Cameo M S, Blaquier J A
J Endocrinol. 1976 Apr;69(1):47-55. doi: 10.1677/joe.0.0690047.
The pattern of proteins in the soluble fraction of the cytoplasm of the rat epididymis was studied by acrylamide gel electrophoresis. The components of five distinct bands, labelled A, B, C, D and E, were found to be sensitive to changes in androgen in the blood. Castration for 14 days produced a sharp decrease in the colour intensity of bands B-E when stained with Amido black. After 21 days of castration, bands D and E were undetectable, bands B and C were severely diminished and band A was more intense. Seven days of replacement with testosterone (1 mg/day) induced a return towards a normal pattern. The degree of restoration was inversely proportional to the duration of castration. Quantitation by densitometry showed that the relative contributions of bands B-E to the region A-E were 61% in the control rat, only 27% after 21 days of castration and 35% when testosterone was given between days 14 and 21 of castration. The components of bands A-E are presumed to be proteins since the electrophoretic pattern was altered by digestion with pronase but not by ribonuclease, phospholipase C or neuraminidase. Epididymides from castrated and androgen-treated castrated rats were incubated with 14C- and 3H-labelled mixed amino acids respectively. After co-electrophoresis the ratio 3H: 14C rose from a baseline of 2-5 in band B, 32 in band C and 7 in bands D and E. Molecular weights were estimated as 27900 for B, 23100 for C and 34400 for D. Band A had the same electrophoretic mobility as serum albumin. Bands B and C were also present in testicular cytosol. Bands D and E were only found in the epididymis, localized mainly within the lumen of the tubules. Bands B-E increased with age during sexual maturation, bands D and E became detectable in the 20-day-old rats. Preliminary evidence indicates that the proteins in bands C, D and E can be removed from caput spermatozoa by washing.
通过丙烯酰胺凝胶电泳研究了大鼠附睾细胞质可溶性部分中的蛋白质模式。发现标记为A、B、C、D和E的五条不同条带的成分对血液中雄激素的变化敏感。去势14天后,用酰胺黑染色时,条带B - E的颜色强度急剧下降。去势21天后,条带D和E无法检测到,条带B和C严重减少,而条带A更明显。用睾酮(1毫克/天)替代7天导致恢复到正常模式。恢复程度与去势持续时间成反比。通过光密度测定法进行定量分析表明,条带B - E在区域A - E中的相对贡献在对照大鼠中为61%,去势21天后仅为27%,在去势第14天至21天给予睾酮时为35%。条带A - E的成分被推测为蛋白质,因为电泳模式在用链霉蛋白酶消化后发生改变,但用核糖核酸酶、磷脂酶C或神经氨酸酶处理则无变化。分别用14C和3H标记的混合氨基酸孵育去势大鼠和雄激素处理去势大鼠的附睾。共电泳后,条带B中3H:14C的比值从基线值2 - 5上升,条带C中为32,条带D和E中为7。估计条带B的分子量为27900,条带C为23100,条带D为34400。条带A的电泳迁移率与血清白蛋白相同。条带B和C也存在于睾丸细胞质溶胶中。条带D和E仅在附睾中发现,主要位于小管腔内。在性成熟过程中,条带B - E随年龄增加,在20日龄大鼠中可检测到条带D和E。初步证据表明,条带C、D和E中的蛋白质可通过洗涤从附睾头精子中去除。