Laboratory mouse

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Laboratory mice
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albino
laboratory mouse is an iconic model organism for scientific research in a variety of fields
Albino SCID
An SCID
With intermediate coat colour
Intermediate coat colour
Kept as a pet standing on a patch of grass
Kept as a pet

The laboratory mouse or lab mouse is a small

scientific disciplines. Mice belong to the Euarchontoglires clade, which includes humans. This close relationship, the associated high homology with humans, their ease of maintenance and handling, and their high reproduction rate, make mice particularly suitable models for human-oriented research. The laboratory mouse genome has been sequenced and many mouse genes have human homologues.[1] Lab mice are sold at pet stores for snake food and can also be kept as pets
.

Other mouse species sometimes used in laboratory research include two American species, the

Peromyscus maniculatus
).

History as a biological model

Mice have been used in biomedical research since the 17th century when William Harvey used them for his studies on reproduction and blood circulation and Robert Hooke used them to investigate the biological consequences of an increase in air pressure.[2] During the 18th century Joseph Priestley and Antoine Lavoisier both used mice to study respiration. In the 19th century Gregor Mendel carried out his early investigations of inheritance on mouse coat color but was asked by his superior to stop breeding in his cell "smelly creatures that, in addition, copulated and had sex".[2] He then switched his investigations to peas but, as his observations were published in a somewhat obscure botanical journal, they were virtually ignored for over 35 years until they were rediscovered in the early 20th century. In 1902 Lucien Cuénot published the results of his experiments using mice which showed that Mendel's laws of inheritance were also valid for animals — results that were soon confirmed and extended to other species.[2]

In the early part of the 20th century,

fancy mice and rats which she marketed to rodent hobbyists and keepers of exotic pets, and later began selling in large numbers to scientific researchers.[3] Together they generated the DBA (Dilute, Brown and non-Agouti) inbred mouse strain and initiated the systematic generation of inbred strains.[4] The mouse has since been used extensively as a model organism and is associated with many important biological discoveries of the 20th and 21st Centuries.[2]

The Jackson Laboratory in Bar Harbor, Maine is currently one of the world's largest suppliers of laboratory mice, at around 3 million mice a year.[5] The laboratory is also the world's source for more than 8,000 strains of genetically defined mice and is home of the Mouse Genome Informatics database.[6]

Reproduction

1-day-old pups

Breeding onset occurs at about 50 days of age in both females and males, although females may have their first

copulatory plug in the vagina up to 24 hours post-copulation. The presence of sperm on a vaginal smear is also a reliable indicator of mating.[7]

The average gestation period is 20 days. A fertile

outbred and hybrid mice. The young are called pups and weigh 0.5–1.5 g (0.018–0.053 oz) at birth, are hairless, and have closed eyelids and ears. Pups are weaned at 3 weeks of age when they weigh about 10–12 g (0.35–0.42 oz). If the female does not mate during the postpartum estrus, she resumes cycling 2–5 days post-weaning.[7]

Newborn males are distinguished from newborn females by noting the greater

littermates and comparing perinea.[7]

Genetics and strains

Mice are mammals of the

flying lemurs
.

Euarchontoglires
Glires

Rodentia (rodents)

Lagomorpha (rabbits, hares, pikas)

Euarchonta

Scandentia (treeshrews)

Primatomorpha

Dermoptera (flying lemurs)

Haplorrhini
)

Laboratory mice are the same species as the

transgenic strains. A strain, in reference to rodents, is a group in which all members are as nearly as possible genetically identical. In laboratory mice, this is accomplished through inbreeding. By having this type of population, it is possible to conduct experiments on the roles of genes, or conduct experiments that exclude genetic variation as a factor. In contrast, outbred populations are used when identical genotypes are unnecessary or a population with genetic variation is required, and are usually referred to as stocks rather than strains.[8][9] Over 400 standardized, inbred strains have been developed.[citation needed
]

Most laboratory mice are hybrids of different subspecies, most commonly of Mus musculus domesticus and Mus musculus musculus. Laboratory mice can have a variety of coat colours, including agouti, black and

albino. Many (but not all) laboratory strains are inbred. The different strains are identified with specific letter-digit combinations; for example C57BL/6 and BALB/c. The first such inbred strains were produced in 1909 by Clarence Cook Little, who was influential in promoting the mouse as a laboratory organism.[10] In 2011, an estimated 83% of laboratory rodents supplied in the U.S. were C57BL/6 laboratory mice.[11]

