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

• Published in: Journal of toxicology and environmental health Vol. 28; no. 1; p. 81
• Main Authors: Evenson, D P, Janca, F C, Jost, L K, Baer, R K, Karabinus, D S
• Format: Journal Article
• Language: English
• Published: United States 01.09.1989
• Subjects: Animals; Body Weight - drug effects; Chromatin - genetics; Dinitrobenzenes - toxicity; Diploidy; Epididymis - drug effects; Flow Cytometry; Haploidy; Male; Nitrobenzenes - toxicity; Organ Size - drug effects; Rats; Rats, Inbred Strains; Ribonucleases; Sonication; Spermatids - drug effects; Spermatogenesis - drug effects; Testis - drug effects; Vas Deferens - drug effects
• ISSN: 0098-4108
• DOI: 10.1080/15287398909531330

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.