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Title: Evaluation of ovotoxicity induced by 7, 12-dimethylbenz[a]anthracene and its 3,4-diol metabolite utilizing a rat in vitro ovarian culture system

Abstract

The polycyclic aromatic hydrocarbon 7, 12-dimethylbenz[a]anthracene, (DMBA), targets and destroys all follicle types in rat and mouse ovaries. DMBA requires bioactivation to DMBA-3,4-diol-1,2-epoxide for ovotoxicity via formation of the intermediate, DMBA-3,4-diol (catalyzed by microsomal epoxide hydrolase; mEH). mEH was shown to be involved in DMBA bioactivation for ovotoxicity induction in B6C3F{sub 1} mouse ovaries. The current study compared DMBA and DMBA-3,4-diol mediated ovotoxicity, and investigated mEH involvement in DMBA-3,4-diol bioactivation in Fischer 344 (F344) rat ovary. F344 postnatal day (PND) 4 rat ovaries were cultured in vehicle control or media containing 1) DMBA or DMBA-3,4-diol (12.5 nM - 1 {mu}M; 15 days); 2) DMBA (1 {mu}M; 6 h - 15 days); and 3) DMBA (1 {mu}M) or DMBA-3,4-diol (75 nM) {+-} the mEH activity inhibitor cyclohexene oxide (CHO; 2 mM; 4 days). Ovaries were histologically evaluated and mEH mRNA and protein were measured by reverse transcriptase PCR or Western blotting, respectively. Ovotoxicity following 15 days of culture occurred (P < 0.05) at lower concentrations of DMBA-3,4-diol (12.5 nM - primordial; 75 nM - primary) than DMBA (75 nM - primordial; 375 nM - primary). The temporal pattern of mEH expression following DMBA exposure showed mRNA up-regulation (P < 0.05) onmore » day 2, with increased protein (P < 0.05) on day 4, the earliest time of observed follicle loss (P < 0.05). mEH inhibition prevented DMBA-induced, but not DMBA-3,4-diol-induced ovotoxicity. These results demonstrate a conserved response in mice and rats for ovarian mEH involvement in DMBA bioactivation to its ovotoxic, 3,4-diol-1,2-epoxide form.« less

Authors:
 [1];  [1];  [2];  [1]
  1. Department of Physiology, University of Arizona, Tucson, AZ 85745 (United States)
  2. Department of Pharmacology, University of Arizona, Tucson, AZ 85745 (United States)
Publication Date:
OSTI Identifier:
21182722
Resource Type:
Journal Article
Journal Name:
Toxicology and Applied Pharmacology
Additional Journal Information:
Journal Volume: 234; Journal Issue: 3; Other Information: DOI: 10.1016/j.taap.2008.10.009; PII: S0041-008X(08)00439-0; Copyright (c) 2008 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0041-008X
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ANTHRACENE; DIMETHYLBENZANTHRACENE; EPOXIDES; EVALUATION; GLYCOLS; IN VITRO; MICE; OVARIES; OXIDES; POLYCYCLIC AROMATIC HYDROCARBONS; POLYMERASE CHAIN REACTION; PROTEINS; RATS

Citation Formats

Igawa, Yoshiyuki, BioPharma Center, Asubio Pharma Co., Ltd., Gunma, Keating, Aileen F., Rajapaksa, Kathila S, Amgen Inc., Toxicology Department, Thousand Oaks, CA, 91320-1799, Sipes, I Glenn, and Hoyer, Patricia B. Evaluation of ovotoxicity induced by 7, 12-dimethylbenz[a]anthracene and its 3,4-diol metabolite utilizing a rat in vitro ovarian culture system. United States: N. p., 2009. Web. doi:10.1016/j.taap.2008.10.009.
Igawa, Yoshiyuki, BioPharma Center, Asubio Pharma Co., Ltd., Gunma, Keating, Aileen F., Rajapaksa, Kathila S, Amgen Inc., Toxicology Department, Thousand Oaks, CA, 91320-1799, Sipes, I Glenn, & Hoyer, Patricia B. Evaluation of ovotoxicity induced by 7, 12-dimethylbenz[a]anthracene and its 3,4-diol metabolite utilizing a rat in vitro ovarian culture system. United States. https://doi.org/10.1016/j.taap.2008.10.009
Igawa, Yoshiyuki, BioPharma Center, Asubio Pharma Co., Ltd., Gunma, Keating, Aileen F., Rajapaksa, Kathila S, Amgen Inc., Toxicology Department, Thousand Oaks, CA, 91320-1799, Sipes, I Glenn, and Hoyer, Patricia B. 2009. "Evaluation of ovotoxicity induced by 7, 12-dimethylbenz[a]anthracene and its 3,4-diol metabolite utilizing a rat in vitro ovarian culture system". United States. https://doi.org/10.1016/j.taap.2008.10.009.
@article{osti_21182722,
title = {Evaluation of ovotoxicity induced by 7, 12-dimethylbenz[a]anthracene and its 3,4-diol metabolite utilizing a rat in vitro ovarian culture system},
author = {Igawa, Yoshiyuki and BioPharma Center, Asubio Pharma Co., Ltd., Gunma and Keating, Aileen F. and Rajapaksa, Kathila S and Amgen Inc., Toxicology Department, Thousand Oaks, CA, 91320-1799 and Sipes, I Glenn and Hoyer, Patricia B},
abstractNote = {The polycyclic aromatic hydrocarbon 7, 12-dimethylbenz[a]anthracene, (DMBA), targets and destroys all follicle types in rat and mouse ovaries. DMBA requires bioactivation to DMBA-3,4-diol-1,2-epoxide for ovotoxicity via formation of the intermediate, DMBA-3,4-diol (catalyzed by microsomal epoxide hydrolase; mEH). mEH was shown to be involved in DMBA bioactivation for ovotoxicity induction in B6C3F{sub 1} mouse ovaries. The current study compared DMBA and DMBA-3,4-diol mediated ovotoxicity, and investigated mEH involvement in DMBA-3,4-diol bioactivation in Fischer 344 (F344) rat ovary. F344 postnatal day (PND) 4 rat ovaries were cultured in vehicle control or media containing 1) DMBA or DMBA-3,4-diol (12.5 nM - 1 {mu}M; 15 days); 2) DMBA (1 {mu}M; 6 h - 15 days); and 3) DMBA (1 {mu}M) or DMBA-3,4-diol (75 nM) {+-} the mEH activity inhibitor cyclohexene oxide (CHO; 2 mM; 4 days). Ovaries were histologically evaluated and mEH mRNA and protein were measured by reverse transcriptase PCR or Western blotting, respectively. Ovotoxicity following 15 days of culture occurred (P < 0.05) at lower concentrations of DMBA-3,4-diol (12.5 nM - primordial; 75 nM - primary) than DMBA (75 nM - primordial; 375 nM - primary). The temporal pattern of mEH expression following DMBA exposure showed mRNA up-regulation (P < 0.05) on day 2, with increased protein (P < 0.05) on day 4, the earliest time of observed follicle loss (P < 0.05). mEH inhibition prevented DMBA-induced, but not DMBA-3,4-diol-induced ovotoxicity. These results demonstrate a conserved response in mice and rats for ovarian mEH involvement in DMBA bioactivation to its ovotoxic, 3,4-diol-1,2-epoxide form.},
doi = {10.1016/j.taap.2008.10.009},
url = {https://www.osti.gov/biblio/21182722}, journal = {Toxicology and Applied Pharmacology},
issn = {0041-008X},
number = 3,
volume = 234,
place = {United States},
year = {2009},
month = {2}
}