Spruill W A, Steiner A L, Earp H S
J Clin Invest. 1978 Sep;62(3):568-76. doi: 10.1172/JCI109162.
The relationship between the subcellular distribution of guanylate cyclase and tissue guanosine-3',5'-cyclic monophosphate (cGMP) levels was investigated in rat testes after surgically induced unilateral cryptorchidism. Placement of one of a testis pair in the abdominal cavity results in loss of testicular weight and function in the abdominal testis whereas the remaining scrotal testis appears to be functionally normal. Within 5 days after surgery, tissue cGMP levels were increased by twofold in the abdominal testis. A fourfold elevation was noted from 10 to 30 days after surgery. Whereas the homogenate guanylate cyclase activity was only slightly elevated 10 and 20 days postoperatively, a 200% increase in the soluble guanylate cyclase activity was seen at 5 days. Between 10 and 30 days, the rise in activity was >250% (P < 0.01). An increase in soluble guanylate cyclase activity was noted when the data were expressed as per milligram protein, per milligram DNA or per whole testis. Conversely, particulate guanylate cyclase activity was reduced by 40% in the cryptorchid testis. Kinetic analysis of the soluble enzyme prepared from abdominal and scrotal testes yielded linear Line-weaver-Burke plots for both enzyme preparations with an identical K(m) for guanosine triphosphate, but a three-fold higher maximal velocity for the abdominal enzyme. When the soluble guanylate cyclases from both testes were mixed and assayed together, the activities were additive rather than exhibiting synergism or inhibition. These experiments indicate that the altered V(max) is not due to a transferable activator or inhibitor.An immunocytochemical technique was used to assess the cell type in which the alterations in cGMP metabolism occurred. Comparison of the scrotal and abdominal testes revealed that the abdominal testis exhibited enhanced cGMP immunofluorescence within the cells lining the inner aspect of the seminiferous tubule as well as tubular elements and interstitial cells. Thus, it is inferred that the correlated changes in soluble guanylate cyclase activity and cGMP levels occur in several of the cell types that remain viable within the cryptorchid testis.
在手术诱导单侧隐睾后的大鼠睾丸中,研究了鸟苷酸环化酶的亚细胞分布与组织鸟苷 - 3',5'-环磷酸(cGMP)水平之间的关系。将一对睾丸中的一个置于腹腔会导致腹腔内睾丸的重量和功能丧失,而其余的阴囊内睾丸似乎功能正常。手术后5天内,腹腔内睾丸的组织cGMP水平增加了两倍。术后10至30天观察到增加了四倍。虽然匀浆鸟苷酸环化酶活性在术后10天和20天仅略有升高,但在5天时可溶性鸟苷酸环化酶活性增加了200%。在10至30天之间,活性升高>250%(P < 0.01)。当数据以每毫克蛋白质、每毫克DNA或每个完整睾丸表示时,可溶性鸟苷酸环化酶活性增加。相反,隐睾睾丸中的颗粒性鸟苷酸环化酶活性降低了40%。对从腹腔和阴囊睾丸制备的可溶性酶进行动力学分析,两种酶制剂均产生线性Line-weaver-Burke图,对三磷酸鸟苷具有相同的K(m),但腹腔酶的最大速度高两倍。当将两个睾丸的可溶性鸟苷酸环化酶混合在一起进行测定时,活性是相加的,而不是表现出协同作用或抑制作用。这些实验表明,V(max)的改变不是由于可转移的激活剂或抑制剂。使用免疫细胞化学技术评估发生cGMP代谢改变的细胞类型。阴囊和腹腔睾丸的比较显示,腹腔睾丸在生精小管内侧衬里细胞以及管状成分和间质细胞内表现出增强的cGMP免疫荧光。因此,推断可溶性鸟苷酸环化酶活性和cGMP水平的相关变化发生在隐睾睾丸内仍存活的几种细胞类型中。