Two-dimensional electrophoresis of proteins in principal cells, spermatozoa, and fluid associated with the rat epididymis.
Two-dimensional electrophoresis of proteins in principal cells, spermatozoa, and fluid associated with the rat epididymis.
复制标题
大鼠附睾主细胞、精子和液体中蛋白质的二维电泳。
作者：
R. Shabanowitz;G. Killian
通过二维电泳对附睾不同区域的精子、液体和主细胞进行了表征。结扎输精管36 h后收集睾丸网液，经输精管逆行灌注收集附睾尾液。精子从睾丸和附睾组织中分离出来后采集。附睾经酶解和离心洗脱后恢复主细胞。采用二维聚丙烯酰胺凝胶电泳制备各样品的蛋白质图谱。将睾丸网液中发现的蛋白质与附睾尾液中发现的蛋白质进行比较，发现其组成发生了巨大变化，包括特定蛋白质的丢失、增加或保留，以及某些蛋白质相对浓度的变化。在16、23和34 kDa处检测到突出的附睾尾液蛋白，可能是由附睾上皮贡献的。附睾过境后，水溶性精子蛋白的数量明显减少。附睾头和附睾体的附睾上皮主细胞的蛋白质组成也存在差异，这表明这些附睾区域存在功能差异。特别令人感兴趣的是，在所有精子样本、液体以及头和体主细胞中都发现了一种大约20-23 kDa的显著蛋白质。然而，在输精管结扎36小时后，附睾尾部精子中不存在该蛋白。出输精管结扎后精子中这种蛋白质的迅速丢失表明，这种手术干预可能会影响居住在附睾内的精子。
Spermatozoa, fluids, and principal cells from different regions of the epididymis were characterized by two-dimensional electrophoresis. Rete testis fluid was collected after 36-h efferent duct ligation, and cauda epididymal fluid was collected by retrograde perfusion through the vas deferens. Spermatozoa were collected after their exudation from minced caput and corpus epididymal tissue. Principal cells were recovered after enzymatic disaggregation and centrifugal elutriation of epididymides. Two-dimensional polyacrylamide gel electrophoresis was used to prepare protein profiles of all samples. Comparison of the proteins found in rete testis fluid versus those found in cauda epididymal fluid revealed a dramatic change in composition, including the loss, addition, or retention of specific proteins as well as changes in the relative concentrations of certain proteins. Prominent cauda epididymal fluid proteins, possibly contributed by the epididymal epithelium, were detected at 16, 23, and 34 kDa. After epididymal transit, a considerable decrease was observed in the number of aqueous-soluble sperm proteins. Differences in the protein composition of epididymal epithelial principal cells from the caput versus corpus epididymidis were also noted, suggesting that functional differences exist for these epididymal regions. Of particular interest was the occurrence of a prominent protein of approximately 20-23 kDa found in all sperm samples, in fluids, and in caput and corpus principal cells. However, this protein was absent in cauda epididymal sperm after 36-h efferent duct ligation. The rapid loss of this protein from sperm after efferent duct ligation suggests that this surgical intervention may affect spermatozoa residing within the epididymis.
灌注头附睾小管蛋白质的合成和分泌，以及分泌蛋白质与精子的关联。
影响因子：
3.6
作者：
Klinefelter,GR;Hamilton,DW
通讯作者：
Hamilton,DW