boon to disease researchers who want to determine the function of genes and to improve mouse models of human disease," said NIH Director Francis S. Collins, M.D., Ph.D. "I am grateful to all of the people and programs across NIH who are supporting this effort and to our international partners who have joined us in this scientific endeavor."

In partnership with several international programs, the initial five-year phase of KOMP will reach its goal of creating knockout mouse embryonic stem cell lines for each of the approximately 21,000 protein-coding genes in the mouse genome this year. The International Knockout Mouse Consortium (IKMC) includes the Knockout Mouse Project (KOMP), U.S.A.; the European Conditional Mouse Mutagenesis Program (EUCOMM) funded by the European Commission: the Texas A&M Institute for Genomic Medicine (TIGM); and the North American Conditional Mouse Mutagenesis Project (NorCOMM) funded by Genome Canada.

"NIH is committed to making knockout mouse models more widely accessible to the biomedical research community," said James Battey, M.D., Ph.D., director of the National Institute on Deafness and Other Communication Disorders (NIDCD), who is also a co-chairman of the Trans-NIH Mouse Initiative. "Getting these valuable models into the hands of a wide range of researchers will serve to accelerate our efforts to develop new strategies for understanding and treating human disease."

During the next five years, KOMP2 will transform the knockout mouse embryonic stem (ES) cells into adult mice for 2,500 lines of well-characterized knockout mice strains, and IMPC will create about 2,500 additional knockout mouse strains. Each mouse will undergo the same standard analysis so that the results can be compared for all of the mice tested. NIH has awarded six cooperative agreements to three groups to establish production and phenotype centers for the project.

"It is going to take a great deal of scientific teamwork to assimilate phenotypic information about this knockout mouse resource, but we are confident in the team that has been assembled to accomplish the task," said National Human Genome Research Institute (NHGRI) Director Eric D. Green, M.D., Ph.D. NHGRI is involved in the planning and administration of KOMP2.

The National Center for Research Resources (NCRR) will administer the awards for the production centers, and NHGRI will administer the awards for the phenotyping centers. NCRR and NHGRI are components of the NIH.

The funded groups will all receive a total of approximately $34 million and are expected to produce and phenotype 833 strains of knockout mice each for a total of about 2,500 knockout mouse lines. Recipients of the awards are:

Baylor College of Medicine, Houston. This center will collaborate with the Wellcome Trust Sanger Institute in Hinxton, England and the Medical Research Council (MRC) Harwell in Oxfordshire, England.
University of California, Davis. This center will collaborate with the Toronto Center for Phenogenomics in Canada, Children’s Hospital Oakland Research Institute in California, and Charles River Laboratories in Wilmington, Mass.
The Jackson Laboratory in Bar Harbor, Maine.

"This resource will enable many more researchers to tap into the power of knockout mice for exploring gene function, which in turn will speed our efforts to improve human health," said Louise E. Ramm, Ph.D., acting director, National Center for Research Resources.

In addition to the production and phenotype centers, NIH awarded a five-year, cooperative agreement totaling $10 million to the European Bioinformatics Institute in Hinxton, England, which will collaborate with MRC Harwell and Wellcome Trust Sanger Institute to set up a data coordination center and database to track progress of the project and to coordinate efforts between KOMP2 and IMPC researchers. In addition, this center will build an integrated Web portal that will provide researchers access to the phenotype data.

The mouse is a key mammalian system in which to produce a genomics resource because of the long history and depth of understanding of mouse genetics and the availability of the mouse genome sequence. What's more, researchers have made advances over the last several years in improving the efficiency and decreasing the cost of generating knockout mice.

Historically, researchers have generated their own lines of knockout mice to serve as models for human disease, such as heart disease or cancer. However, rather than generating a detailed and comprehensive phenotype of the mouse, they often are only interested in a handful of phenotypes. For example, a researcher interested in cardiovascular disease may only want to examine the effect of a disrupted gene on blood pressure.

This single-lab approach can be expensive and inefficient. A researcher with access to a low-cost knockout mouse that has been extensively phenotyped can focus his or her time and research budget on more in-depth research questions rather than spending it on producing a knockout mouse about which the researcher has limited information.

KOMP2 and IMPC researchers will begin by creating lines of knockout mice from embryonic stem cells produced by KOMP. The 5,000 genes that will be knocked out will be selected from nominations already submitted by the research community. Many of the selected genes will be used to study disease processes and underlying mechanisms. Others will be selected based on the genetic variations associated with the human diseases that have been uncovered by genome-wide association studies.

Statistically, about 25 percent of the mouse pups will inherit both copies of the knocked out gene, while their littermates will have only one copy and be heterozygous, or normal. The knockout mice and the healthy littermates will both undergo a battery of more than 400 phenotype measurements at multiple times during their lives. Tests will include X-ray imaging, magnetic resonance imaging (MRI), blood exams, balance tests, and urine and fecal analysis, to name a few. Both the knockout and normal phenotype data will be made available through the KOMP2 data coordination center so that researchers who acquire and study the knockout mice can compare various phenotypes.

"We want to characterize each line of mice broadly with no assumptions about what the gene is or is not doing," said IMPC Executive Director Mark Moore, Ph.D. "If you think of the function of a gene as a needle in a haystack, we’re removing the haystack so you can see what the needle does."

At the end of the initial five years of the effort, the NIH and IMPC will evaluate the usefulness of the resource to the research community. If the evaluation is a positive one, both efforts may scale up to create and phenotype a total of 12,000 more knockout mice.

Once each knockout mouse is phenotyped, researchers can obtain information on what knockout mouse lines are available and how to order them from the University of California Davis KOMP Repository .

To access the IKMC Web portal, please go to www.knockoutmouse.org .

The 18 NIH institutes, centers and offices contributing to the Knockout Mouse Project are: the NIH Office of Strategic Coordination/Common Fund; NCRR; the National Eye Institute; NHGRI; the National Heart, Lung and Blood Institute; the National Institute on Aging; the National Institute of Alcohol Abuse and Alcoholism; the National Institute of Arthritis and Musculoskeletal and Skin Diseases; the Eunice Kennedy Shriver National Institute of Child Health and Human Development; NIDCD; the National Institute of Dental and Craniofacial Research; the National Institute of Environmental Health Sciences; the National Institute of General Medical Sciences; the National Institute of Mental Health; the National Institute of Neurological Disorders and Stroke; the National Institute of Diabetes and Digestive and Kidney Diseases; the National Cancer Institute; and the Office of AIDS Research.

For more information on the Knockout Mouse Project, go to the NIH Knockout Mouse Project . For a fact sheet describing what knockout mice are, how they are made and what they are used for, go to Knockout Mice . To download a high-resolution photo of knockout mice, go to www.genome.gov/pressDisplay.cfm?photoID=5006 . For more information on the IMPC, go to www.mousephenotype.org/index.html .

The National Center for Research Resources (NCRR), a part of NIH, provides laboratory scientists and clinical researchers with the resources and training they need to understand, detect, treat and prevent a wide range of diseases. NCRR supports all aspects of translational and clinical research, connecting researchers, patients and communities across the nation. For more information, visit www.ncrr.nih.gov .

NHGRI is one of the 27 institutes and centers at NIH. The NHGRI Division of Extramural Research supports grants for research and training and career development at sites nationwide. Additional information about NHGRI can be found at www.genome.gov .

The NIH Common Fund encourages collaboration and supports a series of exceptionally high impact, trans-NIH programs. Programs funded through the Common Fund are managed by the NIH Office of the Director’s Office of Strategic Coordination in partnership with the various NIH Institutes, Centers and Offices. Common Fund programs are designed to pursue major opportunities and gaps in biomedical research that the agency as a whole should address to make the biggest impact possible on the progress of medical research. Additional information about the NIH Common Fund can be found at http://commonfund.nih.gov .

About the National Institutes of Health (NIH): NIH, the nation's medical research agency, includes 27 Institutes and Centers and is a component of the U.S. Department of Health and Human Services. NIH is the primary federal agency conducting and supporting basic, clinical, and translational medical research, and is investigating the causes, treatments, and cures for both common and rare diseases. For more information about NIH and its programs, visit www.nih.gov .

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The Applicability of Mouse Models to the Study of Human Disease

Affiliations.

1 Lung Biology Group, Lund, Sweden.
2 Department of Experimental Medical Science, Lund University, Lund, Sweden.
3 School of Life and Health Sciences, Aston University, Birmingham, UK. [email protected].
PMID: 30788814
PMCID: PMC7121329
DOI: 10.1007/978-1-4939-9086-3_1

The laboratory mouse Mus musculus has long been used as a model organism to test hypotheses and treatments related to understanding the mechanisms of disease in humans; however, for these experiments to be relevant, it is important to know the complex ways in which mice are similar to humans and, crucially, the ways in which they differ. In this chapter, an in-depth analysis of these similarities and differences is provided to allow researchers to use mouse models of human disease and primary cells derived from these animal models under the most appropriate and meaningful conditions.Although there are considerable differences between mice and humans, particularly regarding genetics, physiology, and immunology, a more thorough understanding of these differences and their effects on the function of the whole organism will provide deeper insights into relevant disease mechanisms and potential drug targets for further clinical investigation. Using specific examples of mouse models of human lung disease, i.e., asthma, chronic obstructive pulmonary disease, and pulmonary fibrosis, this chapter explores the most salient features of mouse models of human disease and provides a full assessment of the advantages and limitations of these models, focusing on the relevance of disease induction and their ability to replicate critical features of human disease pathophysiology and response to treatment. The chapter concludes with a discussion on the future of using mice in medical research with regard to ethical and technological considerations.

Keywords: Disease; Ethics; Genetics; Immunology; Model; Mouse; Physiology.

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Mice are the most commonly used animals in medical research. This trend looks likely to continue now that both mouse and human genomes have been mapped ( 80% of human genes are exactly the same as those found in mice, and at least a further 10% are very similar) allowing human genetic disorders and diseases to be studied with greater accuracy.

Often, the only way of determining the function of a human gene is to insert it into, or remove it from, the mouse genome. Many thousands of mouse strains now exist, some frozen as embryos. Eventually, such techniques could lead to new methods of preventing, treating or even curing genetic diseases and other diseases with a genetic component.

Around 87% of all genetically modified animals used in research in the UK are mice [ UK 2020 figures ].

View this post on Instagram A post shared by Understanding Animal Research (@understandinganimalresearch)

Mice ( Mus musculus ) belong to the family rodentia (rodents) and are one of the most common mammals on Earth. They are small animals that grow to the size of 12cm long. Mice are omnivores, their diet consists of a mixture of both plant and animal matter, essentially mice can eat anything they like! Mice have been used in research for more than a century with the first use of mice in genetics dating back to 1902. They are the most commonly used animal in Great Britain.

Why are mice used in research?

Mice have many characteristics that make them ideal laboratory animals. Firstly, some diseases are modelled well in mice as human and mice share some anatomical, physiological, and genetic features. Conveniently, due to the successful sequencing of the mice genome, scientists can produce genetically modified mice and introduce or remove particular genetic features to make the mice better disease models. Mice have a relatively short gestation period and have multiple births, allowing researchers access to a lot of mice in a short amount of time. Their relatively fast ageing process also makes them great models for studying the effects and process of ageing. Finally, b ecause of their size, they are convenient for researchers and animal technicians to house and care for.

What types of research are mice used in?

Mice are versatile, they are used in a range of research from genetics to virology, oncology and many more. Notably, mice and other animals have been very important in the development of Herceptin, a monoclonal antibody used in certain types of breast cancer. Herceptin was the first monoclonal antibody successfully used to treat cancer. The HER2 protein, which makes breast cancer cells grow and duplicate was discovered in rat tumours, however, years later the monoclonal antibodies were used in mice to target the HER2, which successfully reduced tumour growth. The protein was discovered in rat tumours and years later, the antibodies were used to target the HER2 in mice. Herceptin is a humanised mice antibody, being 95% human and 5% mice.

