Identification of type IV collagenase in rat testicular cell culture: influence of peritubular-Sertoli cell interactions.
Identification of type IV collagenase in rat testicular cell culture: influence of peritubular-Sertoli cell interactions.
复制标题
大鼠睾丸细胞培养物中 IV 型胶原酶的鉴定：管周支持细胞相互作用的影响。
DOI：
10.1095/biolreprod43.6.956
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发表时间：
1990
影响因子：
3.6
通讯作者：
Byers,SW
中科院分区：
文献类型：
--
作者：
Sang,QX;Stetler-Stevenson,WG;Liotta,LA;Byers,SW
在睾丸中，小管周细胞（间充质）和支持细胞（上皮）之间的相互作用，以及细胞外基质的蛋白水解重塑，可能在睾丸发育，形态发生和精子发生中发挥核心作用。在这项研究中，我们证明，72 kDa的金属蛋白酶存在于大鼠支持细胞和支持-肾小管周细胞共培养液被激活旁路-氨基苯汞醋酸（p-APMA），以较低的分子量的形式，这表明它很可能是一个潜在的胶原酶。使用针对IV型胶原酶的三个不同结构域的抗体的免疫印迹显示72-kDa蛋白酶和76-kDa蛋白酶是IV型前胶原酶。单独培养的支持细胞产生基础水平的IV型胶原酶，其可以免疫定位在培养细胞的细胞质中。单独培养的管周细胞产生的IV型胶原酶少得多。然而，Sertoli和管周细胞在共培养产生IV型前胶原酶，并在培养物中主要由管周细胞，IV型胶原酶的活化形式检测到的酶谱和免疫印迹。在过渡期（从细胞附着到汇合）生长的细胞比在汇合期分泌更多的金属蛋白酶。相反，纤溶酶原激活剂水平不受培养时间的影响。这些结果表明，培养的大鼠睾丸细胞产生并分泌IV型胶原酶，并且这种酶和其他金属蛋白酶的分泌和激活受间充质细胞与上皮细胞的比例和培养时间的调节。
In the testis, interactions between peritubular cells (mesenchyme) and Sertoli cells (epithelium), together with proteolytic remodeling of the extracellular matrix, may play a central role in testicular development, morphogenesis, and spermatogenesis. In this study we demonstrate that a metalloproteinase of 72 kDa present in rat Sertoli cell and Sertoli-peritubular cell coculture medium is activated byp-aminophenylmercuric acetate (p-APMA) to a lower molecular mass form, indicating that it is likely to be a latent collagenase. Immunoblots using antibodies to three different domains of type IV collagenase show that the 72-kDa protease and a 76-kDa protease are type IV pro-collagenases. Sertoli cells cultured alone produce basal levels of type IV collagenase that can be immunolocalized in the cytoplasm of cultured cells. Peritubular cells cultured alone produce much less type IV collagenase. However, Sertoli and peritubular cells in coculture do produce type IV pro-collagenase, and in cultures consisting predominantly of peritubular cells, the activated form of type IV collagenase was detected by both zymography and immunoblotting. Cells growing during the transitional phase (from cell attachment to confluence) secrete more metalloproteinases than during the confluent phase. In contrast, plasminogen activator levels are unaffected by time in culture. These results show that rat testicular cells in culture produce and secrete type IV collagenase, and that the secretion and activation of this enzyme and other metalloproteases is regulated by the ratio of mesenchymal cells to epithelial cells and time in culture.