mouse pathology 8.3.2012 2 - unifr.ch · 2017. 2. 2. · mouse pathology 08.03.2012 andreas...

Mouse pathology 08.03.2012

Andreas Pospischil, IVPZ 1

Institute of Veterinary Pathology

08.03.12

Mouse pathology Andreas Pospischil


Post-mortem investigation mouse

• Why?

• How?

• Next steps?

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Post-mortem investigation in mice

Why? •  Progress in functional genomics of the mouse

•  Phenotyping of mutants helps understand the normal / mutant function of gene products

•  Phenotyping depends on histopathological comparison of mutant to non-mutant individuals and analysis of the lesion and pathology in mutant / non-mutant mouse individuals





Post-mortem investigation (necropsy) in mice

How? •  Evaluation and collection of all organs and lesions

•  Systematic, efficient and simple procedures (SOP -> OECD guidance document) ensure repro-ducibility in a large series of animals and by different prosectors

•  Body weight

•  organ weight of heart, liver, spleen, right and left kidney, testis / uterus, ovaries and prominent lesions (e.g. neoplasms)




Institute of Veterinary Pathology Institute of Veterinary Pathology





Post-mortem investigation (necropsy) in mice: Examples


Institute of Veterinary Pathology Institute of Veterinary Pathology












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Post-mortem investigation (necropsy) in mice: Examples, documentation of lesion:



Fixation of tissues

Formalin fixation 5 - 10% buffered (!!!!) formalin is the fixative of choice. Tissues can stay in formalin forever with good morphologic preservation.

However, prolonged fixation affects immunodetection. The following protocol is intended for optimal immunoreactivity preservation.

Fix small pieces of tissues for minimum 6h (at RT), maximum 48 h (at +4°C). OPTIMAL 8-10h.





Demo-video






Reproducibility (specimen collection, evaluation, -> SOP)





Preparation of formalin-fixed tissue samples



Preparation of formalin-fixed tissue samples



Tissue section





Reading of tissue sections

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Interpretation of histopathology slides

•  Next steps? •  Correct nomenclature of mice, genes, mutations

etc. (-> SOP) •  Reporting methods and terminology (-> SOP)

•  Reproducibility (specimen collection, evaluation, -> SOP)

•  Labeling and and documentation (-> SOP)

•  Background lesions of mouse strains used („normal pathology“)

•  Health and pathogen status of the colony (SPF)




Next steps?

Correct nomenclature of mice, genes, mutations etc. (-> SOP):

Guidelines for Nomenclature of Genes, Genetic Markers, Alleles, and Mutations in Mouse and Rat -> http://www.informatics.jax.org/mgihome/nomen/gene.shtml






§  Reporting methods and terminology (-> local SOP)

§  Reproducibility (specimen collection, evaluation, -> local SOP)

§  Labeling and and documentation (-> local SOP)



Mutant mouse pathology

European mutant mouse pathology database

http://www.pathbase.net/

http://eulep.pdn.cam.ac.uk/Pathology_Ontology/createOntologyPopUp%28mpath%29.php



Pathbase





Background lesions of mouse strains used („normal pathology“)



Health and pathogen status of the colony (SPF)

• Mouse hepatitis virus (MHV)

• Mouse parvovirus

• Murine Helicobacter species • Citrobacter freundii

• Mycoplasma pulmonis

•  Sendai virus



Health and pathogen status of the colony (SPF): Mouse hepatitis virus (MHV):

RNA virus enveloped Coronaviridae Common in wild and laboratory mice In general, in immunocomnpetent mice asymptomatic; disease depends on age, genotype, sanitary and experimental status and the visulence of the infecting virus strain

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Health and pathogen status of the colony (SPF): murine parvovirus

DNA virus, nonenveloped Parvoviridae

Wild and laboratory mice

No clinical signs, self limiting disease,

However, interfering with experimental design



Health and pathogen status of the colony (SPF): murine Helicobacter species

Helicobacter (H.) species (spp.) are Gram-negative, microaerophilic, spiral-to-curve-shaped bacteria with sheathed flagella.

They commonly colonize the GI tract. H. muridarum (Lee et al., 1992), "H. (Flexispira) rappini" (Schauer et al., 1993), H. hepaticus (Fox et al., 1994), and H. bilis (Fox et al., 1995) have been identified in the lower GI tracts of mice.

Diagnosis polymerase chain reaction (PCR),

bacteriologic culture,

histology


(PCR),

Page 41


Health and pathogen status of the colony (SPF): Citrobacter freundii

biotype 4280 (Citrobacter rodentium)

Non-motile, Gram-negative rod

Enterobacteriaceae

Disease allmost exclusively in weanling mice





Health and pathogen status of the colony (SPF): Mycoplasma pulmonis Small pleomorphic bacteria lacking a cell wall

Mycoplasmataceae Colonize middle ear and nasopharynx Clinical signs: dyspnoea, weight loss, ruff hair coat

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Health and pathogen status of the colony (SPF): Sendai virus

RNA virus, enveloped

Paramyxoviridae

Mice, rats, hamsters, guinea pigs In immunodeficient rodents: pneumonia




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