More recently, mice have been instrumental in the search for a coronavirus vaccine. Researcher’s ability to genetically modify mice has been especially useful in breeding mice that are susceptible to the SAR-COV-2 virus. The mice ACE2 receptor is different from the human ACE2 receptor, exempting mice from being infected by the virus. Scientists altered the mice ACE2 receptor to become more human and allow the mice to get the virus and display symptoms of the covid-19 disease. When research began for the vaccine for the virus, mice with humanised ACE2 receptors were the best animal model at researchers’ disposal.

How are the mice looked after?

The use of animals in research is highly reg ulated, an important part of that regulation is ensuring the animals are housed and cared for correctly. Laboratory mice are housed similarly to pet mice, in cages lined with soft, absorbent bedding. In the laboratory the cages are made of see-through plastic so that they can be seen without disturbing them. Mice are given fresh food and water each day and are usually fed a specially constructed diet that meets all their nutritional needs. It is also important that animals have enrichment (things to entertain them), so they will usually have areas where they can hide away and objects to climb and gnaw on.

Because mice are highly social animals, it is very important that they are housed in groups, or at a minimum pairs. There are only a few exceptional circumstances where mice would be kept alone, usually for their own safety.

Find out more about mice in research with our '10 facts' infographic .

See also our pages on GM mice and breeding , mice and stem cell research , our video of mice, and animal research.info on mice and GM mice .

Mice in research: https://www.yourgenome.org/facts/why-use-the- mouse -in-research

Herceptin: https://www.understandinganimalresearch.org.uk/news/herceptin-first-monoclonal-antibody-treatment-for-cancer

Coronavirus and mice: https://www.sciencemag.org/news/2020/04/mice-hamsters-ferrets-monkeys-which-lab-animals-can-help-defeat-new-coronavirus

Mice: https://www.britannica.com/animal/ mouse -rodent/Geographic-distribution-and-habitat

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Why we need female mice in neuroscience research

Catherine Caruso

HMS Communications

Findings reveal that despite hormonal fluctuations, female mice exhibit more stable exploratory behavior than their male peers

Mice have long been a central part of neuroscience research, providing a flexible model that scientists can control and study to learn more about the intricate inner workings of the brain. Historically, researchers have favored male mice over female mice in experiments, in part due to concern that the hormone cycle in females causes behavioral variation that could throw off results.

But new research from Harvard Medical School challenges this notion and suggests that for many experiments, the concern may not be justified.

The study results, published March 7 in Current Biology , reveal that female mice, despite ongoing hormonal fluctuations, exhibit exploratory behavior that is more stable than that of their male peers.

Using a strain of mice commonly studied in lab settings, the researchers analyzed how the animals behaved as they freely explored an open space. They found that the hormone cycle had a negligible effect on behavior and that differences in behavior between individual female mice were much greater. Moreover, differences in behavior were even greater for males than for females, both within and between mice.

The results underscore the importance of incorporating both sexes into mouse studies, the research team said.

“I think this is really powerful evidence that if you’re studying naturalistic, spontaneous exploratory behavior, you should include both sexes in your experiments — and it leads to the argument that in this setting, if you can only pick one sex to work on, you should actually be working on females,” said Sandeep Robert Datta , professor of neurobiology in the Blavatnik Institute at HMS, who co-led the study with Rebecca Shansky of Northeastern University.

From rodents to humans: A history of bias

As neuroscientists strive to better understand the human brain, they routinely turn to the mouse, which Datta considers “the flagship vertebrate model for understanding how the brain works.”

This is because mouse and human brains share a considerable amount of structural organization and genetic information, so scientists can easily manipulate the mouse genome to address specific experimental questions and to build models of human diseases.

“Much of what we understand about the relationship between genes and neural circuits, and between neural activity and behavior, comes from basic research in the mouse, and mouse models are likely going to be really central tools in our fight against a broad array of neurological and psychological diseases,” Datta said.

“I think this is really powerful evidence that if you’re studying naturalistic, spontaneous exploratory behavior, you should include both sexes in your experiments — and it leads to the argument that in this setting, if you can only pick one sex to work on, you should actually be working on females.” Sandeep Robert Datta, professor of neurobiology in the Blavatnik Institute

For more than 50 years, researchers have preferentially used male mice in experiments, and nowhere has this practice been more prominent than in neuroscience. In fact, a 2011 analysis found that there were over five times as many single-sex neuroscience studies of male mice than of female mice. Over time, this practice has resulted in a poorer understanding of the female brain, likely contributing to the misdiagnosis of mental and neurological conditions in women, as well as the development of drugs that have more side effects for women — issues outlined by Shansky in a 2021 perspective in Nature Neuroscience .

The disparity in sex representation common in animal research has also been historically mirrored in research involving human subjects.

“This bias starts in basic science, but the repercussions are rolled into drug development, and lead to bias in drugs being produced, and how drugs are suited for the different sexes,” said lead author Dana Levy , a research fellow in neurobiology at HMS. For example, Levy noted that conditions such as anxiety, depression, and pain are known to manifest differently in female mice and women than in the male mice that are more often used in early-stage drug testing.

To address the problem of sex bias in scientific research, the National Institutes of Health published a policy in 2016 requiring researchers to include male and female subjects and samples in experiments. However, follow-up studies that look across scientific disciplines and examine neuroscience specifically indicate that progress has been slow.

The reasons for such a long-standing bias in neuroscience are complicated, Datta said: “Part of it is just plain old sexism, and part of it is conservatism in the sense that people have studied male mice for so long that they don’t want to make a change.”

Yet perhaps the biggest reason for excluding female mice, Datta said, stems from a widespread assumption that their behavior is broadly affected by cyclic variations in hormones such as estrogen and progesterone — the rodent version of a menstrual cycle, known as the estrous cycle. According to Datta and Levy, estrous status is known to have a strong effect on certain social and sexual behaviors in mice. However, data on the influence of estrous status in other behavioral contexts have been mixed, resulting in what Datta calls “a genuine disagreement in the literature.”

“We wanted to measure how much the estrous cycle seemed to influence basic patterns of exploration,” Datta said. “Our question was whether these ongoing changes in the hormonal state of the mouse affect other neural circuits in a way that’s confusing for researchers.”

“We assumed, like everybody else, that adding females was just going to complicate our experiments,” Levy added, “And so we said, ‘why not test this.’”

Testing assumptions

The researchers studied genetically identical males and females from a common strain of lab mouse in a circular open field — a standard lab setup for behavioral neuroscience experiments. In practice, the test involved placing a mouse in a 5-gallon Home Depot bucket for 20 minutes and using a camera to record the mouse’s movements and behaviors in 3D as it freely explored the space. The researchers swabbed each female mouse to determine its estrous status and repeated the bucket test with the same individual multiple times.

“This is a very interesting example of how assumptions that affect the way that we conduct and design our science are sometimes just assumptions — and it is important to directly test them, because sometimes they’re not true.” Dana Levy, a research fellow at Harvard Medical School

The team analyzed the videos with MoSeq, an artificial intelligence technology previously developed by the lab. The technology uses machine learning algorithms to break down a mouse’s movements into around 50 different “syllables,” or components of body language: short, single motions such as rearing up, pausing, stepping, or turning. With MoSeq, the researchers gathered in-depth, high-resolution data about the structure and pattern of mouse behavior during each session.

The researchers found that estrous status had very little effect on exploratory behavior in female mice. Instead, patterns of behavior tended to vary much more across female mice than they did throughout the estrous cycle.

“If you give me any random video from our pile, I can tell you which mouse it is. That’s how individualized the pattern of behavior is,” Datta said, which suggests that in behavioral studies, “a dominant aspect of variation in the data is the fact that individuals have subtly different life histories.”

When the researchers compared female and male mice, they found something that surprised them: Males also exhibited individuality of behavior, but they had more behavioral variation within a single mouse and between mice than females.

“People have been making this assumption that we can use male mice to reliably make comparisons within and across experiments, but our data suggest that female mice are more stable in terms of behavior despite the fact that they have the estrous cycle,” Datta said.

A case for change

Scientists generally agree that including female mice is important from a fairness perspective, Datta noted, yet some have remained concerned that it could complicate their research. For him, the new findings make a strong scientific case for using female mice in experiments.

“The fact that female behavior is more reliable suggests that including females might actually decrease the overall variability in your data under many circumstances,” Datta said.

Based on their findings, researchers in the Datta lab have already switched from male mice to mixed groups or female mice in their other experiments that involve circular, open-field testing.

Datta cautioned that the study looks at only one mouse strain in one lab setup, and so the results cannot be generalized to other strains and setups without further testing. However, he noted that the strain and setup are commonly used in neuroscience research, including in early-stage drug development to test how a potential drug affects mouse locomotion.

Datta said that the findings “should encourage folks who are interested in drug development in this context to include both sexes in their analysis.”

Now, Datta and Levy are interested in exploring how internal states beyond hormonal status, such as hunger, thirst, pain, and illness, affect exploratory behavior in mice.

“The question is, who wins in this tug-of-war between your current internal state and your individual identity,” Levy explained.

They also want to delve deeper into the neural basis of the individuality of mouse behavior that they saw in the study.

“I was shocked by how much stable variation between individuals we were observing — it’s like these mice really are individuals,” Datta said. “We’re used to thinking of lab mice as interchangeable widgets, but they’re not at all. So, what is controlling these individualized patterns of behavior?”

“We want to understand the mechanisms of individuality: how variability between individuals comes about, how it affects behavior, what can alter it, and what brain regions support it,” Levy added.

To this end, the Datta lab is examining mouse behavior from birth until death to understand how individualized patterns of behavior emerge and crystallize during development, and how they change throughout life.

The researchers also hope that their work will open the door for more rigorous, quantitative research on whether and how the estrous cycle affects mouse behavior in other contexts, such as completing complex tasks.

“This is a very interesting example of how assumptions that affect the way that we conduct and design our science are sometimes just assumptions — and it is important to directly test them, because sometimes they’re not true,” Levy said.

Additional authors include Nigel Hunter, Sherry Lin, Emma Robinson, Winthrop Gillis, Eli Conlin, and Rockwell Anyoha of HMS.

Datta is on the scientific advisory boards of Neumora, Inc., and Gilgamesh Pharmaceuticals, which have licensed the MoSeq technology.

The research was supported by the NIH (U19NS113201; RF1AG073625; R01NS114020), the Brain Research Foundation, the Simons Collaboration on the Global Brain, the Simons Collaboration for Plasticity in the Aging Brain, the Human Frontier Science Program, and the Zuckerman STEM Leadership Program.

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Major New Study Reveals New Similarities and Differences Between Mice and Humans

Credit: Ryan Lister, University of West Australia

Powerful clues have been discovered about why the human immune system, metabolism, stress response, and other life functions are so different from those of the mouse. A new, comprehensive study of the mouse genome by an international group of researchers including Penn State University scientists reveals striking similarities and differences with the human genome. The study may lead to better use of mouse models in medical research.

The findings are reported by the Mouse ENCODE Consortium online on November 19, 2014 and in print on November 20 in the study's main paper in Nature and in several other recent and future publications. They examine the genetic and biochemical programs involved in regulating mouse and human genomes. Ross Hardison , the director of the Huck Institute for Comparative Genomics and Bioinformatics at Penn State University, is the senior corresponding author or co-senior author for four of the five new papers by the consortium, including the paper in Nature.

"We didn't know before these research results that there are a large number of genes with expression levels systematically different between mouse and human," Hardison said. The results offer insights into how gene regulation impacts systems important to the biology of humans and other mammals. The results also provide new information to determine how best to use the mouse as a model for studying human biology and disease, and may help to explain some of the limitations of using the mouse for specific kinds of studies.

"Now we also know which genes have expression patterns that are shared between mouse and humans," Hardison said. "For biological processes using genes with conserved expression patterns, the mouse is an excellent model for certain aspects of human biology."

The scientists also found that, in general, the systems that are used to control gene activity have many similarities in mice and humans, and that the basic structure of these systems has been conserved in both species throughout evolutionary time. The researchers found that differences appeared for specific genes and regulatory elements. "Gene regulation is an equation with many possible solutions," said John Stamatoyannopoulos at the University of Washington, a co-senior author with Hardison of the main Nature paper.

