MICE

Scientific classification:

Kingdom: Animalia

Phylum: Chordata

Class: Mammalia

Order: Rodentia

Superfamily: Muroidea

Family: Muridae

Subfamily: Murinae

Genus: Mus

House Mouse: Mus musculus

The house mouse (Mus musculus) is a small rodent, a mouse, one of the most numerous species of
the genus Mus.

As a wild animal the house mouse mainly lives associated with humans, causing damage to crops and
stored food.

The house mouse has been domesticated as the pet or fancy mouse, and as the laboratory mouse which
is one of the most important model organisms in biology and medicine. It is by far the most commonly
used genetically altered laboratory mammal.

CHARACTERISTICS

House mice have an adult body length (nose to base of tail) of 7.5–10 cm (3.0–3.9 in) and a tail length of
5–10 cm (2.0–3.9 in). The weight is typically 10–25 g (0.4–0.9 oz). They vary in colour from white to grey,

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and light brown to black. They have short hair and a light belly. The ears and tail have little hair. The hind
feet are 15–19 mm (0.59–0.75 in) long; the normal gait is a run with a stride of about 4.5 cm (1.8 in),
though they can jump up to 45 cm (18 in). The droppings are blackish, about 3 mm (0.12 in) long, and
have a strong musty smell. The voice is a high-pitched squeak.

House mice thrive under a variety of conditions: they are found in and around homes and commercial
structures as well as in open fields and agricultural lands. House mice consume and contaminate food
meant for humans, pets, livestock, or other animals. In addition, they often cause considerable damage to
structures and property. They can transmit pathogens that cause diseases such as salmonellosis, a form
of food poisoning.

Young males and females are not easily distinguished: females have a significantly smaller distance
between their anus and genital opening. Females have 5 pairs of mammary glands and nipples; males
have no nipples. When sexually mature the most striking and obvious difference is the presence
of testicles on the males. These are large compared to the rest of the body and can be retracted into the
body. In addition to the regular pea-size thymus organ in the chest, house mice have a second functional
pinhead-size thymus organ in the neck next to the trachea.

BEHAVIOR

House mice usually run, walk or stand on all fours; but when eating, fighting or orienting themselves, they
stand only on the hind legs, supported by the tail. When running the horizontal tail serves for balance; the
end stands up vertically, unless the mouse is frightened. Mice are good jumpers, climbers, and
swimmers.

Mice are mostly active during dusk or night; they do not like bright lights. They have an instinctual fear of
so-called "black lighting" and strobe lighting, which leads to a common method of controlling mice in the
home. They live in a wide variety of hidden places that are near food sources and construct nests from
various soft materials. Mice are territorial and one dominant male usually lives together with several
females and young. Dominant males respect each other's territory and normally enter another's territory
only if it is vacant. If two or more males are held together in a cage, they will often turn aggressive unless
they have been raised together from birth.

House mice primarily feed on plant matter, but they will also accept meat and dairy products. Although
they are generally known to like fruits, they are repelled by the scent of many varieties of artificial fruit

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scent, for example strawberry or vanilla-scented candles. The reason for this is unknown, although it
dates back to antiquity when Roman Senators used candles scented with strawberry oils to keep mice out
of their sleeping chambers. They will drink water but require little of it, relying mainly on the moisture
present in their food. They will eat their droppings to acquire nutrients produced by bacteria in their
intestines. House mice, like other rodents, do not vomit.

Mice are afraid of rats, which often kill and (partially) eat them. This rat behaviour is known as muricide .
Despite this behaviour free-living populations of rats and mice do exist together in forest areas in New
Zealand, North America and elsewhere. House mice are generally poor competitors and in most areas
cannot survive away from human settlements in areas where other small mammals, such as wood mice,
are present. However in some areas (such as Australia) mice are able to co-exist with other small rodent
species.

Senses and communication

As primarily nocturnal animals, house mice have little or no colour vision. They have a sharp sense of
hearing and can perceive ultrasound, possibly up to 100 kHz. They communicate both in the human
audible range with squeaks (for long-distance warnings), and in the ultrasound range (for short-distance
communication).

House mice also rely on pheromones for social communication, some of which are produced by
the preputial glands of both sexes. The tear fluid and urine of male mice also contains pheromones, such
as major urinary proteins. Mice detect pheromones mainly with the vomeronasal organ (Jacobson's
organ), located at the bottom of the nose.

