Frederik P M, Klepper D, Van der Vusse G J, Van der Molen H J
Mol Cell Endocrinol. 1976 Jun-Jul;5(1-2):123-36. doi: 10.1016/0303-7207(76)90075-7.
Localization of radioactive steroids in rat testis was studied by autoradiography of tissue section. For autoradiography small tissue samples were frozen, freeze-dried under vacuum, fixed with osmium vapor and embedded in epon. The transfer of radioactive steroids was studied after in vitro perfusion of radioactive steroids in the testes isolated from hypophysectomized animals, thereby excluding the interference of endogenous steroids. Quantitative autoradiography on the basis of grain densities after perfusion of testes with tritiated pregnenolone or testosterone, revealed an accumulation of the label in the Leydig cell cytoplasm. After longer perfusion periods the amount of label in the seminiferous tubules increased and a preferential localization was observed in the basal cytoplasm of Sertoli cells and in lipid droplets. Perfusion of testes with estradiol-17 beta resulted in a distinctly different pattern of radioactivity in the autoradiographs. A high labeling of the Leydig cell nuclei was observed in combination with a low general labeling of all the other cell structures. The results suggest that different steroids are localized in different specific areas of the rat testes in vivo.
通过组织切片放射自显影术研究了放射性甾体在大鼠睾丸中的定位。对于放射自显影,将小组织样本冷冻,在真空下冷冻干燥,用锇蒸汽固定并包埋在环氧树脂中。在对从垂体切除动物分离的睾丸进行放射性甾体体外灌注后,研究了放射性甾体的转运,从而排除了内源性甾体的干扰。在用氚化孕烯醇酮或睾酮灌注睾丸后,基于颗粒密度的定量放射自显影显示,标记物在睾丸间质细胞胞质中积累。灌注时间延长后,生精小管中的标记物数量增加,并且在支持细胞的基底胞质和脂滴中观察到优先定位。用雌二醇-17β灌注睾丸导致放射自显影片中放射性分布明显不同。观察到睾丸间质细胞核高度标记,而所有其他细胞结构的总体标记较低。结果表明,不同的甾体在大鼠睾丸的不同特定区域中定位。