The Mouse ENCODE (ENCyclopedia Of DNA Elements) project is building a comprehensive catalog of functional elements in the mouse genome, and is comparing them to those in the human genome. Such elements include genes that code for proteins, non-protein-coding genes, and regulatory elements that control which genes are turned on or off, and when they are turned on or off. Bing Ren at the University of California, San Diego, also a co-senior author with Hardison of the Nature study, said "This is the first systematic comparison of the mouse and human at the genomic level."

The portion of the Mouse ENCODE effort centered at Penn State focused on comparing mouse and human gene expression and regulatory elements during cell differentiation. This work was done in collaboration with Yu Zhang , associate professor of statistics at Penn State, Feng Yue , assistant professor of biochemistry and molecular biology at Penn State's College of Medicine, and other researchers. "Comparison of the regulatory landscape between mouse and human reveals complex relationships, with some regulatory regions being strictly conserved between mouse and human, other regulatory regions being lost or acquired along each evolutionary lineage -- perhaps reflecting adaptation to different environments, and other regulatory regions being re-used in different tissues," Hardison said. "One would expect that the strictly conserved regulatory regions were particularly important, and this is true, but our collaborative studies have revealed an unexpected basis for their importance." The Mouse ENCODE work revealed that these strictly conserved regulatory regions are active across different tissues, including blood, heart, brain, and others, to a much greater extent than had been previously appreciated. The multiple functions of these regulatory regions may explain the stronger selective pressure during evolution, thus leading to their strict conservation.

The broad, global approaches used in Mouse ENCODE allow investigators to see which genes are expressed in similar patterns and levels between mouse and human, and which ones have divergent patterns. "This information from Mouse ENCODE will enable investigators to make data-driven interpretations of the rich body of information in mouse model systems for potential translation to insights about human biology and health," Hardison said. "For genes with conserved expression patterns, the translation may be quite direct, whereas for others the divergence in expression patterns needs to be incorporated into the inferences for human biology."

More than a dozen companion studies have appeared or will appear in journals including Nature, Nature Communications, Genome Research, and Genome Biology. ENCODE data are freely shared with the biomedical community, and the mouse resource has already been used by researchers outside of ENCODE in about 50 publications.

ENCODE was started with funds from the American Recovery and Reinvestment Act of 2009 and now is supported by the National Human Genome Research Institute (NHGRI), part of National Institutes of Health (NIH). "The mouse has long been a mainstay of biological research models," said NHGRI Director Eric Green , M.D., Ph.D. "These results provide a wealth of information about how the mouse genome works, and a foundation on which scientists can build to further understand both mouse and human biology. The collection of Mouse ENCODE data is a tremendously useful resource for the research community."

Members of Hardison's lab team who were involved in the Mouse ENCODE research are Graduate Students Swathi Ashok Kumar (Genetics), Christapher Morrissey (Bioinformatics and Genomics), Deepti Jain (Biochemistry, Microbiology, and Molecular Biology), Nergiz Dogan (Biochemistry, Microbiology, and Molecular Biology), Marta Byrska-Bishop (Molecular, Cellular, and Integrative Biosciences), and Kuan-Bei Chen (Computer Science and Engineering); Postdoctoral Fellow Weisheng Wu , Ph.D; Project Manager and Senior Research Associate Cheryl A. Keller , Ph.D; Programmer/Analysts Belinda Giardine and Robert Harris , Ph.D; and Laboratory Assistant Maria Long .

Ross Hardison: [email protected] , 814-863-0113 Barbara Kennedy (PIO): [email protected] , 814-863-4682

Why Do Medical Researchers Use Mice?

From formulating new cancer drugs to testing dietary supplements, mice and rats play a critical role in developing new medical wonders. In fact, 95 percent of all lab animals are mice and rats, according to the Foundation for Biomedical Research (FBR).

Scientists and researchers rely on mice and rats for several reasons. One is convenience: rodents are small, easily housed and maintained, and adapt well to new surroundings. They also reproduce quickly and have a short lifespan of two to three years, so several generations of mice can be observed in a relatively short period of time.

Mice and rats are also relatively inexpensive and can be bought in large quantities from commercial producers that breed rodents specifically for research. The rodents are also generally mild-tempered and docile, making them easy for researchers to handle, although some types of mice and rats can be more difficult to restrain than others . [ Why Do Mice Poop So Much? ]

Most of the mice and rats used in medical trials are inbred so that, other than sex differences, they are almost identical genetically. This helps make the results of medical trials more uniform, according to the National Human Genome Research Institute. As a minimum requirement, mice used in experiments must be of the same purebred species.

Another reason rodents are used as models in medical testing is that their genetic, biological and behavior characteristics closely resemble those of humans, and many symptoms of human conditions can be replicated in mice and rats. "Rats and mice are mammals that share many processes with humans and are appropriate for use to answer many research questions," said Jenny Haliski, a representative for the National Institutes of Health (NIH) Office of Laboratory Animal Welfare.

Over the last two decades, those similarities have become even stronger. Scientists can now breed genetically-altered mice called "transgenic mice" that carry genes that are similar to those that cause human diseases. Likewise, select genes can be turned off or made inactive, creating "knockout mice," which can be used to evaluate the effects of cancer-causing chemicals (carcinogens) and assess drug safety, according to the FBR.

Rodents also make efficient research animals because their anatomy, physiology and genetics are well-understood by researchers, making it easier to tell what changes in the mice's behaviors or characteristics are caused by.

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Some rodents, called SCID (severe combined immune deficiency) mice, are naturally born without immune systems and can therefore serve as models for normal and malignant human tissue research , according to the FBR.

Some examples of human disorders and diseases for which mice and rats are used as models include:

Hypertension
Respiratory problems
Parkinson's disease
Alzheimer's disease
Cystic fibrosis
HIV and AIDs
Heart disease
Muscular dystrophy
Spinal cord injuries

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Mice are also used in behavioral, sensory, aging, nutrition and genetic studies, as well as testing anti-craving medication that could potentially end drug addiction .

"Using animals in research is critical to scientific understanding of biomedical systems leading to useful drugs, therapies and cures," Haliski told Life's Little Mysteries.

Originally published on Live Science .

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Drugs That Work In Mice Often Fail When Tried In People

Richard Harris

Most new drugs don't work when tested in people. One of the big reasons is the use of animals in research.

Most potential new drugs fail when they're tested in people. These failures are not only a major disappointment, they sharply drive up the cost of developing new drugs.

A major reason for these failures is that most new drugs are first tested out in mice, rats or other animals. Often those animal studies show great promise.

But mice aren't simply furry little people, so these studies often lead science astray. Some scientists are now rethinking animal studies to make them more effective for human health.

When scientists first started using animals in research over a century ago, the animals were not regarded as human stand-ins. Scientists studying rats were initially trying to understand rats, says Todd Preuss , an anthropologist at the Yerkes National Primate Research Center at Emory University.

"As this process went on, people stopped seeing them as specialized animals and started seeing them more and more as prototypical mammals," Preuss says.

But is a rat really a generic mammal? Preuss says emphatically no. But that's how rodents were pitched when they became products sold to scientists.

"It wasn't strictly a financial interest," he says. The sellers "really believed that you could do almost anything" with these animals. "You could learn about almost any feature of human organization, you could cure almost any disease by studying these animals."

That was a dangerous assumption. Rats and humans have been on their own evolutionary paths for tens of millions of years. We've developed our own unique features, and so have the rodents.

So it should come as no surprise that a drug that works in a mouse often doesn't work in a person. Even so, Preuss says there's tremendous momentum to keep using animals as human substitutes. Entire scientific communities are built up around rats, mice and other lab animals.

"Once these communities exist, then you have an infrastructure of knowledge: how to raise the animals, how to keep them healthy," Preuss says. "You have companies that spring up to provide you with specialized equipment to study these animals."

The rat holding facility at Hazelton Laboratories in Washington, D.C., in 1967. Fox Photos/Getty Images hide caption

The rat holding facility at Hazelton Laboratories in Washington, D.C., in 1967.

Chances are, people studying the same disease study the same tailor-made strain of animal. Journals and funding agencies actually expect it.

"So there's a whole institution that develops," Preuss says.

And it's hard to interrupt that culture. (Preuss spoke about this subject in a 2016 talk at the National Institutes of Health.)

You can get a glimpse of the scale of this enterprise by passing through one of hundreds of facilities nationwide devoted to the care and feeding of mice. On the Stanford University campus, attendants roll supply carts through fluorescent-lit hallways and past row after row of doors at an expansive mouse facility.

I'm guided through the labyrinth by Joseph Garner , a behavioral scientist at the Stanford University Medical Center. We go into a windowless room stacked floor to ceiling with seemingly identical plastic cages full of mice.

The philosophy behind mouse research has been to make everything as uniform as possible, so results from one facility would be the same as the identical experiment elsewhere.

But despite extensive efforts to be consistent, this setup hides a huge amount of variation. Bedding may differ from one facility to the next. So might the diet. Mice respond strongly to individual human handlers. Mice also react differently depending on whether their cage is up near the fluorescent lights or hidden down in the shadows.

Garner and colleagues tried to run identical experiments in six different mouse facilities, scattered throughout research centers in Europe. Even using genetically identical mice of the same age, the results varied all over the map.

Garner says scientists shouldn't even be trying to do experiments this way.

"Imagine you were doing a human drug trial and you said to the FDA, 'OK, I'm going to do this trial in 43-year-old white males in one small town in California,'" Garner says — a town where everyone lives in identical ranch homes, with the same monotonous diets and the same thermostat set to the same temperature.

"Which is too cold, and they can't change it," he goes on. "And oh, they all have the same grandfather!"

The FDA would laugh that off as an insane setup, Garner says.

"But that's exactly what we do in animals. We try to control everything we can possibly think of, and as a result we learn absolutely nothing."

Garner argues that research based on mice would be more reliable if it were set up more like experiments in humans — recognizing that variation is inevitable, and designing to embrace it rather than ignore it. He and his colleagues have recently published a manifesto , urging colleagues in the field to look at animals in this new light.

"Maybe we need to stop thinking of animals as these little furry test tubes that can be or even should be controlled," he says. "And maybe instead we should think of them as patients."

That could solve some of the problems with animal research, but by no means all.

Scientists make far too many assumptions about the underlying biology of disease when creating animal models of those illnesses, says Gregory Petsko , who studies Alzheimer's disease and other neurological disorders at the Weill Cornell Medical School.

"It's probably only when you get to try your treatments in people that you're really going to have any idea how right those assumptions were," Petsko says.

In his field, the assumptions are often poor, or downright misleading. And Petsko says this mindset has been counterproductive. Scientists in his field have "been led astray for many years by relying so heavily on animal models," he says.

For many years that was simply the best that science could do, Petsko says. So he doesn't fault his colleagues for trying.

"What I am saying is at some point you have to cut your losses. You have to say, 'OK, this took us as far as it could take us, quite some time ago.'"

For neurological diseases, Petsko says, scientists might learn more from studying human cells than whole animals. Animals are still useful for studying the safety of potential new treatments, but beyond that, he says, don't count on them.

Preuss at Emory agrees that using animals as models of disease is a big reason that many results in biomedical research aren't readily reproducible. "I think that we have means to resolve that, though."

How? "You have to think outside of the model box," he says. Mice and rats aren't simplified humans. Scientists should stop thinking they are.

But Preuss says scientists can still learn a lot about biology and disease by studying animals — for example, by comparing how humans and other animals differ, or where they share common traits. Those can reveal a lot about biology without assuming that what's true in a rat is likely true in a human.

"Scientists need to break out of a culture that is hampering progress," Preuss says. That's tough to do right now, in a world where science funding is on the chopping block. Many scientists are reluctant to take a risk that could backfire. But the upside could benefit us all, in the form of a better understanding of disease, and effective new drugs.

Richard Harris did some of the reporting for this story while researching his book Rigor Mortis: How Sloppy Science Creates Worthless Cures, Crushes Hope, and Wastes Billions. You can contact him at [email protected] .

