UNC-Institutional Animal Care and Use Committee

Mouse Colony Management Lecture

Emily Weston, MS, rLAT

Training and Compliance Manager,

Office of Animal Care and Use

February 2018 IACUC UNC-CH 1

This tutorial is designed to provide basic information on maintaining a breeding

colony

Included topics:

• Mouse life cycle & reproduction basics

• Colony management: documentation & strategies

• Other: Health issues, identification, compliance, and resources

February 2018 IACUC UNC-CH 2

Basics of Mouse Reproduction

February 2018 IACUC UNC-CH 3

• Sexual maturity: • Males ~6 wks• Females ~8 wks

• If bred early, generally produce small/weak litters

• Estrous cycle—4 to 5 days (Estrus ~10 hours)

• Ovulation—2 to 3 hours post estrus onset

• Gestation—18 to 21 days

• Breeding life—8 months to 1 year

Breeding Performance Factors

Hybrid Vigor

Hybrids stronger than inbred strains usually

Mutations/Transgenes

Temperature/Humidity

Stable and within range

Light cycle/intensity

Nocturnal

Consistent and uninterrupted

Health Status For more details:

http://vetbiotech.com/resources.php?id=76&site_cat=50

Noise & Vibration

Construction, radios

Odors

Noxious, perfumes

Handling

Breeders, new litters

Nutrition

Fat content

Enrichment

Nestlets

February 2018IACUC UNC-CH 4

Other Factors to Consider

Replace breeders before reproductive performance declines

Replace non-productive breeders (~60 days of mating/last litter)

Mate mice early in maturity

Use experienced males with young females

Keep detailed breeding records

February 2018 IACUC UNC-CH 5

Strain Dependent

Mouse strains vary!

Check your strain norms!!!

IACUC UNC-CH 7

Differentiating Sexes

Males have greater anogenital distance

http://www.lawte.org/materials/nirah/weaning_mice.ppt

February 2018

MALE (black dot) FEMALE

IACUC UNC-CH 8

Monitoring Estrus Vulvar swelling

Cell culture

Copulatory plugs

Check early in the morning, breeding usually occurs at during dark cycle

Plugged Female

February 2018

https://www.jax.org/news-and-insights/jax-blog/2014/september/six-steps-for-setting-up-timed-pregnant-mice

IACUC UNC-CH 9

Postpartum Estrus

Females come into estrus within 24 hours of bearing litter

Can be impregnated if male is present

Ideally first litter will be ready to wean when second litter is due

February 2018

IACUC UNC-CH 10

Breeding Strategies

Continuous pair mating Remember postpartum estrus!

Female communal nesting non-pregnant females

Single mother (remove male) -if large litters, helps with cage density

Continuous trio breeding -very intensive

ONLY one male per breeder cage!!

February 2018

IACUC UNC-CH 11

Intensive vs Nonintensive

Intensive or continuous breeding Pairs together throughout

pregnancy and nursing

Labor intensive, maximizes breeding productivity

Nonintensive breeding Male housed separately while

female is pregnant or nursing

Allows recuperation period between pregnancies

February 2018

Whitten Effect: • Females housed together

10-14d

• Estrus suppressed

• Introduce male or androgenous odor to cage to resume estrus

• 4-5 day cycle, all ovulate on 3rd day

• Six Steps for Setting Up Timed Pregnancies- JAX

February 2018 IACUC UNC-CH 12

IACUC UNC-CH 13

DAY EVENT DAY EVENT

1 SKIN REAL RED;

SHORT, CURLY

WHISK.; UMB.CORDS

8 FUZZ ON BELLY

2 STRAIGHT, FINE

WHISKS.; SKIN

LIGHTER RED

9 FUR BACK; LAST DAY

BELLY TRANSLUCENT

,MILKBANDS

3 EAR “NUBS”

PROMINENT

10 TOP TEETH ERUPTED;

BELLY SLIGHT FUZZ

4 PINK SMOOTHER

SKIN; EAR FLAPS 1/2

UP, DOWN

11 FULLY FURRED; FUR

SHORT & SMOOTH

5 EARS UP; WHITE

FUZZ ACROSS

SHOULDERS

12 NIPPLES VISIBLE

6 BOTTOM TEETH

ERUPTED

13 EYES OPEN

7 ENTIRE BACK

SLIGHTLY FURRED

14 EARS OPEN

So now you’ve got some pups… A Quick Guide for Aging Neonates

February 2018

• JAX posters available upon request and placed in most DCM facilities

• DOB must be on cage w/in 3 days!

• This chart can be found on the back of the hot pink cards in DCM!

IACUC UNC-CH 14

Fostering

When all else fails…

Mother is not lactating (no milk spots)

Pups are scattered

Mother has died

….It’s time to try to foster

February 2018

IACUC UNC-CH 15

Tips for a Successful Foster

Ideally you will chose an Experienced foster with:

pups born within 48 hours of foster litter

Different coat colors, or ID pups

Keep the litter size same (reduce foster’s litter)

Urine from foster nest/litter helps w/ acceptance

Then mix the litters together in the foster’s nest

Leave them ALONE!

