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Toxicologic Pathology
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An Immunohistochemical Label to Facilitate Counting of Ovarian Follicles

Levan Muskhelishvili

Pathology Associates-A Charles River Company, National Center for Toxicological Research, Jefferson, Arkansas, USA, lmuskhelishvili{at}nctr.fda.gov.

Lisa D. Freeman

Pathology Associates-A Charles River Company, National Center for Toxicological Research, Jefferson, Arkansas, USA

John R. Latendresse

Pathology Associates-A Charles River Company, National Center for Toxicological Research, Jefferson, Arkansas, USA

Thomas J. Bucci

Pathology Associates-A Charles River Company, National Center for Toxicological Research, Jefferson, Arkansas, USA

U.S. and internationally harmonized Health Effects Test Guidelines for Reproduction and Fertility Effects 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 hematoxylin and eosin (H&E)-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 identifi ed a rabbit polyclonal antibody directed against a synthetic 14-amino acid sequence from human cytochrome P-450 1B1 (CYP1B1) that unequivocally marks rodent 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. High-contrast and high-resolution labeling was achieved with routine immunohistochemical procedures using an avidin—biotin —peroxidase method on rat and mouse tissues fixed in 10% neutral buffered formalin.

Key Words: CYP1B1 • cytochrome P-450 • immunohistochemistry • ovarian follicles • rodent.

Toxicologic Pathology, Vol. 30, No. 3, 400-402 (2002)
DOI: 10.1080/01926230252929981
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This article has been cited by other articles:


Home page
 Toxicol PatholHome page
C. A. Picut, C. L. Swanson, K. L. Scully, V. C. Roseman, R. F. Parker, and A. K. Remick
Ovarian Follicle Counts Using Proliferating Cell Nuclear Antigen (PCNA) and Semi-Automated Image Analysis in Rats
Toxicol Pathol, July 1, 2008; 36(5): 674 - 679.
[Abstract] [Full Text] [PDF]

Home page
 Toxicol PatholHome page
L. Muskhelishvili, S. K. Wingard, and J. R. Latendresse
Proliferating Cell Nuclear Antigen--A Marker for Ovarian Follicle Counts
Toxicol Pathol, April 1, 2005; 33(3): 365 - 368.
[Abstract] [Full Text] [PDF]

Home page
 Toxicol PatholHome page
STP Ovary Evaluation Working Group, K. S. Regan, J. M. Cline, D. Creasy, B. Davis, G. L. Foley, L. Lanning, J. R. Latendresse, S. Makris, D. Morton, et al.
STP Position Paper: Ovarian Follicular Counting in the Assessment of Rodent Reproductive Toxicity
Toxicol Pathol, April 1, 2005; 33(3): 409 - 412.
[Full Text] [PDF]