Genome

Sequencing of the laboratory mouse

haploid genome is about three billion base pairs long (3,000 Mb distributed over 19 autosomal chromosomes plus 1 respectively 2 sex chromosomes), therefore equal to the size of the human genome.[citation needed] Estimating the number of genes contained in the mouse genome is difficult, in part because the definition of a gene is still being debated and extended. The current count of primary coding genes in the laboratory mouse is 23,139.[12] compared to an estimated 20,774 in humans.[12]

Mutant and transgenic strains

Two mice expressing enhanced green fluorescent protein under UV-illumination flanking one plain mouse from the non-transgenic parental line
Comparison of a knockout obese mouse (left) and a normal laboratory mouse (right)

Various mutant strains of mice have been created by a number of methods. A small selection from the many available strains includes -

  • Mice resulting from ordinary breeding and inbreeding:
    • diabetes mellitus type 1
      .
    • Murphy Roths large (MRL) mice, with unusual regenerative capacities[13]
    • Japanese waltzing mice, which walk in a circular pattern due to a mutation adversely affecting their inner ears
    • transplantation
      .
    • Severe combined immunodeficiency (SCID) mice, with an almost completely defective immune system
    • FVB mice
      , whose large litter sizes and large oocyte pronuclei expedite use in genetic research
    • recessive mutation tx which arose in an inbred. Theophilos et al. 1996 found this to be genetic and localized to chromosome 8, near the centromere.[14]
  • Transgenic mice, with foreign genes inserted into their genome:
  • Knockout mice, where a specific gene was made inoperable by a technique known as gene knockout: the purpose is to study the function of the gene's product or to simulate a human disease
    • Fat mice, prone to obesity due to a carboxypeptidase E deficiency
    • Strong muscular mice, with a disabled myostatin gene, nicknamed "mighty mice".

Since 1998, it has been possible to clone mice from cells derived from adult animals.

Commonly used inbred strains

There are many

mice used in research, however, inbred strains are usually the animals of choice for most fields. Inbred mice are defined as being the product of at least 20 generations of brother X sister mating, with all individuals being derived from a single breeding pair.[15]

Inbred mice have several traits that make them ideal for research purposes. They are

recessive traits that could cause problems.[16] They also have very unified phenotypes due to this stability.[16]

Many inbred strains have well documented traits that make them ideal for specific types of research. The following table shows the top 10 most popular strains according to Jackson Laboratories.

Common inbred strains of laboratory mice available from Jackson Laboratories
Strain Coat color[17] Common research uses Total Pubmed publications referencing the strain as of April 19, 2023[18]
C3HeB/FeJ Agouti Immunology, inflammation, autoimmunity[19] 482
NOD/ShiLtJ Albino Autoimmune type 1 diabetes[20] 105
DBA/1J Dilute brown Rheumatoid arthritis[21] 445
BALB/cByJ Albino Cancer, cardiovascular, immunology[22] 628
DBA/2J Dilute brown Cardiovascular, dermatology, developmental biology[23] 2,722
C3H/HeJ Agouti Cancer, cardiovascular, hematology[24] 4,037
C57BL/6J Black General purpose, background[25] 25,723
SJL/J Albino Cancer, cardiovascular, dermatology[26] 1,448
FVB/NJ Albino Immunology, inflammation, autoimmunity[27] 350
129S1/SvImJ Agouti Targeted mutations, cancer[28] 222

Jackson Labs DO project

Phylogenetic tree of the eight founder strains used in the DO project, as well as their approximate age of divergence. M. spretus is included as an outgroup that diverged ~2 million years ago (mya), it is not part of the DO project.[29]

The Jackson Labs DO (Diversity Outbred) project[30] is a mouse breeding program using multiple inbred founder strains to create a genetically diverse population of mice for use in scientific research.

These mice are designed for fine

genetic mapping, and capture a large portion of the genetic diversity of the mouse genome.[31]

This project has resulted in over 1,000 genetically diverse mice which have been used to identify genetic factors for diseases such as obesity, cancer, diabetes, and alcohol use disorder. [32]

Founder strains used in the DO project
Strain Derivation Subspecies origin Coat color[17] Common research uses Total Pubmed publications referencing the strain as of April 19, 2023
A/J Laboratory Mus musculus domesticus[33] Albino Cancer, immunology[34] 5,500
C57BL/6J Laboratory Mus musculus domesticus[33] Black General purpose, background[25] 25,723
129S1/SvImJ Laboratory Mus musculus domesticus Agouti[28] Targeted mutations, cancer[28] 222
NOD/ShiLtJ Laboratory Mus musculus domesticus[33] Albino Autoimmune type 1 diabetes[20] 105
NZO/HILtJ Laboratory Mus musculus domesticus[33] Agouti Obesity[35] 11
CAST/EiJ Wild-derived Mus musculus castanus[33] Agouti
genetic mapping[36]
 154
PWK/PhJ Wild-derived Mus musculus musculus [33] Agouti
Genetic mapping[37]
 52
WSB/EiJ Wild-derived Mus musculus domesticus[33] Agouti with head blaze, greyish coat
Genetic mapping, evolution[38]
 65

