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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
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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
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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
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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
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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
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Strain Dependent
Mouse strains vary!
Check your strain norms!!!
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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
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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
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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
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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
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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
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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
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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!
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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
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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
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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
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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
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RECORD KEEPING
??????
HOW DO I KEEP TRACK OF ALL THESE ANIMALS?
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IACUC UNC-CH 19
P.I.
DCM
IACUC
REQUIREMENTS
February 2018
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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
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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
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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
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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
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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
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IACUC UNC-CH 25February 2018
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IACUC UNC-CH 26
Other information
Phenotype
Experiment
Date of death
Reason for death
February 2018
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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
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IACUC UNC-CH 28
Identification Methods
See IACUC Standard
Ear Tag
Ear Notch
Toe Clip
Microchip
Tattoo
February 2018
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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
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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
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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
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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
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IACUC UNC-CH 33
Tatooing
Low health risk
Versatile
Somewhat difficult to administer
Requires skill, equipment $$
Ink fades over time
February 2018
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IACUC UNC-CH 34
Additional Resources
February 2018
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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
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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
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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
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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
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Thank You!!