Regalado F, Esponda P, Nieto A
Centro de Biología Molecular (CSIC-UAM), Universidad Autónoma, Madrid, Spain.
Mol Reprod Dev. 1993 Dec;36(4):448-53. doi: 10.1002/mrd.1080360407.
We have compared the biosynthesis of secretory proteins in rabbit cauda epididymidis maintained for 15 days at abdominal temperature with that of the scrotal cauda. Explants from both situations were incubated in vitro in the presence of [35S] methionine, and the labelled proteins released into the incubation medium were analyzed by polyacrylamide gel electrophoresis. Body temperature specifically inhibited the synthesis of at least two polypeptides of 43 kDa and 21 kDa (designated EP21), whereas the synthesis of polypeptides of 80, 39, 31, and 24 kDa was increased. These changes resembled those produced by castration, but androgen treatment was not able to reverse the effect of body temperature. To confirm these observations, poly(A)+ RNA from the scrotal and the abdominal cauda respectively, was translated in vitro and the synthesized products were immunoprecipitated with an antibody against EP21 polypeptide. Both castration and body temperature strongly decreased the concentration of EP21 mRNA. In vivo testosterone administration restored the content of EP21 mRNA in cauda from castrated animals, but not in cauda maintained at body temperature. The changes observed might be related to the adverse effect of body temperature on sperm storage in the cauda epididymidis.
我们比较了在腹部温度下维持15天的兔附睾尾与阴囊附睾尾中分泌蛋白的生物合成情况。将两种情况下的外植体在[35S]甲硫氨酸存在下进行体外培养,并用聚丙烯酰胺凝胶电泳分析释放到培养液中的标记蛋白。体温特异性地抑制了至少两种43 kDa和21 kDa多肽(命名为EP21)的合成,而80、39、31和24 kDa多肽的合成增加。这些变化类似于去势所产生的变化,但雄激素处理无法逆转体温的影响。为了证实这些观察结果,分别从阴囊附睾尾和腹部附睾尾提取的聚腺苷酸加尾RNA(poly(A)+ RNA)进行体外翻译,合成产物用抗EP21多肽抗体进行免疫沉淀。去势和体温均强烈降低了EP21 mRNA的浓度。体内给予睾酮可恢复去势动物附睾尾中EP21 mRNA的含量,但不能恢复维持在体温下的附睾尾中的含量。观察到的这些变化可能与体温对附睾尾精子储存的不利影响有关。