This Article Abstract Free Full Text (Free PDF) Free Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Muskhelishvili, L. Articles by Bucci, T. J. Search for Related Content PubMed PubMed Citation Articles by Muskhelishvili, L. Articles by Bucci, T. J. Social Bookmarking                         What's this?

An Immunohistochemical Label to Facilitate Counting of Ovarian Follicles

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.

References

• Bolon B., Bucci TJ, Warbritton AR, Chen JJ, Mattison DR, Heindel JJ (1997). Differential follicle counts as a screen for chemically induced ovarian toxicity in mice: Results from continuous breeding bioassays. Fundam Appl Toxicol 39: 1—10.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Bucci TJ, Bolon B., Warbritton AR, Chen JJ, Heindel JJ (1997). Infl uence of sampling on the reproducibility of ovarian follicle counts in mouse toxicity studies. Reprod Toxicol 11: 689—696.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Crespi CL, Penman BW, Steimel DT, Smith T., Yang CS, Sutter TR (1997). Development of a human lymphoblastoid cell line constitutively expressing human CYP1B1 cDNA: Substrate specifi city with model substrates and promutagens. Mutagenesis 12: 83—89.[Web of Science][Medline] [Order article via Infotrieve]
• Gonzales F. (1988). The molecular biology of cytochrome P450s. Pharmacol Rev 40: 243—288.[Web of Science][Medline] [Order article via Infotrieve]
• Guengerich FP (1994). Analysis and characterization of enzymes. In: Principles and methods of toxicology, Hayes AW (ed). Raven Press, New York, pp 1259—1313.
• Guengerich FP, Shimada T. (1991). Oxidation of toxic and carcinogenic chemicals by human cytochrome P-450 enzymes. Chem Res Toxicol 4: 391—407.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Hakkola J., Pasanen M.,Pelkonen O., Hukkanen J., Evisalmi S., Antilla S., Rane A., Mantyla M., Purkunen R., Saarikoski S., Tooming M., Raunio H. (1997). Expression of CYP1B1 in human adult and fetal tissues and differential inducibility of CYP1B1 and CYP1A1 by Ah receptor ligands in human placenta and cultured cells. Carcinogenesis 18: 391—397.[Abstract/Free Full Text]
• Hayes CL, Spink DC, Spink BC, Cao JQ, Walker NJ, Sutter TR (1996). 17β-Estradiol hydroxylation catalyzed by human cytrochrome P450 1B1. Proc Nat Acad Sci USA 93: 9776—9781.[Abstract/Free Full Text]
• Huang ZQ, Fasco MJ, Figge HL, Keyomarsi K., Kaminsky LS (1996). Expression of cytochrome P450 in human breast tissue and tumors. Drug Metab Dispos 24: 899—905.[Abstract]
• Liehr JG, Ricci MJ, Jefcoate CR, Hannigan EV, Hokanson JA, Zhu BT (1995). 4-Hydroxylation of estradiol by human uterine myometrium and myoma microsomes: Implication for the uterine tumorigenesis. Proc Nat Acad Sci USA 92: 9220—9224.[Abstract/Free Full Text]
• Marcinkiewicz JI, Krishna A., Cheung CMY, Terranova PF (1994). Oocyte tumor necrosis factor alpha: Localization in the neonatal ovary and throughout follicular development in the adult rat. Biol Reprod 50: 1251— 1260.
• Mattison DR,Thomford PJ (1989). The mechanismof action of reproductive toxicants. Toxicol Pathol 17: 364—376.[Web of Science][Medline] [Order article via Infotrieve]
• Muskhelishvili L., Thompson PA, Kusewitt DF, Wang C., Kadlubar FF (2000). In situ hybridization and immunohistochemical analysis of cytochrome P450 1B1 expression in human normal tissues. J Histochem Cytochem 49(2): 229—236.[Web of Science]
• Oktay K., Schenken RS, Nelson JF (1995). Proliferating cell nuclear antigen marks the initiation of follicular growth in the rat. Biol Reprod 53: 295—301.[Abstract]
• Park BK, Pirmohamed M., Kitteringham NR (1995). The role of cytochrome P450 enzymes in hepatic and extrahepatic human drug toxicity. Pharmacol Ther 68:385—424.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Rieder CRM, Ramsden DB, Williams AC (1998). Cytochrome P450 1B1 mRNA in the human central nervous system. J Clin Pathol Mol Pathol 51:138—142.[Abstract]
• Shimada T., Hayes CL, Yamazaki H., Shantu A., Hecht SS, Guengerich FP, Sutter TR (1996). Activation of chemically diverse procarcinogen s by human cytochrome P-450 1B1. Cancer Res 56: 2979—2984.[Abstract/Free Full Text]
• Skinner SM, Dunbar BS (1992). Localization of a carbohydrate antigen associated with growing oocytes and ovarian surface epithelium. J Histochem Cytochem 40: 1031—1036.[Abstract]
• Smith BJ, Plowchalk DR, Sipes IG, Mattison DR (1991). Comparison of random and serial sections in assessment of ovarian toxicity. Reprod Toxicol 5: 379—383.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Spink DC, Hayes CL, Young NR, Christou M., Sutter TR, Jefcoate CR, Gierthy JF (1994). The effects of 2,3,7,8-tetrachlorodibenzo -p-dioxin on estrogen metabolism in MCF-7 breast cancer cells: Evidence for induction of a novel 17β—estradiol 4-hydroxylase. J Steroid Biochem Mol Biol 51: 251—258.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Sutter TR, Tang YM, Hayes CL, Wo YY, Jabs EW, Li X., Yin H., Cody CW, Greenlee WF (1994). Complete cDNA sequence of a human dioxin-inducible mRNA identifi es a new gene subfamily of cytochrome p450 that maps to chromosome 2. J Bio Chem 269: 13092—13099.[Abstract/Free Full Text]
• Tang YM, Chen GF, Thompson PA, Lin D., Lang NP, Kadlubar FF (1999). Development of an antipeptide antibody that binds to the c-terminal region of human CYP1B1. Drug Metab Dispos 27: 274—280.[Abstract/Free Full Text]
• Vadlamuri SV, Glover DD, Turner T., Sarkar MA (1998). Regiospecific expression of cytochrome P4501A1 and 1B1 in human uterine tissue. Cancer Lett 122: 143—150.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]

Toxicologic Pathology, Vol. 30, No. 3, 400-402 (2002)
DOI: 10.1080/01926230252929981


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				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]

				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]

				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]

This Article Abstract Free Full Text (Free PDF) Free Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Muskhelishvili, L. Articles by Bucci, T. J. Search for Related Content PubMed PubMed Citation Articles by Muskhelishvili, L. Articles by Bucci, T. J. Social Bookmarking                         What's this?