Matesic D F, Germak J A, Dupont E, Madhukar B V
Department of Pediatrics and Human Development, Michigan State University, East Lansing 48824.
Neuroendocrinology. 1993 Nov;58(5):485-92. doi: 10.1159/000126581.
Expression of gap junction proteins was studied in the LHRH neuronal cell line, GT1-7, as a first step in defining the signalling mechanisms responsible for the pulsatile secretion of LHRH. GT1-7 cells were found to express a connexin 26-like protein that comigrated with mouse liver connexin 26 and that reacted with connexin 26-specific antibodies on Western blots. Immunofluorescent staining revealed punctate staining in a fraction of the cells, often present at points of apparent contact with neighboring cell bodies or processes. Fluorescence recovery after photobleaching analysis of 5,6-carboxyfluorescein loaded GT1-7 cells showed dye coupling among 20-30% of cells that made contact with other cells, suggesting the presence of functional gap junctions in this cell line.
作为确定负责促黄体生成素释放激素（LHRH）脉冲式分泌的信号传导机制的第一步，我们在LHRH神经元细胞系GT1-7中研究了间隙连接蛋白的表达。发现GT1-7细胞表达一种与小鼠肝脏连接蛋白26共迁移的连接蛋白26样蛋白，并且在蛋白质免疫印迹上与连接蛋白26特异性抗体发生反应。免疫荧光染色显示在一部分细胞中存在点状染色，这些细胞通常出现在与相邻细胞体或突起明显接触的部位。对装载了5,6-羧基荧光素的GT1-7细胞进行光漂白后的荧光恢复分析表明，在与其他细胞接触的20%-30%的细胞之间存在染料偶联，这表明该细胞系中存在功能性间隙连接。
