Flow cytometric analysis of effects of 1,3-dinitrobenzene on rat spermatogenesis.
Exposure of 100-d old rats to 1,3-dinitrobenzene (m-DNB) at dosages up to 48 mg/kg resulted in disruption of spermatogenesis as measured by flow cytometry (FCM) of acridine orange-stained sperm and testis cells. One day (d 1) after a single exposure to 48 mg/kg m-DNB, FCM measurements of caput epididymal fluid cells demonstrated the presence of testicular germinal epithelial cells apparently sloughed off into the epididymis. Also, at d 1 after the same exposure, a decrease in pachytene spermatocytes was observed. By d 16 after exposure to 32 or 48 mg/kg, testicular damage was evidenced by an alteration of cell type ratios in FCM-analyzed populations of testicular cells. Extensive recovery of cell type ratios occurred by d 32. At d 16, dosages of 32 and 48 mg/kg caused alterations of sperm chromatin structure as determined by the flow cytometric sperm chromatin structure assay (SCSA); 48 mg/kg caused alterations at both d 16 and d 32. Exposure to m-DNB caused a dose response increase in percent sperm head morphology abnormalities (%ABN) assessed in cauda epididymal and vas sperm. A slightly higher correlation existed between dose and SCSA alpha t values (d 16, .78; p less than .01) than between dose and %ABN (d 16, .70; p less than .01). Also, a higher correlation existed between standard deviation of alpha t (SD alpha t) values and %ABN (.97; p less than .01) than between dose and %ABN (.70; p less than .01). This study demonstrated rapid and unique FCM procedures originally derived for reproductive toxicology studies in mice to be equally useful for studies in rats.
Olson Biochemistry Laboratories, South Dakota State University, Brookings 57007.
D P
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Evenson
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Janca
L K
LK
Jost
R K
RK
Baer
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Karabinus
D000818:Animals; D001835:Body Weight / Q000187:drug effects; D002843:Chromatin / Q000235:genetics; D004136:Dinitrobenzenes / Q000633:toxicity*; D004171:Diploidy; D004822:Epididymis / Q000187:drug effects; D005434:Flow Cytometry; D006238:Haploidy; D008297:Male; D009578:Nitrobenzenes / Q000633:toxicity*; D009929:Organ Size / Q000187:drug effects; D051381:Rats; D011919:Rats, Inbred Strains; D012260:Ribonucleases; D013010:Sonication; D013087:Spermatids / Q000187:drug effects; D013091:Spermatogenesis / Q000187:drug effects*; D013737:Testis / Q000187:drug effects; D014649:Vas Deferens / Q000187:drug effects
D002843:Chromatin; D004136:Dinitrobenzenes; D009578:Nitrobenzenes; C017906:3-dinitrobenzene; D012260:Ribonucleases