The mouse testis is the source of various serine proteases and serine proteinase inhibitors (SERPINs): Serine proteases and SERPINs identified in Leydig cells are under gonadotropin regulation
The mouse testis is the source of various serine proteases and serine proteinase inhibitors (SERPINs): Serine proteases and SERPINs identified in Leydig cells are under gonadotropin regulation
复制标题
DOI：
10.1210/en.2006-0484
复制
发表时间：
2006-09-01
期刊：
影响因子：
4.8
通讯作者：
Le Magueresse-Battistoni, Brigitte
中科院分区：
文献类型：
--
作者：
Odet, Fanny;Verot, Adelie;Le Magueresse-Battistoni, Brigitte
通过RT-PCR研究了1、3和8周龄小鼠全睾丸中粗品和富集的生殖细胞组分、小鼠Leydig肿瘤细胞（mLTC-1）以及3和8周龄Leydig细胞和3周龄Sertoli细胞富集组分的原代培养物中各种丝氨酸蛋白酶和丝氨酸蛋白酶抑制剂（SERPIN）的发生。在睾丸蛋白酶谱中鉴定了新成员。在Leydig库中，发现了纤溶酶原激活剂尿激酶纤溶酶原激活剂（uPA）和组织纤溶酶原激活剂（8周龄细胞）、间质蛋白酶-2（mLTC-1）、激肽释放酶-21、SERPINA 5、SERPINB 2（原代培养物）和丝氨酸肽酶抑制剂Kunitz 2型（SPINT 2）的PCR产物。以类固醇生成急性调节蛋白（星星）为阳性对照，采用半定量RT-PCR方法研究促性腺激素的调节作用。在mLTC-1细胞中，人绒毛膜促性腺激素（hCG）通过cAMP下调间质蛋白酶-2、激肽释放酶-21、SPINT 2和SERPINA 5，而上调uPA及其受体。在hCG暴露1-8小时后观察到短暂的阳性效应，6小时后首次出现的阴性效应持续48小时。在原代培养物中证实了hCG诱导的作用。此外，SERPINB 2在原代培养物中被hCG增强。添加胰蛋白酶或蛋白酶抑制剂没有改变hCG诱导的星星激增。由于hCG调节蛋白酶和SERPIN（而睾酮不调节），因此它可以改变Leydig细胞的蛋白水解平衡，从而改变细胞外基质成分的代谢。因此，即使早期hCG诱导的uPA和星星激增之间的直接相互作用是不可能的，我们的数据与文献一起表明，细胞外基质蛋白改变Leydig细胞类固醇生成。
The occurrence of various serine proteinases and serine proteinases inhibitors (SERPINs) was investigated by RT-PCR in whole testes of 1-, 3-, and 8-wk-old mice in crude and enriched germ cell fractions, mouse Leydig tumor cells (mLTC-1), and primary cultures of 3- and 8-wk-old enriched fractions of Leydig cells and 3-wk-old Sertoli cells. New members were identified in the testis protease repertoire. Within the Leydig repertoire, a PCR product was found for plasminogen activators urokinase plasminogen activator (uPA) and tissue plasminogen activator (8-wk-old cells), matriptase-2 (mLTC-1), kallikrein-21, SERPINA5, SERPINB2 (primary cultures), and serine peptidase inhibitor Kunitz type 2 (SPINT2). The gonadotropin regulation was explored by semiquantitative RT-PCR, using steroidogenic acute regulatory protein (StAR) as a positive control. Matriptase-2, kallikrein-21, SPINT2, and SERPINA5 were down-regulated, whereas uPA and its receptor were up-regulated by human chorionic gonadotropin (hCG) via cAMP in the mLTC-1 cells. Positive effects were observed transiently after 1-8 h of hCG exposure, and negative effects, first evidenced after 6 h, lasted 48 h. The hCG-induced effects were confirmed in primary cultures. In addition, SERPINB2 was augmented by hCG in primary cultures. Addition of either trypsin or protease inhibitors did not alter the hCG-induced surge of StAR. Because hCG regulated proteases and SERPINs (whereas testosterone did not), it could alter the proteolytic balance of Leydig cells and consequently the metabolism of extracellular matrix components. Therefore, even though a direct interplay between the early hCG-induced surge of uPA and StAR is unlikely, our data together with the literature suggest that extracellular matrix proteins alter Leydig cell steroidogenesis.