Appearance and behaviour

Laboratory mice have retained many of the physical and behavioural characteristics of house mice; however, due to many generations of artificial selection, some of these characteristics now vary markedly. Due to the large number of strains of laboratory mice, it is impractical to comprehensively describe the appearance and behaviour of all of them; however, they are described below for two of the most commonly used strains.

C57BL/6

A female C57BL/6 laboratory mouse
Main article: C57BL/6

C57BL/6 mice have a dark brown, nearly black coat. They are more sensitive to noise and odours and are more likely to bite than the more docile laboratory strains such as BALB/c.[39]

Group-housed C57BL/6 mice (and other strains) display barbering behaviour, in which the dominant mouse in a cage selectively removes hair from its subordinate cage mates.[40] Mice that have been barbered extensively can have large bald patches on their bodies, commonly around the head, snout, and shoulders, although barbering may appear anywhere on the body. Both hair and vibrissae may be removed. Barbering is more frequently seen in female mice; male mice are more likely to display dominance through fighting.[41]

C57BL/6 has several unusual characteristics which make it useful for some research studies but inappropriate for others: It is unusually sensitive to pain and to cold, and

morphine addiction, atherosclerosis, and age-related hearing loss.[11] When compared directly to BALB/c mice, C57BL/6 mice also express both a robust response to social rewards[43][44] and empathy.[45]

BALB/c

Main article: BALB/c
BALB/c laboratory mice

BALB/c is an

animal experimentation.[46]

BALB/c are noted for displaying high levels of anxiety and for being relatively resistant to diet-induced atherosclerosis, making them a useful model for cardiovascular research.[47][48]

Male BALB/c mice are aggressive and will fight other males if housed together. However, the BALB/Lac substrain is much more docile.[49] Most BALB/c mice substrains have a long reproductive life-span.[46]

There are noted differences between different BALB/c substrains, though these are thought to be due to

hermaphroditism.[51]

Tg2576

A useful model for

amyloid fibrils with aging, plaque formation, and impaired hippocampus learning and memory. Tg2576 mice are a good model for early-stage AD because they show amyloidogenesis and working memory impairments linked to age but do not show neuronal degeneration.[55] The absence of cell death suggests that changes in typical cellular signaling cascades involved in learning and synaptic plasticity are probably linked to the memory phenotype. Associative learning impairments are exacerbated when Tg2576 mice are crossed with PS1 transgenic animals that possess the A246E FAD mutation. This crosses promotes the build-up of amyloid and plaque development in the CNS.[56] This lends credence to the theory that AD pathogenesis is influenced by the interplay between APP and PS-1 gene products. Although Tg2576 mice do not perfectly replicate late-stage AD with cell death, they do offer a platform for researching the physiology and biochemistry of the illness.With the help of transgenic mouse models, researchers can make progress in AD research by understanding the intricate relationships between gene products that are involved in the production of Aβ peptide.e physiology and biochemistry of the illness.[57][58]

Husbandry

Laboratory mouse (note the ear tag)

Handling

Traditionally, laboratory mice have been picked up by the base of the tail. However, recent research has shown that this type of handling increases anxiety and aversive behaviour.[59] Instead, handling mice using a tunnel or cupped hands is advocated. In behavioural tests, tail-handled mice show less willingness to explore and to investigate test stimuli, as opposed to tunnel-handled mice which readily explore and show robust responses to test stimuli.[60]

Nutrition

In nature, mice are usually herbivores, consuming a wide range of fruit or grain.[61] However, in laboratory studies it is usually necessary to avoid biological variation and to achieve this, laboratory mice are almost always fed only commercial pelleted mouse feed. Food intake is approximately 15 g (0.53 oz) per 100 g (3.5 oz) of body weight per day; water intake is approximately 15 ml (0.53 imp fl oz; 0.51 US fl oz) per 100 g of body weight per day.[7]

Injection procedures

needle gauge
and recommended maximum injected volume at a single time at one site, as given in the table below:

