Flow cytometric analysis of effects of 1,3‐dinitrobenzene on rat spermatogenesis

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Abstract
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 acri‐dine 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 epididy‐mis. Also, at d 1 after the same exposure, a decrease in pachytene spermatocytes was observed. By d 76 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 76 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 α, values (d 76, .78; p < .01) than between dose and %ABN (d 16, .70; p < .01). Also, a higher correlation existed between standard deviation of αt, (SDαt,) values and %ABN (.97; p < .01) than between dose and %ABN (.70; p < .01). This study demonstrated rapid and unique FCM procedures originally derived for sreproductive toxicology studies in mice to be equally useful for studies in rats.