Toxicologic Pathology

 

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Toxicologic Pathology, Vol. 30, No. 2, 295 (2002)
DOI: 10.1080/019262302753559669

Routine Immunohistochemistry to Simplify Ovarian Follicle Counts

Levan Muskhelishvili

Pathology Associates-A Charles River Company, Jefferson, AR

John R. Latendresse

Pathology Associates-A Charles River Company, Jefferson, AR

Thomas J. Bucci

Pathology Associates-A Charles River Company, Jefferson, AR

U.S. and internationally harmonized Health Effects Test Guidelines for Reproduction and Fertility Effects (USEPA OPPTS 870,3800, 1998: OECD No. 416 C(83)44, 1983) include enumeration of primordial and developing ovarian follicles as endpoints of safety tests, and the number of these structures is also of interest for other aspects of reproductive biology. Performing the counts microscopically on representative hematoxyli n & eosin-stained sections of ovary is tedious and error-prone. The ability to mark oocyte nuclei distinctly with an antibody significantly increases speed and accuracy of counting. We have identified a rabbit polyclonal antibody directed against a synthetic 14-amino acid sequence from human cytochrome P450 1B1 (CYP1B1) that unequivocally marks oocyte nuclei in addition to nuclei of some ovarian granulosa and theca cells. Follicles of all degrees of maturity are easily distinguished from ovarian background; ability to detect and identify primordial follicles is particularly enhanced. Excellent labeling was achieved with routine immunohistochemical procedures using an avidin-biotin-peroxidas e method on rat and mouse tissues fi xed in 10% neutral-buffered formalin. The antigenic peptide and the antiserum were produced by Alpha Diagnostics Incorporated, San Antonio, TX 78238 (www.4adi.com).


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