Route
 Recommended site[62]
Needle gauge[62]
 Maximal volume[63]
subcutaneous dorsum, between scapula 25-26
ga
 2-3
ml
intraperitoneal
left lower quadrant
 25-27 ga 2-3 ml
intravenous
 lateral tail vein 27-28 ga 0.2 ml
intramuscular
 hindlimb, caudal thigh 26-27 ga 0.05 ml
intracerebral cranium 27 ga

To facilitate intravenous injection into the tail, laboratory mice can be carefully warmed under heat lamps to

vasodilate the vessels.[62]

Anaesthesia

A common regimen for

general anesthesia for the house mouse is ketamine (in the dose of 100 mg per kg body weight) plus xylazine (in the dose of 5–10 mg per kg), injected by the intraperitoneal route.[64] It has a duration of effect of about 30 minutes.[64]

Euthanasia

Approved procedures for

anesthetics, inhalable anesthetics, such as Halothane, and physical methods, such as cervical dislocation and decapitation.[65] In 2013, the American Veterinary Medical Association issued new guidelines for CO2 induction, stating that a flow rate of 10% to 30% volume/min is optimal for euthanasing laboratory mice.[66]

Pathogen susceptibility

A recent study detected a murine astrovirus in laboratory mice held at more than half of the US and Japanese institutes investigated.[67] Murine astrovirus was found in nine mice strains, including NSG, NOD-SCID, NSG-3GS, C57BL6-Timp-3−/−, uPA-NOG, B6J, ICR, Bash2, and BALB/C, with various degrees of prevalence. The pathogenicity of the murine astrovirus was not known.

Legislation in research

United Kingdom

In the U.K., as with all other vertebrates and some invertebrates, any scientific procedure which is likely to cause "pain, suffering, distress or lasting harm" is regulated by the Home Office under the Animals (Scientific Procedures) Act 1986. U.K. regulations are considered amongst the most comprehensive and rigorous in the world.[68] Detailed data on the use of laboratory mice (and other species) in research in the U.K. are published each year.[69] In the U.K. in 2013, there were a total of 3,077,115 regulated procedures on mice in scientific procedure establishments, licensed under the Act.[70]

United States

In the U.S., laboratory mice are not regulated under the

Association for Assessment and Accreditation of Laboratory Animal Care, which maintains the standards of care found within The Guide for the Care and Use of Laboratory Animals and the PHS policy. This accreditation is, however, not a prerequisite for federal funding, unlike the actual compliance.[71]

Limitations

While mice are by far the most widely used animals in biomedical research, recent studies have highlighted their limitations.

ALS,[77][78][79] Alzheimer's disease,[80] diabetes,[81][82] cancer,[83][84][85][86][87] multiple sclerosis,[88] Parkinson's disease,[88] and other illnesses has been called into question by a number of researchers. Regarding experiments on mice, some researchers have complained that "years and billions of dollars have been wasted following false leads" as a result of a preoccupation with the use of these animals in studies.[72]

Mice differ from humans in several immune properties: mice are more resistant to some

outbred, and nonlinear mice.[73]

An article in The Scientist notes, "The difficulties associated with using animal models for human disease result from the metabolic, anatomic, and cellular differences between humans and other creatures, but the problems go even deeper than that" including issues with the design and execution of the tests themselves.[76] In addition, the caging of laboratory animals may render them irrelevant models of human health because these animals lack day-to-day variations in experiences, agency, and challenges that they can overcome.[89] The impoverished environments inside small mouse cages can have deleterious influences on biomedical results, especially with respect to studies of mental health and of systems that depend upon healthy psychological states.[90]

For example, researchers have found that many mice in laboratories are obese from excess food and minimal exercise, which alters their physiology and drug metabolism.[91] Many laboratory animals, including mice, are chronically stressed, which can also negatively affect research outcomes and the ability to accurately extrapolate findings to humans.[92][93] Researchers have also noted that many studies involving mice are poorly designed, leading to questionable findings.[76][78][79]

Some studies suggests that inadequate published data in animal testing may result in irreproducible research, with missing details about how experiments are done are omitted from published papers or differences in testing that may introduce bias. Examples of hidden bias include a 2014 study from McGill University which suggests that mice handled by men rather than women showed higher stress levels.[94][5][95][96] Another study in 2016 suggested that gut microbiomes in mice may have an impact upon scientific research.[97]

Market size

The worldwide market for gene-altered mice is predicted to grow to $1.59 billion by 2022, growing at a rate of 7.5 percent per year.[98]

See also

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Further reading

External links

Wikimedia Commons has media related to Lab mice.
Wikispecies has information related to Mus musculus.

Taxonomy

Genetics

Media

Further reading

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