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Molecule restores cognition and memory in Alzheimer’s disease mouse study

Person in lab coat, surgical mask and gloves holds a film up to a light and looks at images of brain scans on it

In a new study, a molecule identified and synthesized by UCLA Health researchers was shown to restore cognitive functions in mice with symptoms of Alzheimer’s disease by effectively jump-starting the brain’s memory circuitry.

If proven to have similar effects in humans, the candidate compound would be novel among Alzheimer’s disease treatments in its ability to revitalize memory and cognition, study authors said.

“There is really nothing like this on the market or experimentally that has been shown to do this,” said study lead author Dr. Istvan Mody, the Tony Coelho Professor of Neurology and distinguished professor of physiology at UCLA Health.

The molecule, DDL-920, works differently from recent FDA-approved drugs for Alzheimer’s disease such as lecanemab and aducanumab, which remove harmful plaque that accumulates in the brains of Alzheimer’s disease patients. While removing this plaque has been shown to slow the rate of cognitive decline, it does not restore the memory or remedy cognitive impairments.

“They leave behind a brain that is maybe plaqueless, but all the pathological alterations in the circuits and the mechanisms in the neurons are not corrected,” Mody said.

In the study, published in the journal The Proceedings of the National Academy of Sciences, UCLA researchers led by Dr. Istvan Mody and Dr. Varghese John, a professor of neurology and director of the Drug Discovery Laboratory (DDL) at the Mary S. Easton Center for Alzheimer's Disease Research and Care at UCLA sought to find a compound that could figuratively flip the switch back on in the brain’s memory circuitry.

Similar to a traffic signal, the brain fires off electric signals at different rhythms to start and stop various functions. Gamma oscillations are some of the highest-frequency rhythms and have been shown to orchestrate brain circuits underlying cognitive processes and working memory – the type of memory used to remember a phone number. Patients with early Alzheimer’s disease symptoms such as mild cognitive impairment have been shown to have reduced gamma oscillations, Mody said.

Other studies attempted to use neuromodulation techniques to stimulate gamma oscillations to restore memory. Auditory, visual or transcranial magnetic stimulation at a frequency of 40 hertz – similar to the frequency of a cat’s purr – worked to dissolve plaques in the brain but again did not show notable cognitive enhancements, Mody said.

In this latest study, Mody and his team sought to tackle the problem from a different perspective. If they could not jump-start these memory circuits using external tools, perhaps there was a way to trigger these electrical rhythms from the inside using a molecule.

Specifically, they needed a compound to target certain fast-firing neurons, known as the parvalbumin interneurons, that are critical in generating gamma oscillations and therefore memory and cognitive functions. However, certain chemical receptors in these neurons that respond to the chemical messenger known as Gamma-aminobutyric acid, or GABA, work like brake pedals to reduce the gamma oscillations entrained by these neurons.

Mody, John and their team identified the compound DDL-920 to antagonize these receptors, allowing the neurons to sustain more powerful gamma oscillations.

To test whether this would result in improved memory and cognition, researchers used mice that were genetically modified to have symptoms of Alzheimer’s disease.

Both these Alzheimer’s disease-model mice and wild-type mice underwent baseline cognitive testing in a Barnes maze – a circular platform surrounded by visual clues and containing one escape hole. The maze is used to measure how well rodents can learn and remember the location of the escape hole.

After the initial tests, researchers orally administered DDL-920 to the Alzheimer’s model mice twice daily for two weeks. Following treatment, the Alzheimer’s disease-model mice were able to recall the escape hole in the maze at similar rates as the wild-type mice. Additionally, the treated mice did not display any abnormal behavior, hyperactivity or other visible side effects over the two-week period.

Mody said that while the treatment was effective in mice, much more work would be needed to determine if the treatment would be safe and effective in humans. Should it ultimately prove to be effective, the drug could have implications for treatments of other diseases and health conditions that have diminished gamma oscillations, such as depression, schizophrenia and autism spectrum disorder, Mody said.

“We are very enthusiastic about that because of the novelty and the mechanism of action that has not been tackled in the past,” Mody said.

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Health Evaluation of Experimental Laboratory Mice

Good science and good animal care go hand in hand. A sick or distressed animal does not produce the reliable results that a healthy and unstressed animal produces. This unit describes the essentials of assessing mouse health, colony health surveillance, common conditions, and determination of appropriate endpoints. Understanding the health and well-being of the mice used in research enables the investigator to optimize research results and animal care.

INTRODUCTION

Both investigative and veterinary staffs monitor the health and well-being of mice that are used in research. Indeed, this level of responsibility and care is mandated by the Public Health Service based on the Guide for the Care and Use of Laboratory Animals ( National Research Council. 2011 ). The Guide is “intended to assist investigators in fulfilling their obligation to plan and conduct animal experiments in accord with the highest scientific, humane, and ethical principles.” All investigators should become familiar with the Guide ( http://grants.nih.gov/grants/olaw/Guide-for-the-Care-and-Use-of-Laboratory-Animals.pdf ).

Careful observation of mice in their home cage can provide a wealth of information about the health and welfare of the animals. Activity, nest building, interaction with cage mates and general appearance are indicators of general health and well-being. Hands-on physical examination provides an assessment of the animal’s hydration, body condition, observable abnormalities, and the presence of palpable anomalies in the abdomen. We will outline details that will be not only useful to the investigator, but quick and easy to perform as well.

Several infectious agents have been identified over the years that have either adverse affects on animal health and/or research outcome. For this reason, most mouse colonies are maintained as Specific Pathogen Free (SPF), free of defined infectious agents. SPF status is generally monitored by exposing sentinel animals to dirty bedding of colony animals and testing the sentinel animals. Tremendous effort and expense is expended maintaining SPF colonies and it is critical that all staff entering colonies and handling animals understand disease transmission and the importance of good practices to prevent pathogens from being introduced or spread.

Laboratory mice develop a number of common clinical conditions which will be described, together with recommended treatments and suggested endpoints. For instance, dermatitis is a poorly understood condition that is common and problematic, especially in animals with a C57BL/6 background. Some congenital conditions such as hydrocephalus or microphthalmia are also seen with a C57BL/6 background. Behavioral differences exist between strains and lines of animals with some prone to fighting when males are co-housed. BALB/c, FVB and SJL strains are particularly troublesome in this regard although management practices can help reduce fighting.

The Guide is based on three principles: Replace, Reduce and Refine. Replace animals whenever possible, reduce the number to lowest possible that will produce accurate conclusions, and refine the experimental paradigm to improve the science and the care of the animal. These principles were introduced by British investigators William Russell and Rex Burch in 1959 in response to the moral and ethical concerns associated with the use of animals in research ( Russell and Burch, 1959 ; Flexnell, 2002). Refinement is a principle that directly relates to the topic at hand, and can be as simple as adding palatable food on the cage floor or as sophisticated as utilizing telemetry to monitor physiology and activity.

The 8 th Guide published in 2011 also emphasizes the refinement of end points and states that “The use of humane endpoints contributes to refinement by providing an alternative to endpoints that result in unrelieved or severe animal pain and distress.” In other words, an animal should be euthanized at the earliest possible point that will provide experimental data in order to minimize suffering. A variety of refined endpoints for multiple species have been published and they include data-based systems for assessing animals, drops in core body temperature as an alternative endpoint, endpoints for tumors and ascites production, and changing from an awake sepsis model to an anesthetized sepsis model, among others. ( Hendriksen and Steen, 2000 ; Morton, 2000 ; Olfert and Godson, 2000 ; Minecci et al., 2007 ; Paster et al., 2009 ; Sass, 2000 ; Stokes, 2000 ; Stokes, 2002 ; Toth, 1997 ; Toth, 2000 ; Toth and Gardiner, 2000 ; Wallace, 2000 ).We discuss endpoints in mice for the consideration of the investigator.

COLONY HEALTH SURVEILLANCE

Colony health surveillance is typically part of the overall veterinary care program to ensure the SPF status of the animal facility. Many microbial outbreaks are subclinical in mice. Therefore, microbiological surveillance of colonies is required for detection and to ensure the appropriate health status of the colony and individual mouse. Sentinels are animals that are free of excluded microorganisms and are exposed to dirty bedding of colony animals to determine if excluded microorganisms are present in the colony animals. Health surveillance testing of sentinel and / or colony (non-sentinel) animals can include the following: gross, histopathology, and parasitology assessments, testing of serology samples for antibodies or antigens, culture or isolation of microorganisms, and molecular diagnostics such as the polymerase chain reaction (PCR) test. The diagnostic procedures utilizing samples collected from live animals for colony health surveillance include those used to assess the health of individual mice. For example, collection of: a fur sample for an ectoparasitology exam, fecal material for endoparasitology and PCR testing, and blood for serology testing can help determine the health status of an individual animal. Additional details regarding surveillance programs can be found in the text, Fox, et al., 2002 . “Laboratory Animal Medicine,” 2 nd ed. An example of commercial laboratory services for health surveillance can be found at: http://www.criver.com/enUS/ProdServ/ByType/ResAnimalDiag/Pages/home2.aspx .

Enormous expense is involved with maintaining SPF colonies. The protective personal equipment (PPE) used and the regular purchase and testing of sentinel animals is a significant investment by the facility/institution. Outbreaks of disease agents exponentially increase that cost. To contain the disease outbreak extensive additional testing is necessitated. For this reason facility staff and investigative staff should do their utmost to follow facility procedures, traffic patterns and standard operating procedures. For instance, in a facility where animals are changed in hoods using appropriate micro-isolator techniques then investigative staff should follow those same practices (e.g., not open cages outside of the hood). Another important consideration in this same vein is following appropriate procedures for introducing animals from outside facilities. Typically, there is a gatekeeper through who all imports of animals are managed. This is to ensure that their health status is known prior to arrival and confirmed after arrival before introduction into the main animal colony.

EXAMINATION AND ASSESSMENT OF THE MOUSE

An overall assessment of the health and welfare of a research mouse includes an evaluation of the animal in its home cage and a hands-on exam. Because mice are easily stressed by handling, the cage side exam should be performed first. Observing the mouse in its home cage will provide information about the animal’s overall appearance and activity level, the interaction with the environment, including nest building, and its behavior with respect to its cage mates. The hands-on examination allows assessment of observable abnormalities, hydration status, body condition, and the presence of abnormalities in the bones, genitals and abdomen.

Home cage evaluation

Mice are inquisitive and active and will generally be observed moving around the cage, grooming, eating, drinking and interacting with cage mates; particularly after being stimulated by having their cage picked up and moved from the shelf or rack.

Behavioral indicators of a welfare issue can be obvious including wounds and limping, hunched posture, dull or sluggish movements, large or open tumors, or a mouse that does not move when the cage is manipulated. Many behaviors are more subtle and non specific and will take practice and time to evaluate. Among these is nest building. All mice will build a nest if given suitable material ( Hess et al., 2008 ). A mouse that is placed in a new cage with nesting material and has not built a nest by the next day warrants veterinary attention and possibly euthanasia. Similarly, the absence of feces in a cage after the mouse has been housed several hours suggests the animal has not been eating.

Mice are prey species and will generally mask signs of pain. If evaluating mice for pain after surgery or a procedure that has the potential to be painful, the observer needs to be quiet and monitor the mice without moving the home cage. Mice, like all animals express emotion including pain through facial expressions. Many of the expressions are identical to those expressed by humans in pain including squinting the eyes and contracting the skin around the nose and mouth. Mice may also pull their ears back ( Langford et al., 2010 ; Figure 1 ). Mice in pain are less active than normal, although this type of qualitative assessment is difficult to evaluate on cage side exam unless it is severe ( Clark et al., 2004 ; Roughan et al., 2009 ).

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Facial expressions in mice indicating pain and/or distress include squinted eyes, contracted skin around nose and ears pulled back.

Developing and assigning a study specific pain scale can be an effective tool to facilitate communication between observers and can serve as a convenient indication when an animal’s condition is deteriorating or has reached a clinical end point. Table 1 is an example of a general clinical pain score.

