Protein secretory patterns of rat Sertoli and peritubular cells are influenced by culture conditions.
Protein secretory patterns of rat Sertoli and peritubular cells are influenced by culture conditions.
复制标题
大鼠支持细胞和管周细胞的蛋白质分泌模式受培养条件的影响。
DOI：
10.1095/biolreprod35.1.239
复制
发表时间：
1986
影响因子：
3.6
通讯作者：
Tres,LL
中科院分区：
文献类型：
--
作者：
Kierszenbaum,AL;Crowell,JA;Shabanowitz,RB;DePhilip,RM;Tres,LL
采用双向凝胶电泳法和放射自显影技术相结合的方法，将支持细胞和肾小管周围细胞培养物中的分泌蛋白质与生精小管段培养上清液中观察到的蛋白质相关联。培养的支持细胞和生精小管中的支持细胞分泌三种蛋白质，分别命名为S70(Mr 72,000-70,000)、S45(Mr 45,000)和S35(Mr 35,000)。培养的支持细胞、肾小管周围细胞和培养的生精小管分泌两种蛋白质，分别命名为SP1(Mr 42,000)和SP2(Mr 50,000)。SP1和S45具有相似的MR，但等电点(PI)不同。培养的肾小管周围细胞分泌一种命名为P40(Mr 40,000)的蛋白质，这种蛋白质也在完整的生精小管中可见，但在没有肾小管周细胞壁的生精小管中则不存在。然而，大量的高MR蛋白仅在培养的小管周细胞的培养液中观察到，而在完整的生精小管的孵育液中没有观察到。培养条件影响培养的肾小管周细胞的形态和蛋白质分泌模式。当将肾小管周细胞置于无血清(有或无激素和生长因子)的培养液中时，表现为扁平-星形转变的细胞，在培养液中积累各种蛋白质，当这些细胞在添加血清的培养基中维持时，这些蛋白质不那么明显。在卵泡刺激素(FSH)刺激的生精小管段的孵育液中，也观察到了两种先前在培养的Sertoli细胞培养上清液中发现的FSH刺激的分泌蛋白(命名为SCm1和SCm2)。本研究结果表明，虽然培养的支持细胞和小管周细胞也能合成和分泌蛋白质，但在培养的生精小管节段中也观察到了蛋白质的合成和分泌，但花药蛋白质组缺乏生精小管的相关性。我们的观察应该有助于在培养中实现Sertoli和肾小管周围细胞的分化功能状态，以及选择分泌蛋白来评估它们在睾丸功能中可能的生物学作用。
An approach combining two-dimensional gel electrophoresis and autoradiography was used to correlate patterns of secretory proteins in cultures of Sertoli and peritubular cells with those observed in the incubation medium from segments of seminiferous tubules. Sertoli cells in culture and in seminiferous tubules secreted three proteins designated S70 (Mr72,000–70,000), S45 (Mr45,000), and S35 (Mr35,000). Cultured Sertoli and peritubular cells and incubated seminiferous tubules secreted two proteins designated SP1 (Mr42,000) and SP2 (Mr50,000). SP1 and S45 have similar Mrbut differ from each other in isoelectric point (pI). Cultured peritubular cells secreted a protein designated P40 (Mr40,000) that was also seen in intact seminiferous tubules but not in seminiferous tubules lacking the peritubular cell wall. However, a large number of high-Mrproteins were observed only in the medium of cultured peritubular cells but not in the incubation medium of intact seminiferous tubules. Culture conditions influence the morphology and patterns of protein secretion of cultured peritubular cells. Peritubular cells that display a flat-stellate shape transition when placed in culture medium free of serum (with or without hormones and growth factors), accumulate various proteins in the medium that are less apparent when these cells are maintained in medium supplemented with serum. Two secretory proteins stimulated by follicle-stimulating hormone (FSH) (designated SCm1 and SCm2) previously found in the medium of cultured Sertoli cells, were also observed in the incubation medium of seminiferous tubular segments stimulated by FSH. Results of this study show that, although cultured Sertoli and peritubular cells synthesize and secrete proteins also observed in segments of incubated seminiferous tubules anther group of proteins lacks seminiferous tubular correlates. Our observations should facilitate efforts to achieve a differentiated functional state of Sertoli and peritubular cells in culture as well as to select secretory proteins for assessing their possible biological role in testicular function.