February 26, 2008

Mouse Reproduction,
Genetic Crosses,
Inbred and Congenic Strains
The Life Cycle of the Mouse

~70
Important inbred strains of Mus musculus

Mouse Genetics, Lee M. Silver, Oxford University Press, 1995.

Laboratory Mouse Suppliers
in the USA

•The Jackson Laboratory (JAX)

•Charles River Laboratory (CR)

•Taconic Farms (Taconic)
 March 9, 2004

Mouse Genetics, Lee M. Silver, Oxford University Press, 1995.

Reproductive performance:
Fecundity 生產力
Productive mating frequency X (litter size) X (number of litters)

Fecundity:
Female: reduced by the age of 8-10 months
Male: remain fertile through their lives. However,
older male that have become obese and sedentary
are unlikely to breed.

Puberty:
Female: first ovulate naturally between 6-8 weeks
3-6 weeks can be induced to ovulate
Male: onset of puberty occurs 34-38 days
 The Estrus Cycle of Female
1. Normal estrus cycle is 4-6 days in length.
2. Four phases of the estrus cycle:
a. Proestrus: a new bath of eggs reach maturity
b. Estrus: ovulation of fully mature oocytes (begin
after midnight and last for 6-8 hours)
c. Metestrus: mature eggs move through the
oviducts and into the uterus. (The act of
successful copulation induces hormonal
changes that prepare the uterus for pregnancy.)
d. Diestrus: unfertilized eggs are eliminated

Sterile copulation (no fertilization):

Induce a state of pseudopregnancy.
Plug:
1. After a successful copulation has been completed,
particular components of the male ejaculate (from
the vesicular and coagulating glands) will
coagulate to form a hard plug that occludes the
entrance to the vagina.

2. Plugging should be performed as early as

possible in the morning after a potential mating.
Usually persists for 16-24 hours after copulation.

Weaning:
Mice can be weaned at 18-28 days old.
Categories of Genetic Crosses

Mouse Genetics, Lee M. Silver, Oxford University Press, 1995.

Locus (gene): A, a

Male, A/A Male, a/a

Female, A/A Female, a/a

Male or female, A/A Male or female, a/a

Sex unknown Sex unknown

Incross

A/A A/A a/a a/a

 Outcross
A/A a/a

a/a A/a A/a A/a A/a A/A

Intercross

Backcross Backcross
a/a A/A A/a A/a

a/a a/a A/a A/a A/A A/A A/a A/a

The Generation of Inbred Strains
Inbreeding:
A process leading to the production of inbred mice
that are genetically homogeneous and homozygous
at all loci.

Inbred Strain:
The “International Committee on Standardized
Nomenclature for Mice” has ruled that a strain of
mice can be considered “inbred” at generation F20
of brother-sister matting.

F: filial generation
F1: first filial generation; F2: second filial generation
 The Generation of Inbred Strains

1. There is a 12.5% chance that

both F2 progenitors (F3) are
identically homozygous at any
one locus.

2. All offspring in all subsequent

filial generations will also be
homozygous for the same
alleles at these particular loci.

Figure 3.1 Consequences of inbreeding at the F2 generation.

Mouse Genetics, Lee M. Silver, Oxford University Press, 1995.
Operational Definition of Inbred Homozygous status
F20: 98.7%
F30: 99.8%
F40: 99.98%
F60: 100%

Mouse Genetics, Lee M. Silver, Oxford University Press, 1995.

Effects of genetic background
Normal development and physiology can vary significantly
from one strain of mice to the next. It is often not possible to
distinguish subtle effects due to the mutation itself from
genetic background.

Coisogenic Strain
The mutant animal differs at only a single locus from non-mutant
animals of the same strain. The new mutant strain is coisogenic
because its genome is identical (isogenic) to that of its sister
strain except at the mutant locus.
e.g. BALB/cJ-nu, mice with nude mutation of BALB/cJ strain
Congenic Strain
Mice that have been bred to be essentially isogenic with an inbred
strain except for a selected differential chromosomal segment.

Consomic Strain
Mice that have been bred to be essentially isogenic with an inbred
strain except for a selected whole chromosome.
 FVB/N B6

Congenic Strain
F1
50%

N2
75%

N3
87.5%

N4
93.75%

Mouse Genetics, Lee M. Silver, Oxford University Press, 1995.

 Backcross
F1 N2 N3 N4 N5 N6 N7 N9
50% 75% 87.5% 93.75% 96.875% 98.4375% 99.21875% 99.8%

Mouse Genetics, Lee M. Silver, Oxford University Press, 1995.

 TRADITIONAL SPEED CONGENIC
Donor Recipient Donor Recipient
Strain A Strain B Strain A Strain B

a/a* b/b % loci a/b a/a* b/b

100% F1 100%
b/a* b/b b/a* b/b

50% N2 <30%
b/a* b/b b/a* b/b

25% N3 <5%
b/a* b/b b/a* b/b

12.5% N4 <0.2%

b/a* b/b b/a* b/b

6.25% N5
b/b b/a*
b/a*
Inbred B congenic

MARKER-ASSISTED SELECTION
<0.2% N10
b/a* Average time to completion:
Inbred B congenic 5 generations X 3 months/generation
=> 15 months
Average time to completion:
10 generations X 3 months/generation
=> 30 months
Genesis 28: 164-166 (2000)

- ES cells are usually derived from 129Sv

strains
- Knockout mice are usually mixed strains of
C57BL/6 and 129Sv
- Screening with simple sequence length
polymorphism (SSLP)
- MIT/Whitehead database
- The 56 primer pairs spread ~ 30cM apart
- Regular agarose electrophoresis
 Volume 14: 350-356 (2003)

Mouse Informative Markers – Multiple Strains

http://www.well.ox.ac.uk/rat_mapping_resourses/
http://jaxmice.jax.org/services/speedcongenic.html
http://www.cidr.jhmi.edu/mouse/mult_inf.html
Normal Karyote of Mus musculus
2N=40

Mouse Genetics, Lee M. Silver, Oxford University Press, 1995.

 Normal Karyotype of Mus musculus
1. 20 pairs of chromosomes
19 autosomal pairs, XY sex chromosomes
2. Telocentric chromosomes
A centromere at one end and a telomere at the other

3. Individual chromosome can be distinguished by

banding patterns.
G-bands: Giemsa-stained bands (DNA condense
early, replicate late, relatively A:T rich)

R-bands: Reverse G-bands (DNA condense late,

replicate early, relatively G:C rich)
Mouse Genetics, Lee M. Silver, Oxford University Press, 1995.