Assessing Pain and Distress in Mice

PAIN AND DISTRESS ASSESSMENT	EXAMPLES
1) no indication of pain and distress	Normal; well groomed; alert; active; good condition; asleep or calm; normal appetite; BCS=3,4 or 5
2) mild or anticipated pain and distress	Not well groomed; awkward gait; slightly hunched; looks at wound or pulls away when area touched; mildly agitated; BCS=2
3) moderate pain and distress	Rough hair coat; dirty incision; squinted eyes; moves slowly; walks hunched and/or slowly; depressed or moderately agitated; slight dehydration; pruritic; restless; uncomfortable; not eating or drinking; BCS= 2-.
*4) severe pain and distress	Very rough hair coat; eyes sunken (severe dehydration); slow to move or non-responsive when coaxed; hunched; large abdominal mass; dyspnea; self mutilating; violent reaction to stimuli or when approached; BCS=1

Handling and Hands on evaluation

Laboratory mice are generally docile but will move quickly or jump away from the person trying to restrain them, and some strains may bite. Mice can be moved a short distance for examination by being picked up at the base of the tail and placed onto the top of their cage allowing the mouse to grasp the bars with the forefeet and direct its effort away from the handler. Very obese or pregnant mice should have a hand placed under the abdomen to prevent the heavy abdominal contents from compressing the diaphragm and limiting respiration. Retain control of the tail to prevent the mouse from escaping and potentially harming itself.

Restraint . With the animal restrained on top of the cage, run a finger over the animal’s coat to feel for wounds that may be covered with fur and feel for masses that may not have been obvious when the mouse was moving around the cage.
Hydration. Severely dehydrated mice will be weak and often will look paralyzed in their rear legs. These mice may also have trouble gripping the cage bars with their forefeet. Other symptoms of severe dehydration include sunken or recessed eyes and fuzzy facial fur, which results due to piloerection. More moderate dehydration can be detected by pinching the skin over the shoulder blades. In a well hydrated mouse the skin will quickly return to its original shape. The skin remaining bunched up is an indication of dehydration.

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Body Condition Scoring (BCS) is a quick, easy and reliable method for assessing mouse health. It utilizes a scoring system of 1 to 5 with 3 being the optimal condition, 1 being emaciated and 5 being obese.

Restraint and physical examination

While holding the animal by the tail with the little finger, use the forefinger and thumb to gently pin down the head and grasp the loose skin over the back of the neck. The remaining fingers can be used to scruff the skin along the back of the mouse, allowing the mouse to be picked up and restrained. Once restrained, examine the animal from its nose to the tail. Perform the exam in the same order each time an animal is examined to establish a pattern that will help prevent overlooking a more subtle abnormality.

Face and mouth. Evaluate the eyes, ears, face and neck for abnormalities. Examine the incisors by gently moving the lips using a cotton tipped applicator. Evaluate the color of the mucous membranes, which should be pink. The mucous membranes around the eyes and the skin of albinos appearing pale, bluish, or brick red in coloration indicate conditions such as anemia, hypoxia, or systemic infection and circulatory failure. These are near terminal conditions and indications for euthanasia. Note that a greenish hue may be due to genetic constructs with green fluorescent protein and are expected in some strains.
Feet and limbs. Next, examine the feet and limbs. Explore any gait abnormalities observed during the cage side exam. Lameness in one limb may be due to a wound on the foot or bony tumor that can be felt along the length of the bone. Pinch the toes to distinguish between weakness in the limbs and paralysis. Weak mice will generally pull away when the toes are pinched.
Genital abnormalities . Evaluate the female mammary chain for masses, abnormalities around the nipples or irregularities around the vulva. Examine the male penis by gently sliding back the prepuce. A purple or distended penis during exam likely indicates a urinary obstruction, which is a painful and life threatening condition. Check the rectal area for swelling, trauma or prolapsed tissue. Swellings adjacent to the rectum in older male mice likely indicate cysts in the reproductive glands and are generally benign.
Abdominal palpation . Palpate the abdomen by gently compressing the contents between the fingers from just under the ribs down to the hips. Common abnormalities palpated in the cranial aspect of the abdomen include tumors of the liver and spleen. In male mice, the most common abnormality felt in the middle part of the abdomen is due to enlargements of the kidneys secondary to urinary obstruction. In pregnant females, the distended uterus can be palpated in the mid abdomen and can extend up under the rib cage. In older females or retired breeders, masses in the mid abdomen are generally due to uterine tumors. The bladder can be felt in the caudal abdomen. Mice will generally urinate as a stress response when picked up. A large or distended bladder can indicate an obstruction. In males, the glands of the reproductive organs can become enlarged and distended with fluid as the mice age which can be palpated in the caudal abdomen.

See http://www.ors.od.nih.gov/sr/dvr/od/Documents/Post_Op_Form.docx

COMMON CLINICAL HEALTH CONDITIONS OF MICE

While mice are valuable research tools, they are also living animals with characteristics and health issues that may influence or be of concern while conducting a study. A holistic approach to evaluation of clinical conditions, including influences on the research and the welfare of the animals will allow the investigative and veterinary teams to make appropriate decisions. The discussion here will cover the most frequently seen conditions, but is not an exhaustive listing. The photographs included will illustrate many of these conditions. Treatment suggestions are based on the author’s clinical experience but should always be discussed with the veterinary team at your institution. Also, keep in mind that the physiology of the animal may be affected by responses to the condition itself and by any treatments administered. Finally, euthanasia may be the most appropriate response for both humane and research related reasons.

Skin lesions

Skin lesions are one of the most commonly observed clinical problems in mice and arise from many different causes. Often these can be clinically managed to keep the mouse on study or retain it for breeding, but strain predilections must also be considered. The immune system of the mouse will be activated to fight these lesions, which must be considered in light of the study protocol.

Fight wounds : Most commonly seen in co-housed male mice, especially from strains such as BALB/c, SJL, and FVB, however this behavior can also occur between females or in mixed sex groups. The typical presentation is a cluster of wounds on the rump, hips, and/or genital region, which may extend to the trunk of the body or forelegs. Often there is one aggressor in the cage which can be removed while beginning treatment of the other mice with antibiotics and analgesics systemically and/or topically. If the wounds are not severe, mice generally heal well. If wounds are severe, humane endpoint criteria should be discussed with your veterinary staff. Fighting can be minimized by housing only littermate males together, or single housing particularly aggressive strains.
Ear dermatitis : Often related to ear tags used for identification. Although tags are normally very well accepted by the animals, problems may occur due to tag placement, sensitivity to the tag metals, or secondary to fighting. If irritation is noted, it is advisable to remove the tag if still present, and to use antibiotic and analgesic therapies to treat secondary infection and pruritis. Clipping of the hind toenails helps reduce trauma due to scratching.
Alopecia : Especially when seen in patches around the face or in one location on several mice within a group, is a sign of what is termed “barbering”. The skin is generally healthy, and short stubby hairs may be seen in the alopecic area. In some cases the whiskers or eyelashes may be missing. This condition is caused by over grooming by an animal’s cage mates or itself. It may be a dominance action or an obsessive compulsive grooming disorder. Unless there is secondary ulceration or inflammation of the skin, no medical treatment is necessary, but increased environmental enrichment has potential benefit.
Dermatitis : Includes ulcerative dermatitis (common in C57BL/6 background mice), miliary dermatitis, muzzle dermatitis involving the hair follicles, (referred to as furunculosis or botryomycosis), and contact dermatitis. Erosions of the skin or small raised scabbed lesions may be seen. If lesions are deep, large, or bleeding then aggressive care with analgesics, anti-inflammatory medications, and antibiotics, or euthanasia are indicated. For less severe lesions, control of infection by skin bacteria relieves the irritation and may allow healing; treatment may be systemic or topical. Many different treatment regimens have been used for ulcerative dermatitis including nutritional supplements and NK1 receptor antagonists (Lawson, G.W., et al. 2011).
Hyperkeratosis : Thickening of the skin without shedding of the surface dead epithelium is often a sign of irritation. It may also be a sign of neoplasia, bacterial skin infection, or mite infestation. Skin scrapings, fur plucks, or biopsy may be used diagnostically to guide treatment decisions. In nude mice Corynebacterium bovis has been noted to cause a flaky white dandruff-like skin condition which may be responsive to antibiotic treatment. Sterile caging is beneficial to reduce incidence.
Otitis : Head tilt or circling behavior can be associated with inflammation within the inner or outer ear. External lesions often respond to topical antibiotic, while inner ear conditions with nerve damage are more complicated and may lead to indications for euthanasia if the animal cannot maneuver to reach feed and water.
Tail lesions : May present as dermatitis, as fight wounds, or as concentric rings with hyperkeratosis. The latter has historically been related to very low humidity and is known as “ringtail”. Granulomas may form after tail biopsies for genotyping of mice. Treatments are the same as similar lesions found on the body. Small nicks used for collection of blood from the tail vein(s) will scab and generally heal without medical treatment.

Lumps and Bumps

“Lumps and Bumps” require further diagnostic evaluation to determine the cause and significance. Common causes are tumors (spontaneous or study induced), abscesses, cysts, lymphadenopathy, salivary gland hyperplasia, and reactions to injections especially if adjuvants are used.

Mammary tumors : A common problem in female mice which may be observed almost anywhere on the trunk of the body due to the extensive distribution of mammary tissue. These are subcutaneous, may be smooth or rough, and are usually easily moveable under the skin. In mice these tend to be malignant. Treatment is not advised.
Tumors : These should be evaluated with consideration of humane endpoints. Their size, location, secondary effects, etc. will be considered in decisions to observe, surgically remove, or recommend euthanasia.
Abscesses : Can occur in any location, but the most common are related to bite wounds, necrotic tumors, or blocked ducts to normal exocrine glands such as the preputial glands of male mice. They are usually a soft to firm swelling which may or may not be inflamed. Needle aspirate allows drainage of pus or caseous exudate which can be submitted for culture and antibiotic sensitivity testing to guide treatment plans. In some cases the abscess ruptures at the skin surface spontaneously. Treatments include draining and/or flushing the abscess, antibiotic therapy, and clean soft bedding if located ventrally. If not responsive to therapy, euthanasia must be considered.
Lymphadenopathy : Usually noted under the legs or in the neck region, but may occur anywhere lymph nodes are found, including within the abdomen. It may indicate a primary lymphoma or be an indication of systemic inflammatory responses. Evaluation of the inciting cause will help in determining treatment options or indications for euthanasia.
Reactions to injections : Seen with use of some adjuvants, result in small subcutaneous lumps at the local site, which may ulcerate to a small dry, open lesion in the skin. In most cases no treatment is necessary; however guidelines should be established in studies for treatment if these lesions become larger or deeper than expected. They are usually sterile, so the primary concern is to prevent secondary infection.

Eyes and surrounding tissues

Problems of the eyes and surrounding tissues are commonly seen in clinical evaluation of mice. These may involve the structures in front of the eye, eyelids, conjunctiva, tear production, or tear drainage; the eye itself including the cornea, lens, and deep structures; or the orbit behind the eye. Blepharospasm (squinting), discharge from the eye, or buphthalmia (bulging) are the most common initial presentations.