Foster mom should begin nursing/grooming within 6 hours

February 2018

IACUC UNC-CH 16

Strains vs Stocks Strains have little genetic variability

Inbreeding for no less than 20 generations

Stocks maximize genetic variability

Variability creates hardy mice

February 2018

IACUC UNC-CH 17

NomenclatureBALB/c StCrl p GF (F83)

Strains indicated by all caps and/or numbers

Substrains separated by diagonal (/)

Subline derivation (eg: preserved) indicated by lower case

Hygenic state (eg: Germ Free) may be included

Inbreeding level – F + number of brother/sister inbred generations

More info – www.informatics.jax.org/nomen

February 2018

RECORD KEEPING

??????

HOW DO I KEEP TRACK OF ALL THESE ANIMALS?

IACUC UNC-CH 19

P.I.

DCM

IACUC

REQUIREMENTS

February 2018

IACUC UNC-CH 20

IACUC requirements for Cage Cards

PI Name

Birthdate of pups if breeding cage (required within 3 days of birth)

Pre-printed cage card with IACUC protocol number (DCM RFID card)

Note: temporary cards allowed, but must include all above, plus date the RFID card was ordered

February 2018

IACUC UNC-CH 21

For Stock Cages, PI cards Should Include:

Mouse strain name; researcher

Genotype

Date of birth

Number of mice in cage

Mouse identification numbers

Sex

February 2018

IACUC UNC-CH 22

Example Stock cards for Weaned Animals

P.I room Cage #

ID # DOB. coat L #

Dam ID & genotype Sire ID & genotype

genotype commentsW.D.

F or M

February 2018

IACUC UNC-CH 23

For Breeding Cages, PI cards Should Include:

Strain & genotype

Researcher & room #

I.D. Number of each animal

Date cage set up/paired

For each litter of pups:

Number born

Date of birth

Litter number

Weaning date

February 2018

IACUC UNC-CH 24

Example Breeding Cage Card:

PI room Cage #

Dam ID #, DOB., L#, strain/genotype

Sire ID #, DOB., L#, strain/genotype

Set-up date, End date

Date out

Date out

_______________________________________

_______________________________________

_______________________________________

_______________________________________

preg # pups DOB. L# genotype commentsdam

February 2018

IACUC UNC-CH 25February 2018

IACUC UNC-CH 26

Other information

Phenotype

Experiment

Date of death

Reason for death

February 2018

IACUC UNC-CH 27

Record keeping methods

Paper/pen in facility, update online later

Computer/tablet

Databases – many software options available for free/purchase

Excel- can be difficult with multiple users

Keep database current

February 2018

IACUC UNC-CH 28

Identification Methods

See IACUC Standard

Ear Tag

Ear Notch

Toe Clip

Microchip

Tattoo

February 2018

IACUC UNC-CH 29

Tagging

Easy to administer

Tags can be lost

May cause irritation or loss of ear flap

May need to restrain mouse to read tag #

Mouse Techniques Packet, see pg. 13-14.

February 2018

IACUC UNC-CH 30

Ear Notch

Easy to read

Inexpensive

Can use tissue for genotyping

Some strains of mice are known for ear mutilation

Mouse Techniques Packet, see pg. 12.

February 2018

IACUC UNC-CH 31

Toe Clip

Permanent Identification Method

Early age identification

Requires scientific justification

Requires IACUC approval

Must pick up mice to read

Toe at 7 to 10 days old!

No more than 2 toes per foot, nor 4 toes per animal

February 2018

IACUC UNC-CH 32

Microchip ID

Permanent Identification Method

Easy to read

Some systems are programmable and provide telemetry

Expensive

Possible risk of infection (low)

February 2018

IACUC UNC-CH 33

Tatooing

Low health risk

Versatile

Somewhat difficult to administer

Requires skill, equipment $$

Ink fades over time

February 2018

IACUC UNC-CH 34

Additional Resources

February 2018

IACUC UNC-CH 35

MGI Glossary

Mouse Genome Informatics http://www.informatics.jax.org/

MGI is the international database resource for the laboratory mouse, providing integrated genetic,

genomic, and biological data to facilitate the study of human health and disease.

February 2018

Cryopreservation

UNC has its own Mutant Mouse Regional Resource Center (MMRRC)

To save space and save your mouse lines,

Contact the MMRRC—919-966-4570 or

mmrrc@med.unc.edu

IACUC UNC-CH 37

For Additional Help @ UNC

UNC School of Medicine Animal Models Core –Transgenic Mouse Line Production validation and use of CRISPR/Cas9 custom nucleases for

production of mutant mice, rats or cell lines

IACUC Training and Compliance Team– policies, standards, guidelines

DCM Vet Services

DCM Colony Management Services

February 2018

IACUC UNC-CH 38

Where to Get More Information

www.jax.org

www.criver.com

www.aalas.org

www.med.unc.edu/mmrrc/

www.research.unc.edu/iacuc/

www.research.unc.edu/DCM/

February 2018

Thank You!!