Mizuki J, Masumoto N, Tahara M, Fukami K, Mammoto A, Tasaka K, Miyake A
Department of Obstetrics and Gynaecology, Osaka University Medical School, Japan.
J Endocrinol. 1995 Feb;144(2):193-200. doi: 10.1677/joe.0.1440193.
These studies were undertaken to characterize the exocytotic changes in purified gonadotropes by three-dimensional imaging using scanning electron microscopy. Rat gonadotropes were purified using a fluorescence-activated cell sorter and an argon laser treatment system. The purified gonadotropes were stimulated with GnRH under various conditions and fixed for scanning electron microscopy. After the GnRH stimulation, many 'hole' structures (diameter 0.1-0.5 micron) were observed on the cell surface, and notably the population of cells with 10 or more holes was clearly increased. The pattern of the time-course of the changes in this population was perfectly consistent with the LH secretory profile of pituitary cells, and their formation of the cells with 10 or more holes was completely inhibited by pretreatment with a GnRH antagonist. Our data suggest that the hole structure represents an exocytotic opening site and that regulated exocytosis in purified gonadotropes can be evaluated by scanning electron microscopy. This method may be widely applicable to other endocrine cells.
这些研究旨在通过使用扫描电子显微镜的三维成像来表征纯化的促性腺激素细胞中的胞吐变化。使用荧光激活细胞分选仪和氩激光处理系统纯化大鼠促性腺激素细胞。在各种条件下用促性腺激素释放激素（GnRH）刺激纯化的促性腺激素细胞，并固定用于扫描电子显微镜检查。GnRH刺激后，在细胞表面观察到许多“孔”结构（直径0.1 - 0.5微米），特别是具有10个或更多孔的细胞群体明显增加。该群体变化的时间进程模式与垂体细胞的促黄体生成素（LH）分泌曲线完全一致，并且用GnRH拮抗剂预处理可完全抑制具有10个或更多孔的细胞的形成。我们的数据表明，孔结构代表一个胞吐开放位点，并且纯化的促性腺激素细胞中的调节性胞吐作用可以通过扫描电子显微镜进行评估。这种方法可能广泛适用于其他内分泌细胞。