Microphthalmia or anophthalmia : Congenital conditions (common in C57BL/6) which present as a partially opened or closed eye. Often tear production continues with poor drainage resulting in a mild watery or waxy ocular discharge. Most mice are stable and groom to keep the area clean, precluding the need for treatment. Occasionally treatment for conjunctivitis may be needed.
Conjunctivitis : Presents as swollen pink to red tissue under the eyelid and often a thick ocular discharge. It may be caused by foreign bodies such as a piece of bedding or an aberrant eyelash, or be related to trauma to the conjunctiva or the globe. Often this can be treated by gentle flushing of the eye with a saline eye wash and topical application of an antibiotic lubricant ointment. Systemic antibiotics are also beneficial.
Keratitis : Inflammation of the cornea, presents as a cloudy or vascular surface of the eye, and is often combined with conjunctivitis. Lack of tear production or inability to close the eyelids properly can lead to drying of the corneal surface. Ulceration of the cornea may be secondary to drying, or a result of trauma or displacement of the lens of the eye. A dent in the cornea may be visible and a tissue plug may be present if all layers of the cornea have been penetrated. The cornea is well innervated, so corneal lesions have potential to be quite painful. Treatment with an analgesic either topically or systemically combined with lubrication and antibiotics leads to healing of most such lesions. In severe cases the globe may collapse or be lost; however healing of the orbit can occur and the mouse itself can be maintained.
Cataracts : Develop in some strains of mice, such as C57BL/6. They are seen as a central white material behind a clear cornea. Generally they do not cause any problem for the animal, however occasionally the lens will luxate leading to inflammation within the eye, glaucoma, or even expulsion of the lens. In such cases treatment is the same as for keratitis. White scars on the cornea may be confused with cataracts.
Retro-orbital tumors , blood clots, or abscesses: Result in bulging of the eye forward of the normal position, and often difficulty in closing the eyelids. These are difficult to treat, and euthanasia of the mouse is usually the preferred option.

Mobility issues

Mobility issues may be due to injuries, central nervous system disorders, or degenerative conditions.

Injuries :The most common include catching a foot or leg in some part of the cage apparatus and fight wounds. Each instance needs to be evaluated with the veterinary team for appropriate pain management, treatment, or euthanasia.
Neurologic conditions : Present in many ways including ataxia, head tilt, spinning when lifted by the tail, circling with inability to straighten out the path, and seizures. The primary clinical concern is ability to reach feed and water. Supportive care includes placing a soft diet on the floor of the cage, soft bedding, support of bodily functions such as urination, etc. Maintenance of such mice should be justified in the study proposal, or they should be euthanized.
Arthritis : Presents as swelling and often redness of joints, with favoring of the affected limb or a reluctance to move. This may be transient, related to the strain or study, or a result of trauma. Analgesia is usually indicated, and if pain is not resolved then euthanasia may be necessary.
Pododermatitis : Irritation on the bottoms of the feet may be part of a study model using foot pad injections, or may be caused by the floor of the cage (wire floors) or a wet cage. Treatment will depend on the specifics of the case but may include antibiotics, analgesics, and soft absorbent bedding.

Respiratory issues

Respiratory problems are seen as changes in the breathing pattern or nasal discharge. Signs of respiratory distress include dyspnea, shallow rapid breathing, gasping, or abdominal effort in breathing. Nasal discharge may be seen dried on the nostrils, or more commonly as crusty material on the forelegs from self grooming.

Immunosuppressed animals are highly susceptible to pulmonary infections similar to those seen in humans with AIDS. Use of sterile caging greatly reduces the risk for these animals. Preventive strategies are most effective; once symptoms develop there is little chance for successful treatment and euthanasia is the preferred option.

Congenital deformities

Congenital deformities are generally seen shortly after birth or at the time of weaning. These conditions may also lead to difficulty with pregnancy or pup delivery by the dam. If not part of the research model, it is best not to use affected mice or their parents for future breeding of a research line. These problems include:

Hydrocephalus : A condition in which fluid builds up in the ventricles of the brain and does not distribute normally between the brain and spinal cord. Visibly these mice have a large rounded head and shortened muzzle. They are smaller than littermates, and with time develop lethargy and neurologic abnormalities. Supportive care with special feed may be provided short term, but these animals rarely survive to adulthood. Euthanasia is the most humane option.
Malocclusion : Mouse teeth grow throughout life. The teeth should meet in such a way that they grind on each other and on the feed to keep the teeth a normal length. When this does not happen teeth may grow into the palate or out of the mouth making eating or drinking difficult for the animal. Deformities may be caused by congenitally defective jaw structure, damage to the developing teeth, or trauma to the mouth or jaw. Treatment short term involves trimming of the incisor teeth, however care for crooked or maloccluded teeth is a lifelong process. Euthanasia should be strongly considered and breeding is not recommended.
Runt pups : Very small, poorly developing pups usually indicate a genetic abnormality, or competitive disadvantage. In very large litters or if the dam is a poor milk producer, supportive care with soft dough or gel diets may provide sufficient support for runts to catch up to normal mice. However, if a dam produces runts in subsequent litters, it is best to retire her from breeding.
Imperforate vagina : A defect that may occur in the maturing young female mouse. It is produced by lack of opening of the vaginal membrane, and appears as swelling between the anus and genital papilla giving the appearance of a male mouse. Female mice have nipples while males do not, which helps in determining the sex of these mice. Although the vaginal canal can be surgically opened, breeding performance is usually poor, so this defect also is an indication for euthanasia.
Other common congenital deformities : Extra limbs or toes, lack of one or more limbs, an outwardly curved sternum (breast bone), small or absent eyes, abnormal organ development which may lead to difficulty breathing or a distended abdomen, closed rectum leading to inability to defecate, and others. Each should be evaluated by the veterinary staff with the investigator to determine the significance to the mouse, the line, and the best clinical approach.

Reproductive associated conditions

Reproductive-associated conditions are a common reason for veterinary care in mouse colonies. Clinical success requires rapid identification; thus observations by investigators as well as the animal care team can be very helpful in addressing these situations.

Dystocia : Difficulty in delivery of pups is one of the most common and clinically difficult conditions in mice. Signs of dystocia include a pup visible in the vaginal canal but not passing, immobility and dehydration, distension of the abdomen with little muscle tone, or labor for an extended period of time (more than a couple hours). Mice often deliver their pups during the night, so early morning health checks are the most common time to discover dystocia. Causes may include congenitally deformed or dead pups, very large pups, breach presentation, non-dilated vaginal canal, or exhaustion of the dam. A stuck pup may be removed by gently applying a lubricant around the pup, grasping the pup with gauze, and exerting gentle traction on the pup. However, if the mother is weak, she is unlikely to push out additional pups. Caesarian delivery of the pups and fostering to another breeder with pups the same age or slightly older is recommended for valuable lines. Provision of warm fluids subcutaneously, soft diets, and treatment with oxytocin and calcium may be of benefit.
Prolapse of vaginal or uterine tissue : May be secondary to vaginal hyperplasia, or excessive abdominal contractions. If minor, the exposed tissue may be cleaned, treated with a hyperosmotic solution to reduce swelling, and replaced via the vagina. A suture may be used to close the vaginal opening for a few days (mice have a separate external urethral opening so closure of the vagina temporarily is okay). However, if the amount of tissue is large or there is evidence of necrosis or self mutilation then euthanasia is indicated.
Prolapsed penis (paraphimosis) : Occurs in male mice when the penis is not retracted into the surrounding prepuce. The usual presentation is a swollen, dragging penis, often with secondary trauma to the surface skin. Blockage of the urethra may also be noted. If the mouse is able to urinate, lubrication and placement on a soft bedding surface may allow the swelling to decrease and the penis to return to the normal position.
Perineal cysts : The bulbo-urethral glands may become filled with fluid giving the appearance of a severely enlarged scrotum or perineum. Needle aspiration yields a clear slightly yellow fluid. Generally no treatment is needed.

General conditions

Other general conditions that may be noted include:

Diarrhea : Noted as liquid feces when the animal is picked up or seen in the cage bedding. Diarrhea can lead to dehydration, so treatment is similar. Antibiotic therapy may also be beneficial.
Ascites : The buildup of fluid in the abdomen may be induced by a study; in which case the inclusion of endpoint guidelines is critical. It can also indicate organ failure of the heart or liver, neoplasia, or lymphatic malfunction. There is no long term treatment for this condition.
Anasarca : The buildup of fluid in the subcutaneous space, also indicative of organ failure, usually renal or lymphatic, is another indication for euthanasia. In examining a mouse this may initially be confused with obesity. Pitting edema, lethargy, and distribution of subcutaneous fluid will aid in differentiation of these conditions.
Rectal prolapse : A bulging of the distal colon out of the rectum, common in mice affected by Helicobacter spp. or intestinal parasites; but also caused by straining, constipation, or unspecific reasons. The health status of the colony will help in determining possible causes. Some respond well to hyperosmotic soaks, but many remain chronic and are managed by cleansing the exposed tissue, providing soft bedding, and in some cases using antibiotic and lubricant treatments as well.
Anorexia : While not directly observable, this is indicated when there is a lack of feces in a cage that has not just been cleaned, when there is no evidence of mice chewing on the chow, or when mice appear too thin or dehydrated. The first thing to check is the animal’s teeth for possible malocclusion then palpate for any masses in the abdomen and generally examine the mouse. Supportive care includes fluid therapy and feed on the floor.

Background characteristics

Background strain/stock characteristics include clinical presentations such as deafness, blindness, and hyperactivity. When choosing a research model or establishing a new genetically engineered line, these need to be considered. See Table 2 .

Clinical Presentations Associated with Strain or Background

Strain or Stock	Predisposed to conditions
C57BL/6	Hydrocephalus, Microphthalmia, Anopthalmia, Age related hearing loss, Malocclusion, Barbering, Ulcerative dermatitis
BALB/c	Male aggression, Heart ventricular mineralization, Corneal opacities, Conjunctivitis, Blepharitis, Periorbital abscesses, Age related hearing loss
C3H/He	Blindness, Corneal opacities, Age related hearing loss, Mammary tumors
FVB/N	Blindness, Seizures, Mammary hyperplasia (tumors rare), Hyperactivity
129	Blepharitis, Conjunctivitis, Megaesophagus
Swiss	Retinal degeneration, Amyloidosis
SJL/J	Blindness
A/J	Early hearing loss
DBA/2J	Audiogenic seizures, Early hearing loss

Developed from information in: Hedrich, H.J., Bullock, G., and Petrusz, P., 2004 ; Percy, D.H., and Barthold, S.W., 2007

DEFINING AND REFINING ENDPOINTS

Important for refining the way we conduct animal research is identifying appropriate endpoints, and facilitating ways to adhere to those endpoints in practice. Endpoints are meant to minimize or eliminate pain or distress, when possible, and enhance animal well being. The Guide defines a humane endpoint as “(T)he point at which pain or distress in an experimental animal is prevented, terminated or relieved” ( National Research Council, 2011 ).

Frequent health evaluations may be required to identify animals approaching a study’s endpoint, and these observations play an important role in assuring humane animal research. In some instances, the endpoint described in animal study proposals is a compromise between the humane endpoint and the experimental endpoint, the time at which scientific aims and objectives are met ( National Research Council, 2011 ). The Principal Investigator, in collaboration with a veterinarian and the Institutional Animal Care and Use Committee (IACUC), is responsible for identifying a study endpoint that is both scientifically relevant and humane before animal studies commence( Morton and Griffiths, 1985 ).

Morbidity endpoints are preferred and considered more humane than moribundity or death endpoints because they allow interventions or treatments that prevent pain and distress. However, in cases when veterinary intervention interferes with experimental results, moribundity/death as an endpoint may be required to reach experimental goals. Some examples of scientifically justified moribundity/death models include: metastatic tumor models, infectious disease/vaccine challenge, pain modeling, trauma, production of monoclonal antibodies, toxicology, organ/system failure, and cardiovascular shock ( National Research Council, 2011 ).

Identifying Animals Nearing Endpoint

Multiple criteria may indicate an animal has reached its study endpoint. While it is possible to describe an exhaustive list, Table 3 presents commonly referenced categories and criteria that can be used when planning a study and deciding on an appropriate study endpoint. It is not uncommon for an animal to show multiple clinical signs listed in the protocol while remaining below a study’sdescribed endpoint. In such cases, evaluation of the five criteria of an animal's condition as described by Morton and Griffiths is useful. These are: body weight, physical appearance, measurable clinical signs, unprovoked behavior and response to external stimuli ( Morton and Griffiths,1985 ). These criteria, when considered with objective measurements and in consultation with a veterinarian, can help identify animals at the earliest endpoint. It is also important to note that clinical signs may arise spontaneously that are not described in an approved study protocol. Attending veterinarians may, based on the animal’s overall condition, consider an animal to be at its humane endpoint and recommend euthanasia.

