title: An immunohistochemical label to facilitate counting of ovarian follicles.
abstract: 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 identified 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.
authors.affiliation: Pathology Associates--a Charles River Company, National Center for Toxicological Research, Jefferson, Arkansas 72079, USA. lmuskhelishvili@nctr.fda.gov
authors.forename: Levan
authors.initials: L
authors.lastname: Muskhelishvili
authors.forename: Lisa D
authors.initials: LD
authors.lastname: Freeman
authors.forename: John R
authors.initials: JR
authors.lastname: Latendresse
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Bucci
mesh_terms: D000818:Animals; D000906:Antibodies; D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism*; D002452:Cell Count; D065633:Cytochrome P-450 CYP1B1; D005260:Female; D006107:Granulosa Cells / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D051379:Mice; D008810:Mice, Inbred C57BL; D009865:Oocytes / Q000378:metabolism; D006080:Ovarian Follicle / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013194:Staining and Labeling; D013799:Theca Cells / Q000378:metabolism
chemical_list: D000906:Antibodies; D001189:Aryl Hydrocarbon Hydroxylases; C585496:CYP1B1 protein, human; C585498:Cyp1b1 protein, mouse; C585499:Cyp1b1 protein, rat; D065633:Cytochrome P-450 CYP1B1