The urine of house mice, especially which of males, has a characteristic strong odor. At least ten different
compounds such as alkanes, alcohols, etc. are detectable in the urine. Among the ten, five compounds
are specific to males, namely 3-cyclohexene-1-methanol, Aminotriazole (3-amino-s-triazole), 4-ethyl
phenol, 3-ethyl-2,7-dimethyl octane and 1-iodoundecane.

Mice can sense surfaces and air movements with their whiskers.

Laboratory Mice

Mice are the most commonly used animal research model with hundreds of established inbred, outbred,
and transgenic strains. In the United States, they are not regulated under the Animal Welfare Act (AWA)

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administered by the USDA APHIS. However, the Public Health Service Act(PHS) as administered by
the National Institutes of Health (NIH) does offer a standard for care and use. Compliance with PHS is
required in order to receive federal funding. PHS Policy is administered by the Office of Laboratory
Animal Welfare (OLAW). Many academic research institutes seek accreditation voluntarily, often
through Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC), 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 voluntary not a prerequisite for federal funding.

Mice are common experimental animals in biology and psychology; primarily because they are mammals,
are relatively easy to maintain and handle, reproduce quickly, and share a high degree of homology with
humans. The mouse genome has been sequenced, and many mouse genes have human homologues.

In addition to being small, relatively inexpensive, and easily maintained, there are further benefits to the
use of mice in laboratory research. Because mice can reproduce quickly, several generations of mice can
be observed in a relatively short period of time.

Most laboratory mice are hybrids of different subspecies, most commonly of Mus musculus
domesticus and Mus musculus musculus. Laboratory mice come in a variety of coat colours including
agouti, black and albino. Many (but not all) laboratory strains are inbred, so as to make them genetically
almost identical. The different strains are identified with specific letter-digit combinations; for
example C57BL/6 and BALB/c.

The first such inbred strains were produced by Clarence Cook Little in 1909. Little was influential in
promoting the mouse as a laboratory organism.

Mutant and transgenic strains

Various mutant strains of mice have been created by a number of methods:

 Mice resulting from ordinary breeding.

 NOD mice, which develop diabetes mellitus type 1.
 MRL mice, mice with unusual regenerative capacities.
 "Waltzing" mice, which walk in a circular pattern due to a mutation adversely affecting their
inner ear.

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  Immunodeficient nude mice, lacking hair and a thymus. The mice don't produce T
lymphocytes and therefore don't mount cellular immune responses. They are used for research
in immunology and transplantation.
 Severe Combined Immunodeficient or SCID mice, with an almost completely defective immune
system.
 Transgenic mice, with foreign genes inserted into their genome.
 Abnormally large mice, with an inserted rat growth hormone gene.
 Oncomice, with an activated oncogene, so as to significantly increase the incidence of cancer.
 Doogie mice, with enhanced NMDA receptor function, resulting in improved memory and
learning.
 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.

Breeding

Mice have a rapid reproduction rate; the gestation period is approximately 19 to 23 days. The typical
litter size is 4 to 12 young. In some instances, up to thirty young have been born. Males can mate with the
female as soon as the litter is born, which means that a female could become pregnant with another litter
while still nursing her first, which results in less milk to go around and smaller, weaker pups. Female mice
should not be bred before 12 weeks or after 8 months; doing so can be very dangerous, and some mice
can die while giving birth. Females come into heat around every three to five days, so the pair can be
kept together for up to ten days. Baby mice, called pinkies or pups, are born blind, naked, and deaf. Their
eyes are closed and their ears are stuck to the sides of their heads. Mothers may eat any dead or sickly
offspring. Pups begin to grow hair at two to four days. Ears open at three to five days, and the pups will
start vocalizing. Eyes open at 14 days, and the babies will start exploring the world around them. At three
weeks old they look like miniature versions of adult mice. At four weeks the males in the litter should be
separated out so as to not cause the mother and sisters to become pregnant, the females can be left in
with the mother.

