ToxSci Advance Access originally published online on October 25, 2006
Toxicological Sciences 2007 95(1):249-256; doi:10.1093/toxsci/kfl141
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Stage-Related Increase in the Proportion of Apoptotic Germ Cells and Altered Frequencies of Stages in the Spermatogenic Cycle Following Gestational, Lactational, and Direct Exposure of Male Rats to p-Nonylphenol
* Department of Physiology, Faculty of Health Sciences, University of Pretoria, P.O. Box 2034, Pretoria 0001, SOUTH AFRICA
School of Health Systems and Public Health, Faculty of Health Sciences, University of Pretoria, P.O. Box 2034, Pretoria 0001, SOUTH AFRICA
Department of Urology, Faculty of Health Sciences, University of Pretoria, Pretoria 0001, South Africa
1 To whom correspondence should be addressed at Department of Physiology, Faculty of Health Sciences, University of Pretoria, P.O. Box 2034 Pretoria 0001, South Africa. Fax: +27 (0) 321-1679. E-mail: leon.mcclusky{at}med.up.ac.za.
Received August 2, 2006; accepted October 23, 2006
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Abstract |
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The cumulative effects of environmental toxicants, for example, the alkylphenol, para-nonylphenol (p-NP) are of concern. Our previous study showed that p-NP reduced several testicular morphometric parameters, including sperm counts. The present study reexamined material collected in that study to determine the mechanistic basis of p-NP action on spermatogenic development in the offspring. Seven-day pregnant Sprague-Dawley rats were treated with vehicle or 100 or 250 mg/kg p-NP through gestation, lactation and afterward directly to all male offspring until 10 weeks of age. Both doses of p-NP significantly (P < 0.02) increased the number of germ cells with in situ end-labeled fragmented DNA (TUNEL positive) by 1.9-fold and 1.7-fold, respectively, and specifically in stages XII–XIV and I–III. TUNEL-labeling was, however, selective, and excluded labeling of basal cells with apoptotic morphology. Cleaved caspase-3 immunohistochemistry strongly labeled basal cells (spermatogonia and early spermatocytes) with condensed marginated chromatin but not degenerate germ cells lacking definitive nuclear material found throughout the epithelium. Only the caspase index (ratio of number of caspase positive to number of degenerate cells) of the 100-mg/kg p-NP group was significantly (p < 0.05) threefold greater than controls. Whereas both doses and either 250 or 100 mg/kg treatment alone significantly (p < 0.002) reduced the frequencies (duration) of stages I–III, VII–VIII, and late VIII–IX (spermiating and recently spermiated tubules), respectively, both doses significantly (p < 0.002) increased the frequencies of stages IV–VI and all stages containing late-stage spermatocytes (XII–XIII) and meiotic cell divisions (XIV). Thus, p-NP, an environmentally persistent xenoestrogen, insidiously alters the spermatogenic cycle and spermatogenic process in male offspring.
Key Words: spermatogenesis; p-nonylphenol; cleaved caspase-3; stages; TUNEL.
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