Mouse Colony Management Lecture

Transcription

1 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 IACUC UNC-CH 1

2 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 IACUC UNC-CH 2

3 Basics of Mouse Reproduction 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 IACUC UNC-CH 3

4 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: d=76&site_cat=50 Noise & Vibration Construction, radios Odors Noxious, perfumes Handling Breeders, new litters Nutrition Fat content Enrichment Nestlets IACUC UNC-CH 4

5 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 IACUC UNC-CH 5

6 Strain Dependent Mouse strains vary! Check your strain norms!!!

7 Differentiating Sexes Males have greater anogenital distance materials/nirah/weanin g_mice.ppt MALE (black dot) FEMALE IACUC UNC-CH 7

8 Vulvar swelling Cell culture Copulatory plugs Check early in the morning, breeding usually occurs at during dark cycle Monitoring Estrus Plugged Female IACUC UNC-CH 8

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 IACUC UNC-CH 9

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!! IACUC UNC-CH 10

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 IACUC UNC-CH 11

12 Females housed together 10-14d Estrus suppressed Introduce male or androgenous odor to cage to resume estrus 4-5 day cycle, all ovulate on 3 rd day Six Steps for Setting Up Timed Pregnancies- JAX Whitten Effect: IACUC UNC-CH 12

13 So now you ve got some pups A Quick Guide for Aging Neonates DAY EVENT 1 SKIN REAL RED; SHORT, CURLY WHISK.; UMB.CORDS 2 STRAIGHT, FINE WHISKS.; SKIN LIGHTER RED 3 EAR NUBS PROMINENT 4 PINK SMOOTHER SKIN; EAR FLAPS 1/2 UP, DOWN 5 EARS UP; WHITE FUZZ ACROSS SHOULDERS 6 BOTTOM TEETH ERUPTED 7 ENTIRE BACK SLIGHTLY FURRED DAY EVENT 8 FUZZ ON BELLY 9 FUR BACK; LAST DAY BELLY TRANSLUCENT,MILKBANDS 10 TOP TEETH ERUPTED; BELLY SLIGHT FUZZ 11 FULLY FURRED; FUR SHORT & SMOOTH 12 NIPPLES VISIBLE 13 EYES OPEN 14 EARS OPEN 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 13

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 IACUC UNC-CH 14

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 IACUC UNC-CH 15

16 Strains vs Stocks Strains have little genetic variability Inbreeding for no less than 20 generations Stocks maximize genetic variability Variability creates hardy mice IACUC UNC-CH 16

17 Nomenclature BALB/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 IACUC UNC-CH 17

18 RECORD KEEPING?????? HOW DO I KEEP TRACK OF ALL THESE ANIMALS?

19 P.I. DCM REQUIREMENTS IACUC IACUC UNC-CH 19

20 IACUC requirements for Cage PI Name Cards 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 IACUC UNC-CH 20

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 IACUC UNC-CH 21

22 Example Stock cards for Weaned Animals P.I room F or M Cage # Dam ID & genotype Sire ID & genotype ID # DOB. W.D. coat L # genotype comments IACUC UNC-CH 22

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 IACUC UNC-CH 23

24 Example Breeding Cage Card: PI room Cage # Dam ID #, DOB., L#, strain/genotypedate out Sire ID #, DOB., L#, strain/genotype Date out Set-up date, End date preg dam # pups DOB. L# genotype comments IACUC UNC-CH 24

25 IACUC UNC-CH 25

26 Other information Phenotype Experiment Date of death Reason for death IACUC UNC-CH 26

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 IACUC UNC-CH 27

28 Identification Methods See IACUC Standard Ear Tag Ear Notch Toe Clip Microchip Tattoo IACUC UNC-CH 28

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 IACUC UNC-CH 29

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. IACUC UNC-CH 30

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 IACUC UNC-CH 31

32 Microchip ID Permanent Identification Method Easy to read Some systems are programmable and provide telemetry Expensive Possible risk of infection (low) IACUC UNC-CH 32

33 Tatooing Low health risk Versatile Somewhat difficult to administer Requires skill, equipment $$ Ink fades over time IACUC UNC-CH 33

34 Additional Resources IACUC UNC-CH 34

35 MGI Glossary Mouse Genome Informatics 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. IACUC UNC-CH 35

36 Cryopreservation UNC has its own Mutant Mouse Regional Resource Center (MMRRC) To save space and save your mouse lines, Contact the MMRRC or

37 For Additional 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 IACUC UNC-CH 37

38 Where to Get More Information IACUC UNC-CH 38

39 Thank You!!