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 MOUSE FACTS

 Average litter size: 6-8 pups

 Gestation period: 19-21 days
 Estrus cycle: 4-5 days
 Postpartum estrus: females can become pregnant within 24 hours of giving birth, but
embryos conceived during postpartum estrus will not develop at the usual rate due to
delayed implantation
 Age at sexual maturity: 5-8 weeks (males tend to mature later than females)
 Minimum weaning age: about 19-21 days (can be done sooner if softened food is placed in
the bottom of the cage)
 Genome size: about 2.6 x 10^9 bp
 Normal diploid chromosome number: 40
 Post-natal development (adapted from the Jackson Laboratory):
o 0 Day Old: Blood red; possible milk spot.
o 1 Day Old: Lighter color red; milk spot present.
o 2 Day Old: Ears appear as nubs; pigment may start to appear in some strains.
o 3 Day Old: Ear flap starting to come away from head (one or both).
o 4 Day Old: Ears fully developed; completely off head; ears may start to migrate
towards back.
o 5 Day Old: Ears all back; skin appears much thicker with more color density to skin
o 6 Day Old: Milk spot disappearing or gone; skin still shows; colored fuzz appears
behind ears or on neck.
o 7 Day Old: Colored fuzz starts to cover pup; skin does not show.
o 8 Day Old: Belly starts to show fur; lower incisors show but not erupted.
o 9 Day Old: The fur is now thicker; females may start to show inguinal nipples (there
are five pairs of mammaries).
o 10 Day Old: Lower incisors erupted.
o 11 Day Old: Upper incisors erupted.
o 12 Day Old: Eyes still closed.
o 13 Day Old: Eyes open.

GUIDELINES FOR EFFICIENT MOUSE COLONY M AN AGEMENT

The goal of managing any mouse colony should be to maintain adequate numbers of animals in as little

shelf space as possible, while adhering to the policies regarding health and well-being of the mice, and

minimizing labor costs.

What constitutes “adequate numbers” will, of course, depend on a number of factors, including:

o experimental needs,
o breeding characteristics of a given strain,

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 o genotypes and phenotypes (morbidity/mortality) of individual mice,
o limits of approved animal use protocol.

Therefore, these guidelines will address in detail only the minimum effort and cage space needed to keep

a normal strain of mice “on the shelf”. However, the principles of mouse numbering, culling, and

replenishment can be utilized on any scale to minimize cage space.

Different strains of mice vary in fecundity, and certain mutant strains can be difficult to breed, due to a

variety of factors such as small litter sizes, low fertility, poor mothering instincts, high rates of cannibalism

of newborns, and higher morbidity or mortality resulting from the genetic mutation/alteration. When

acquiring a new genetically modified strain, therefore, it is always a good idea to consult with someone

who has direct experience with maintaining that strain.

 Cages required to maintain a strain: Simple strain maintenance generally requires no

more than 2-3 mating cages and 3-4 additional cages to hold weaned pups that will be used
to replace old breeders. To predict the cages needed to produce mice for experiments,
figure about one litter per month from young breeders, and about 6-8 pups/litter, until
experience proves otherwise. Maintaining these production levels requires consistent
replacement of older breeders.
 Breeding: House breeders in pairs of one male and one female. Gestation lasts 19-21
days. Check cages at least twice each week to flag pregnant females and record
approximate birth dates. Pups should be weaned (removed from breeding cage and
separated by sex) at 3 weeks of age. Identification can be accomplished prior to weaning by
any of several methods (see below). Tissue for genotyping is usually collected at the same
time that ID numbers are applied.
 Weaning: At 3 weeks of age, male and female pups are moved to separate holding cages,
with no more than 5 mice per cage. Females of any age or breeding status can be housed
together. To minimize fighting, weaned males should be group-housed only with their
littermates, and only until they are exposed to females. Males from different litters can be
grouped together only if they are no more than 4-5 days apart in age. Even littermates may
have to be separated as they age, to prevent fighting. Males that have been used as
breeders must ALWAYS be housed singly, because they will kill each other.
 Replacement of breeders: Replace a breeder pair if:
o they have not produced a litter in more than two months,
o they are producing small litters (1-3 pups per litter),
o they are killing their pups.
New breeder pairs should be 8-12 weeks of age. Although both males and females can breed up
to 8 months of age and beyond, if new breeders are available there is little point in continuing to
use breeders older than about 6 months. Also, a mouse that has not been allowed to breed prior to
about 4 months of age may never breed successfully. As breeders age, both litter size and the
frequency of litters decrease. It may be wise to wait for new breeders to produce a litter, before
euthanizing old breeders, if the strain presents any unusual breeding problems.

