Bakalska Mariana, Atanassova Nina, Koeva Yvetta, Nikolov Boycho, Davidoff Michail
Institute of Experimental Morphology & Anthropology, Bulgarian Academy of Sciences, Sofia, Bulgaria.
Endocr Regul. 2004 Sep;38(3):103-10.
To carry out a detailed quantitative analysis of male germ cell apoptosis in seminiferous epithelium in a long period after EDS administration.
The apoptosis in adult rat testes was induced by a single i.p. injection of ethane dimethanesulphonate (EDS) in a dose of 75 mg/kg body weight. The TUNEL assay for in situ detection of apoptosis and quantitation of apoptotic germ were performed in testicular sections days 1, 3, 7, 14, 21 and 35 after EDS treatment. Plasma levels of testosterone (T) and luteinizing hormone (LH) were measured by RIA.
First signs of seminiferous epithelium regression were manifested by a marked increase in the number of apoptotic cells on 3rd day after EDS treatment. The maximal value of germ cell apoptosis was established on 7th day post EDS that coincided with lowest T levels. Later, until the end of investigated period, the elevated values of all investigated parameters for quantification of germ cell apoptosis decreased, but remained still higher as compared to control and, in addition, also T concentrations returned to normal range and their mean values were lower than these in controls. The pachytene spermatocytes and spermatids were the predominant cell types that underwent apoptosis after EDS treatment.
Quantitative patterns of germ cell death after testosterone deprivation reveal in advance the kinetic of germ cell depletion and regeneration in a long period after EDS. These new findings bring additional support to the concept that germ cell apoptosis is a hormonally regulated process. Induction of germ cell apoptosis by EDS could be considered as a result of differential alterations occurring in the main testicular cell types, more than one pathway being probably involved in that physiological cell death in the testis.
对给予EDS后长期生精上皮中雄性生殖细胞凋亡进行详细的定量分析。
通过腹腔注射75mg/kg体重的乙烷二甲磺酸盐（EDS）诱导成年大鼠睾丸凋亡。在EDS处理后的第1、3、7、14、21和35天，对睾丸切片进行原位检测凋亡和定量凋亡生殖细胞的TUNEL检测。通过放射免疫分析法测定血浆睾酮（T）和黄体生成素（LH）水平。
生精上皮退化的最初迹象表现为EDS处理后第3天凋亡细胞数量显著增加。生殖细胞凋亡的最大值在EDS处理后第7天确定，这与最低的T水平一致。之后，直到研究期结束，所有用于定量生殖细胞凋亡的研究参数的升高值均下降，但仍高于对照组，此外，T浓度也恢复到正常范围，但其平均值低于对照组。粗线期精母细胞和精子细胞是EDS处理后发生凋亡的主要细胞类型。
睾酮剥夺后生殖细胞死亡的定量模式预先揭示了EDS后长期生殖细胞耗竭和再生的动力学。这些新发现为生殖细胞凋亡是一个激素调节过程的概念提供了额外支持。EDS诱导生殖细胞凋亡可能是主要睾丸细胞类型发生差异改变的结果，睾丸中这种生理性细胞死亡可能涉及不止一条途径。