Clinical Signs and Evaluation Criteria Used to Determine Humane Endpoints

Clinical Signs (NIH, ARAC, 2011)	Clinical Observations in Cancer Research and Toxicological Studies ( )

Special Considerations of Endpoints

Endpoint evaluation criteria presented in Table 3 is a good foundation for evaluating laboratory mice. Some conditions, such as aging studies warrant extra consideration when defining study endpoints. Aging mice often exhibit a host of clinical signs that would indicate disease in younger mice (Office of Animal Care and Use, National Institutes of Health, 2011), including decreased body condition, increased respiratory rate, and pallor; but these may be considered normal in an aging mouse. More subjective criteria, such as quality of life and general health may be used.

Genetically Modified Animals (GMA) is an example of studies where unexpected experimental outcomes may occur. Small pilot studies and additional screening/monitoring are recommended since the effects of the genetic modifications can be largely unknown ( Stokes, 2000 ).One should also consider the use of body condition scores, rather than body weight, in the determination of endpoints in models involving tumors, ascites, and other diseases that cause organomegally. These conditions all can falsely increase body weight, despite concurrent muscle wasting and cachexia.

Importance of Training and Monitoring

Adequate training of staff to recognize clinical signs and identify animals at endpoint is key to minimizing pain and distress. When the nature or severity of an animal’s condition is in question, consultation with a veterinarian is useful not only in determining a diagnosis but in training oneself and the staff to recognize various disease states. Additionally, there are numerous training resources available to help staff become familiar with these clinical signs, such as the Charles River Handbook of Clinical Signs in Rodents and Rabbits ( Pritchett-Corning, 2010 ) .

The use of study specific records that detail both the assessment criteria for endpoints as well as the animal’s condition can play a key role in the effective monitoring of animals. Such a document would ideally include: the definition of the study endpoint and assessment criteria, frequency of observation, and the response required when the animal has reached the study endpoint (Office of Animal Care and Use, National Institutes of Health, 2011). Animal users and animal care staff can communicate more efficiently, and animals can be identified for intervention/euthanasia at the earliest possible time point, preventing or alleviating unnecessary pain or distress--a refinement by definition.

In summary, when we choose endpoints thoughtfully and use trained staff to monitor animals frequently, we improve the welfare of laboratory mice by minimizing pain or distress. Using training resources, key references on endpoints, and study-specific monitoring records lend to our mission of refining the way we do research on animals.

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Fight wounds. (A) Typical pattern of miliary wounds on the side of the body. (B) Deep wound on the caudal portion of the rump. (C) Wounds and associated swelling on the tail.

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Ear Dermatitis. Crusty lesions on and below the ear associated with loss of an eartag.

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Barbering. (A) Thin coat with short stubby hairs on the head and neck. (B) Two mice, one with minor and one with extensively barbered fur. Note that the skin is healthy in these cases.

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Ulcerative dermatitis, deep ulcerative lesion with redness and moist surface at the base of the neck.

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Ringtail. Circular constriction is noted with normal skin coloration. There may be one ring as shown or a series of rings around the tail.

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Tumor. A subcutaneous irregular mass is shown caudal to the front leg of this nude mouse.

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Conjunctivitis. The left eye demonstrates swelling of the eyelids, redness of conjunctiva, and serous discharge. Compare to the normal right eye.

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Keratitis. The left cornea is white, opaque, and dry. Compare to the normal right eye.

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Retrobulbar abscess. Swelling and discoloration are noted below and caudal to the orbit of the left eye. Compare to the normal right eye.

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Arthritis. The mouse on the left has swelling of the hock (ankle) joint associated with arthritis. A normal mouse is shown on the right.

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Malocclusion. The incisor teeth are unequal in length and the shorter tooth angles inward more than normal. Teeth may be observed to grow inward or outward, and may be very curved or broken as shown.

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Runts. A runt or small mouse is shown on the left compared with its normal littermate on the right.

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Imperforate vagina. Both are female mice. The mouse on the left lacks the normal vaginal opening and shows subcutaneous perineal swelling due to accumulated secretions which are unable to drain. Normal mouse is shown on the right.

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Bulbourethral gland cyst. Swelling in the right side of the scrotum is caused by a cystic bulbourethral gland. This can be confirmed by performing a needle aspiration, with fluid indicating a cyst while no fluid may be indicative of a tumor.

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Ascites. Fluid accumulation within the abdomen leads to a potbellied appearance with prominent spine.

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Rectal prolapse. Red, edematous mucosal tissue is seen bulging from the rectal orifice.

Internet Resources with Annotations:

http://phenome.jax.org Mouse phenome database maintained by The Jackson Laboratory with detailed phenotype strain survey data. The Jackson Laboratory site www.jax.org has links to many valuable resources such as mouse nomenclature, resource manuals, and specific strain information.

http://www.ors.od.nih.gov/sr/dvr/od/Documents/Post_Op_Form.docx A link to a post-op monitoring form that may be useful to either the investigative team and/or the veterinary team.

An example of commercial laboratory services for health surveillance can be found at: http://www.criver.com/en-US/ProdServ/ByType/ResAnimalDiag/Pages/home2.aspx

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Title: “Health Evaluation of Experimental Laboratory Mice”

Legend: Instructional video detailing normal behavior, home cage observation, restraint, body conditioning, and physical examination of experimental laboratory mice.

File name: Circling in Laboratory Mice.mov

Title: “Circling in Laboratory Mice”

Legend: An example of circling in a laboratory mouse.

File name: Ataxia in Laboratory Mice.mov

Title: “Ataxia in Laboratory Mice”

Legend: An example of ataxia in a laboratory mouse.

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Williams-Fritze MJ, Carlson-Scholtz JA, Zeiss C, Deng Y, Wilson SR, Franklin R, Smith PC. Maropitant citrate for treatment of ulcerative dermatitis in mice with a C57BL/6 background. J. Am. Assoc. Lab. Anim. Sci. 2011; 50 (2):221–226. [ PMC free article ] [ PubMed ] [ Google Scholar ]

Key References with Annotations

Hedrich HJ, Bullock G, Petrusz P. The Laboratory Mouse. San Diego, CA: Elsevier Academic Press; 2004. [ Google Scholar ] . This book contains excellent introductory information in first few chapters with strain characteristics in Chapter 3. Detailed systems information excellent as a reference.
Percy DH, Barthold SW. Pathology of Laboratory Rodents and Rabbits. Third edition. Ames, IA: Blackwell Publishing; 2007. [ Google Scholar ] . The first chapter of this book contains excellent general pathology information on laboratory mice, including strain characteristics.
Pritchett-Corning KR, Girod A, Avellaneda G, Fritz PE, Chou S, Brown MJ. Handbook of Clinical Signs in Rodents and Rabbits. Charles River Laboratories; 2010. www.criver.com [ Google Scholar ] . This is a pictorial guidebook aimed at animal care personnel, organized by anatomic and organ system sections which has additional information on rodent clinical observations.

Call for Nominations: Harvard Medical School 2025 Warren Alpert Foundation Prize

URL: https://warrenalpert.org/

DESCRIPTION:

The Warren Alpert Foundation Prize, in association with Harvard Medical School, recognizes and honors one or more scientists, physicians and researchers whose scientific achievements have led to the prevention, cure, or treatment of human disorders or for seminal research that holds great promise to change our ability to treat disease.

The prize was established in 1987 by the late philanthropist and businessman, Warren Alpert and the Warren Alpert Foundation. The Warren Alpert Prize is given internationally and since its inception, ten Nobel Prize winners have received the award. The prize is administered in concert with Harvard Medical School in Boston, Massachusetts and the Warren Alpert Foundation, located in Providence, Rhode Island. An annual scientific symposium is held at Harvard Medical School each fall in honor of the recipient(s).

ELIGIBILITY:

Up to five nominees may be co-nominated in one nomination. Multiple nominations from an institution are permitted.

Nominations are accepted from physicians and scientists from around the world. U.S. and non-U.S. citizens are eligible to be nominated.

NOMINATION PROCESS:

Awards include $500,000 (to be split equally if more than one recipient is selected), a citation and plaque. Nominations are submitted through an online form. Nominations must include:

Contact information for the nominee and nominator.
Optional additional nominees (if applicable).
A suggested citation for the award (maximum fifty words).
Description of the nominee’s research as it relates to the nomination. Include the top five bibliographical references related to the nominations.
If nominating an individual, comment on whether it is appropriate to share this prize. If so, please describe their contributions.
Two letters of support.
CV of the nominee(s).

Nominations Due by Friday, November 8, 2024

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Methamphetamine-involved psychiatric hospitalizations have increased, study says

by CU Anschutz Medical Campus

A new study, out now in Drug and Alcohol Dependence , that details trends among psychiatric hospitalizations between 2015-2019 finds that while most hospitalizations did not involve any substances, methamphetamine-related hospitalizations have increased while overall number of psychiatric hospitalizations remained stable.

Additionally, researchers detail that psychiatric hospitalizations caused by methamphetamine use were highest in the Mountain West region but were also shifting geographically. "Rates of methamphetamine-involved psychiatric hospitalizations with were by far the highest in the Mountain West. As expected, this mirrors rates of self-reported methamphetamine use and methamphetamine-related overdose deaths in the Mountain West." says Susan Calcaterra, MD, MPH, professor at the University of Colorado Anschutz Medical Campus and study lead author. "Psychiatric hospitalizations involving methamphetamine use is really taking off in the Midwest and Northeast, in particular."

While rates of methamphetamine-related psychiatric hospitalizations increased 68% over the study period, opioid-related hospitalizations decreased by 22%. Methamphetamine rate increases may be attributed to methamphetamines ubiquitousness and affordability, as well as the lack of resources available to manage methamphetamine use. Why opioid-involved psychiatric hospitalizations declined is less clear but may be related to the lethality of fentanyl.

"An important takeaway from this study is the need for resources to address the mental and physical treatment of methamphetamine use," says Calcaterra.

"While the vast majority of psychiatric hospitalizations in this timeframe did not involve substance use, the significant increase in methamphetamine use means we have to better consider harm reduction in clinical settings. Evidence-based interventions such as contingency management which involves offering incentives for abstinence, harm reduction education, provision of naloxone for overdose reversal and access to expanded mental health treatments are proven to help mitigate dangerous effects from methamphetamine use, especially when contaminated with fentanyl much like the campaigns aimed at public awareness around opioid use."

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Biden announces $150 million in research grants as part of his ‘moonshot’ push to fight cancer

President Joe Biden promotes his “moonshot” initiative aimed at reducing cancer deaths in New Orleans. The president announces $150 million in awards from the Advanced Research Projects Agency for Health supporting eight research teams around the country.

President Joe Biden and first lady Jill Biden listen during a demonstration of cancer research and detection techniques at Tulane University, Tuesday, Aug. 13, 2024, in New Orleans. (AP Photo/Mark Schiefelbein)

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President Joe Biden listens as Tulane University President Michael Fitts speaks during a demonstration of cancer research and detection techniques at Tulane University, Tuesday, Aug. 13, 2024, in New Orleans. (AP Photo/Mark Schiefelbein)

President Joe Biden greets former New Orleans Mayor Mitch Landrieu and his wife Cheryl Tuesday, Aug. 13, 2024, at Louis Armstrong International Airport in New Orleans. (AP Photo/Mark Schiefelbein)

President Joe Biden talks with reporters Tuesday, Aug. 13, 2024, at Louis Armstrong International Airport in New Orleans. (AP Photo/Mark Schiefelbein)

President Joe Biden speaks to reporters as he departs the White House for a trip to New Orleans, Tuesday, Aug. 13, 2024, in Washington. (AP Photo/Manuel Balce Ceneta)

FILE - President Joe Biden speaks on the cancer moonshot initiative at the John F. Kennedy Library and Museum, Sept. 12, 2022, in Boston. (AP Photo/Evan Vucci)

President Joe Biden speaks to reporters as he leaves the White House for a trip to New Orleans, La., Tuesday, Aug. 13, 2024, in Washington. (AP Photo/Manuel Balce Ceneta)

President Joe Biden and first lady Jill Biden board Air Force One as they arrive to depart, Tuesday, Aug. 13, 2024, at Joint Base Andrews, Md., en route to New Orleans. (AP Photo/Mark Schiefelbein)

President Joe Biden, escorted by Air Force Col. Angela Ochoa, Commander, 89th Airlift Wing, walks to Air Force One as he arrives to depart, Tuesday, Aug. 13, 2024, at Joint Base Andrews, Md., en route to New Orleans. (AP Photo/Mark Schiefelbein)

NEW ORLEANS (AP) — President Joe Biden is zeroing in on the policy goals closest to his heart now that he’s no longer seeking a second term , visiting New Orleans on Tuesday to promote his administration’s “moonshot” initiative aiming to dramatically reduce cancer deaths.