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  Cull older progeny: Euthanize older offspring as new litters are weaned. It is easy to
neglect weaned pups, since they are not breeding. Periodic evaluation of all cages is
essential to minimizing cage usage. If progeny are more than 3 months old, and you have
new litters to replace them, and you don’t need them for experiments, then take them off the
shelf. The only reason to keep more than one or two weaned litters for a strain that is not
being used in experiments is if that strain exhibits unusual morbidity or mortality, or is
otherwise difficult to breed.
 Pup identification: Weaned pups from different litters may be housed together if they are
uniquely identified (or if they don’t need to be identified). This helps to minimize the number
of holding cages. (See above, however, for caveats about males.) The following table lists
the common methods of pup ID, with pros and cons of each.

Method Pros Cons

Ear punching/ Simple, inexpensive, easy to read Sometimes ambiguous, subject to tearing and
notching healing, limited numbers

Metal ear tags Unique numbering, relatively inexpensive Loss of tags, infections, hard to read

Toe clipping Simple, inexpensive, permanent Less humane, must be done at an early age

Tattooing Relatively permanent, easy to read, may More difficult and time-consuming, may fade
be done on newborns with time if done improperly

Microchips Permanent, virtually unlimited numbers, High cost per mouse, difficult to apply,
can provide physiologic data requires expensive reader

AN EX AM PL E O F LOW -L EVEL M AIN T EN AN C E

Week 1:
Mating Mating
Holding Holding Holding Holding
cage 1 cage 2
cage cage cage cage

7 pups 5 pups 4 males 5 females 3 males 4 males

3 wks old 2 wks old 15 wks old 15 wks old 10 wks old 8 wks old

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The progeny from mating cage 1 are ready to be weaned. Three females and four males are found. They

are numbered and placed in two cages. The two cages of older progeny are discarded (shaded boxes).

Result: no change in cage number.

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Week 2:
Mating Mating
Holding Holding Holding Holding
cage 1 cage 2
cage cage cage cage

8 pups 5 pups 3 males 4 males 4 males 3 females

newborn 3 wks old 11 wks old 9 wks old 4 wks old 4 wks old

The progeny from mating cage 2 are ready to be weaned. Two females and three males are found and

numbered. The females are added to the cage of 3 females created in week 1, while the males are put

into a new cage. The oldest cage of male progeny is discarded (shaded). Result: no change in cage

number.

Week 3:
Mat. Mat.
Holding Holding Holding Holding
cage 1 cage 2
cage cage cage cage

8 pups No pups 4 males 4 males 5 females 3 males

1 wk old 9 wks old 5 wks old 4&5 wks old 4 wks old

The oldest male progeny can be discarded (shaded). No other action is necessary. Result: cage number

reduced to five.

DOS AND DON'TS OF MOUSE BREEDING

 Do minimize the amount of noise and traffic in your mouse room. Try to always have the
same person(s) take care of your mice. Ultrasonic noise, in particular, can disrupt normal
mouse behavior (including mating behavior), and can be produced by something as
innocuous as a dripping faucet.
 Do get to know your husbandry staff and maintain a good working relationship with them.
They will usually be the first people to notice when something is wrong with your mice.
 Do make use of the box at the bottom of the standard cage card to identify the mice in that
cage
 Don't bang cages around or jostle the mice any more than necessary. Handle them calmly
and quietly.

