Toxicologic Pathology

 

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Toxicologic Pathology, Vol. 34, No. 6, 795-801 (2006)
DOI: 10.1080/01926230601009527

Articles

Optimal Sampling of Rat Liver Tissue for Toxicogenomic Studies

Julie F. Foley1, Jennifer B. Collins2, David M. Umbach3, Sherry Grissom2, Gary A. Boorman4 and Alexandra N. Heinloth2

1 Laboratory of Experimental Pathology
2 National Center for Toxicogenomics
3 Biostatistics Branch and
4 Environmental Toxicology Program, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709, USA

Correspondence: Address correspondence to: Julie F. Foley, NIEHS, 111 T. W. Alexander Dr., Research Triangle Park, North Carolina 27709-2233, USA; e-mail:foley1{at}niehs.nih.gov

Different degrees of a toxic response between and within the various lobes of the liver have been observed in rodents following treatment with acetaminophen. This study was designed to compare 2 sampling methods of the rat liver for gene-expression analysis. Ten male Fischer 344/N rats, 12–14 weeks of age, were treated with vehicle (0.5% aqueous ethyl cellulose) or acetaminophen (APAP, 1500 mg/kg) and sacrificed 24 hours following dose administration. Two representative sections were collected from the left liver lobe, stained with hematoxylin and eosin (H&E), and evaluated independently by 2 pathologists. The central core of the left lobe was cubed and frozen. Five random cubes were conserved, while the remaining left lobe core was pulverized. From each of the 10 animals, 2 random cubes and 2 samples from the homogeneous, pulverized samples were prepared for microarray analysis. Histopathologic evaluation revealed a variable response of centrilobular necrosis within the left lobe. Multiple methods used to analyze the microarray data indicated that sampling technique was not a major contributor to the variability observed in the gene expression data; however, only the powdered samples clustered for all animals, even those with disparate histopathologic results. Additionally, a powdered sample provided the advantages of a homogenous sample pool and the ability to use sample aliquots for other analyses to include proteomics, metabonomics, and other molecular techniques.

Key Words: Liver • acetaminophen • histology • pathology • microarray • rat • necrosis

Abbreviations: APAP, acetaminophen • C, cube • CV, central vein • H&E, hematoxylin and eosin • LN2, liquid nitrogen • NBF, 10% neutral-buffered formalin • P, powder • PC, principal component • PCA, principal component analysis


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