The president and first lady Jill Biden toured medical facilities that receive federal funding to investigate cancer treatments at Tulane University. Researchers used a piece of raw meat to demonstrate how they are working to improve scanning technology to quickly distinguish between healthy and cancerous cells during surgeries.

The Bidens then championed the announcement of $150 million in awards from the Advanced Research Projects Agency for Health. Those will support eight teams of researchers around the country working on ways to help surgeons more successfully remove tumors from people with cancer. It brings the total amount awarded by the agency to develop breakthrough treatments for cancers to $400 million.

Cancer surgery “takes the best surgeons and takes its toll on families,” Biden said. He said the demonstration of cutting-edge technology he witnessed would offer doctors a way to visualize tumors in real time, reducing the need for follow-on surgeries.

“We’re moving quickly because we know that all families touched by cancer are in a race against time,” Biden said.

The teams receiving awards include ones from Tulane, Dartmouth College, Johns Hopkins University, Rice University, the University of California, San Francisco, the University of Illinois Urbana-Champaign, the University of Washington and Cision Vision in Mountain View, California.

Before he leaves office in January, Biden hopes to move the U.S. closer to the goal he set in 2022 to cut U.S. cancer fatalities by 50% over the next 25 years, and to improve the lives of caregivers and those suffering from cancer.

“I’m a congenital optimist about what Americans can do,” Biden said. “There’s so much that we’re doing. It matters”

Experts say the objective is attainable — with adequate investments.

“We’re curing people of diseases that we previously thought were absolutely intractable and not survivable,” said Karen Knudsen, CEO of the American Cancer Society and the American Cancer Society Cancer Action Network.

Cancer is the second-highest killer of people in the U.S. after heart disease. This year alone, the American Cancer Society estimates that 2 million new cases will be diagnosed and 611,720 people will die of cancer diseases.

Still, “if all innovation ended today and we could just get people access to the innovations that we know about right now, we think we could reduce cancer mortality by another 20 to 30%,” Knudsen said.

The issue is personal enough for Biden that, in his recent Oval Office address about bowing out of the 2024 campaign, the president promised to keep fighting for “my cancer moonshot so we can end cancer as we know it.”

“Because we can do it,” Biden said then.

He said in that speech that the initiative would be a priority of his final months in office, along with working to strengthen the economy and defend abortion rights, protecting children from gun violence and making changes to the Supreme Court, which he called “extreme” in its current makeup during a recent event.

Both the president and first lady have had lesions removed from their skin in the past that were determined to be basal cell carcinoma, a common and easily treated form of cancer. In 2015, their eldest son, Beau, died of an aggressive brain cancer at age 46.

“It’s not just personal,” Biden said Tuesday. “It’s about what’s possible.”

The president’s public schedule has been much quieter since he left the race and endorsed Vice President Kamala Harris , making Tuesday’s trip stand out.

Advocates have praised Biden for keeping the spotlight on cancer, bringing stakeholders together and gathering commitments from private companies, nonprofit organizations and patient groups.

They say that the extra attention the administration has paid has put the nation on track to cut cancer death rates by at least half, preventing more than 4 million deaths from the disease, by 2047. It has done so by bolstering access to cancer treatments and reminding people of the importance of screening, which hit a setback during the coronavirus pandemic.

“President Biden’s passion and commitment to this effort has made monumental differences for the entire cancer community, including those who are suffering from cancer,” said Jon Retzlaff, the chief policy officer at the American Association for Cancer Research.

Looking ahead, Retzlaff said, “The No. 1 thing is for us to see robust, sustained and predictable annual funding support for the National Institutes of Health. And, if we see that through NIH and through the National Cancer Institute, the programs that have been created through the cancer moonshot will be allowed to continue.”

Initiatives under Biden include changes that make screening and cancer care more accessible to more people, said Knudsen, with the American Cancer Society.

For instance, Medicare has started to pay for follow-up colonoscopies if a stool-based test suggests cancer, she said, and Medicare will now pay for navigation services to guide patients through the maze of their cancer care.

“You’ve already paid for the cancer research. You’ve already paid for the innovation. Now let’s get it to people,” Knudsen said.

She also said she’d like to see the next administration pursue a ban on menthol-flavored cigarettes, which she said could save 654,000 lives over the next 40 years.

Scientists now understand that cancer is not a single disease, but hundreds of diseases that respond differently to different treatments. Some cancers have biomarkers that can be targeted by existing drugs that will slow a tumor’s growth. Many more targets await discovery.

“We hope that the next administration, whoever it may be, will continue to keep the focus and emphasis on our national commitment to end cancer as we know it,” said Dr. Crystal Denlinger, CEO of the National Comprehensive Cancer Network, a group of elite cancer centers.

Johnson reported from Washington state.

IMAGES

Laboratory rats, mice and rodent video capture for biomedical research
Why Mice are Ideal For Testing and Research
Medical Research Scientists Examines Laboratory Mice kept in a G
Mice in medical research
Biotechnology Laboratory. Mouse for animal experiment. DNA, cancer
Medical Research Scientists Examines Laboratory Mice kept in a Glass

COMMENTS

The Mighty Mouse: The Impact of Rodents on Advances in Biomedical Research
Mice and rats have long served as the preferred species for biomedical research animal models due to their anatomical, physiological, and genetic similarity to humans. Advantages of rodents include their small size, ease of maintenance, short life cycle, ...
Why Mice for Biomedical Research?
With mice, researchers can readily track the genetics that underlie those differences and use their findings to inform drug development, and more accurate clinical trials. Mice are the key filling in the blanks of human genomics, and their presence in research is vital for the development of new diagnostics, treatments, and preventative actions.
The Applicability of Mouse Models to the Study of Human Disease
The chapter concludes with a discussion on the future of using mice in medical research with regard to ethical and technological considerations. Key words: Mouse, Model, Disease, Genetics, Physiology, Immunology, Ethics
Why are mice excellent models for humans?
Why are mice considered excellent models for humans? Humans and mice don't look alike, but both species are mammals and are biologically very similar. Almost all of the genes in mice share functions with the genes in humans. That means we develop in the same way from egg and sperm, and have the same kinds of organs (heart, brain, lungs, kidneys ...
Our mice, our hope
Our mice, our hope The use of mice in biomedical research has contributed dramatically to medical progress. In fact, medicine today is built on a foundation of mice as models of human disease. Mice are biologically similar to us, get most of the same diseases with the same genetic susceptibilities, and can be genetically manipulated to mimic most human diseases and conditions.
Mice make medical history
An army of mice, perhaps 25 million strong, each day help researchers worldwide to study and devise treatments for human ailments such as cancer, heart disease, AIDS and malaria. Mice are helping ...
NIH to make a mightier mouse resource for understanding disease
NIH to make a mightier mouse resource for understanding disease Over the next five years, National Institutes of Health (NIH)-funded researchers will extensively test and generate data about mice with disrupted genes to gain clues about human diseases. NIH today awarded a set of cooperative agreements totaling more than $110 million to begin the second phase of the Knockout Mouse Project (KOMP).
Advances in Transgenic Mouse Models to Study Infections by Human
This review discusses progress in the development and use of transgenic mice for the study of virus-induced human diseases towards identification of new drug innovations to treat and control human pathogenic infectious diseases. Keywords: human pathogenic viruses, transgenic mice, humanized mouse models. 1.
The Applicability of Mouse Models to the Study of Human Disease
The chapter concludes with a discussion on the future of using mice in medical research with regard to ethical and technological considerations. Keywords: Disease; Ethics; Genetics; Immunology; Model; Mouse; Physiology.
The Case for Female Mice in Neuroscience Research
Findings reveal female mice have more stable exploratory behavior than male mice. Mice have long been a central part of neuroscience research, providing a flexible model that scientists can control and study to learn more about the intricate inner workings of the brain. Historically, researchers have favored male mice over female mice in ...
Mice in medical research
Mice are versatile, they are used in a range of research from genetics to virology, oncology and many more. Notably, mice and other animals have been very important in the development of Herceptin, a monoclonal antibody used in certain types of breast cancer. Herceptin was the first monoclonal antibody successfully used to treat cancer.
Why we need female mice in neuroscience research
Findings reveal that despite hormonal fluctuations, female mice exhibit more stable exploratory behavior than their male peers Mice have long been a central part of neuroscience research, providing a flexible model that scientists can control and study to learn more about the intricate inner workings of the brain. Historically, researchers have favored male mice over female mice in experiments ...
Major New Study Reveals New Similarities and Differences Between Mice
A new, comprehensive study of the mouse genome by an international group of researchers including Penn State University scientists reveals striking similarities and differences with the human genome. The study may lead to better use of mouse models in medical research.
Why Do Medical Researchers Use Mice?
In fact, 95 percent of all lab animals are mice and rats, according to the Foundation for Biomedical Research (FBR). Scientists and researchers rely on mice and rats for several reasons. One is ...
Mice in scientific research
In the UK, mice are the most used animal in biomedical research. In this video we explain why and how mice are used in research, and how they're looked after...
Rewinding the Clock
The answer appears to be yes, at least in mice, according to a new study led by investigators at Harvard Medical School. The research, published March 22 in Cell, identifies the key cellular mechanisms behind vascular aging and its effects on muscle health and has successfully reversed the process in animals.
What is a mouse model?
JAX pioneered the use of mice in disease research, and its mice and research program have contributed to important medical breakthroughs ever since. Throughout, JAX has spearheaded the drive for increasing the accuracy and relevance of mouse-based biomedical research.
Medical Research Using Animals Often Fails To Produce Drugs That Work
Most potential new drugs don't work when tested in people. It's a major disappointment and it drives up the cost of developing new drugs. One big reason is the use of animals in medical research.
Molecule restores cognition and memory in Alzheimer's disease mouse
To test whether this would result in improved memory and cognition, researchers used mice that were genetically modified to have symptoms of Alzheimer's disease. Both these Alzheimer's disease-model mice and wild-type mice underwent baseline cognitive testing in a Barnes maze — a circular platform surrounded by visual clues and containing ...
Molecule restores cognition and memory in Alzheimer's disease mouse
In a new study, a molecule identified and synthesized by UCLA Health researchers was shown to restore cognifitive functions in mice with Alzheimer's disease symptoms. The compound effectively jump-started the brain's memory circuity in the mice. If proven to have similar effects in humans, the ...
Molecule restores cognition, memory in Alzheimer's disease model mice
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The Applicability of Mouse Models to the Study of Human Disease

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Health Evaluation of Experimental Laboratory Mice

INTRODUCTION

COLONY HEALTH SURVEILLANCE

EXAMINATION AND ASSESSMENT OF THE MOUSE

Home cage evaluation

Handling and Hands on evaluation

Restraint and physical examination

COMMON CLINICAL HEALTH CONDITIONS OF MICE

Skin lesions

Lumps and Bumps

Eyes and surrounding tissues

Mobility issues

Respiratory issues

Congenital deformities

Reproductive associated conditions

General conditions

Background characteristics

DEFINING AND REFINING ENDPOINTS

Identifying Animals Nearing Endpoint

Special Considerations of Endpoints

Importance of Training and Monitoring

LITERATURE CITED

Key References with Annotations

Call for Nominations: Harvard Medical School 2025 Warren Alpert Foundation Prize

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