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 Do allow male mice a little time (e.g., a day) to get used to a new cage before expecting
them to mate successfully. Males like to mark their territory before "getting down to
business".
 Don't let mice get too old prior to first mating, if you are planning to breed them. It's best to
give them at least some practise within 4 months of birth.
 Don't expect breeding females to be productive after more than 8 months of age, although
they can have litters well beyond that. Males are generally productive at least a year, and
often well beyond that. Maximize production of pups by replacing your breeders with
younger mice on a regular schedule.
 Do keep records of breeding performance so time and resources aren't wasted on
unproductive mice. Not all mice will breed successfully, even among standard strains.
 Do cycle females through a male's cage to maximize production from a small number of
males. The estrus cycle is 4-5 days long, so females should be left in the cage at least this
long, unless you are checking for plugs.
 Don't cycle males through a female's cage - pheromones from new males can disrupt
pregnancies initiated by previous males.
 Don't disturb gestating females any more than necessary, especially a few days before and
after birth.
 Do use "Do Not Change" cards on the cages of gestating females to minimize disturbances
a few days before and after birth. If a cage needs changing while a Do Not Change card is
on it, you will be responsible for changing the cage.
 Do leave a female with the same male continuously to take advantage of postpartum estrus.
The female can get pregnant within 24 hours of giving birth and normal males will not harm
their pups. If you have plenty of males, 1:1 matings are recommended to maximize
production while minimizing labor.
 Do use dirty bedding from a male's cage to stimulate estrus in females. Females group-
housed for long periods can go into anestrus, i.e., they will stop cycling normally. This is
reversed by exposure to male pheromones present in the urine.
 Do check for vaginal plugs if you want to know exactly when a female has mated
successfully. This is best done in the morning because the plugs will start to fall out after a
few hours. The presence of a plug is not absolute proof of pregnancy, but it is a very reliable
indicator. On the other hand, the absence of a plug is not a guarantee that the female isn't
pregnant.
 Do communicate with the vivarium manager and husbandry staff about any special
precautions for your mice, e.g., autoclaved water, cages, and bedding, and irradiated (or
autoclaved) feed for immunocompromised mice. Special diets are available and those that
aren't can be custom made by the manufacturer.
 Do give your mice some environmental enrichment. Nestlets are an inexpensive choice -
they are compressed cotton squares that mice love to tear apart and fluff up into a warm
nest. Other options include plastic or cardboard tubes, or "shelters". Nesting material of
some sort should be given to all breeding females.
 Do keep your mice on a regular light/dark cycle with at least 12 hours of light. The TMF uses
a 14-hour light cycle. Periodically check that the lights actually come on and go off as
scheduled. Avoid entering mouse rooms during dark periods. A red light can be used to
minimize impact on the mice if you have to work in the room during dark hours.

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 CROSS-FOSTERING OF MOUSE PUPS

Many factors affect the survival of newborn mice, including the mother’s nurturing instincts, the number of

pups in the litter (too many and too few are bad), amount of milk production, tendency for cannibalism,

etc. Some problems can be overcome by moving the pups to the cage of another lactating female that will

take better care of them. This is known as cross-fostering. Our protocol for cross-fostering is as follows:
1. Donor and recipient litters should be within 2 days of age, and neither litter should be more
than 4 days of age, for best results.
2. If possible, use a recipient strain with a different coat color from the donor strain, for easier
identification of the cross-fostered pups. The recipient strain is normally an outbred strain
such as CD-1, ICR, or Swiss, because these tend to make the best moms.
3. Remove the recipient mom from her cage. When you pick her up, try to get her to urinate
on your gloves, and wipe the urine on the pups you are transferring. (Most mice will urinate
if turned upside down.)
4. Remove some (or most) of the recipient’s pups if she has a large litter. After cross-
fostering, the total number of pups in her cage should be 5-10.
5. Mingle the transferred pups with the recipient’s pups and rub them with dirty bedding to
give them the same scent as the recipient’s pups. Add nesting material if necessary.
6. Return the recipient mom to her cage and place it back on the rack.
7. Observe the recipient mom’s behavior after a few minutes. If she settles down to nurse the
pups, or is grooming them, she will probably accept the transferred pups. If she scatters
them around the cage, it is unlikely she will take care of them.

PHEROMONE EFFECTS

Mouse reproductive behavior is governed to a large extent by pheromones. Three important effects of

pheromones are described here.

Lee-Boot Effect

Housing female mice in groups will result in synchronization of their estrus cycles. Prolonged absence of

male pheromones results in a state of anestrus (lack of a normal estrus cycle).

Whitten Effect

Estrus can be induced in most group-housed females by adding male mouse urine (or dirty bedding from

a male’s cage) to their cage.

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Bruce Effect

Pheromones from a strange male can prevent the implantation of embryos into the uterine walls of a

recently bred female. This is why one should not move a female from one male’s cage to another.

The Lee-Boot and Whitten Effects can be utilized to produce closely synchronized pregnancies. After

group-housing females separately from males for some time, put them in a male’s cage. Generally at

least 75% of the females will become pregnant within 3 days.

Males prefer to have their territory scent-marked before they will breed efficiently. Therefore, it is better to

leave males in their “home” cage and shuffle females in and out to maximize the production from a small

number of males.

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