Hertzberg E L, Spray D C, Bennett M V
Proc Natl Acad Sci U S A. 1985 Apr;82(8):2412-6. doi: 10.1073/pnas.82.8.2412.
Antibody raised against isolated rat liver gap junctions was microinjected into coupled cells in culture to assess its influence on gap junctional conductance. A rapid inhibition of fluorescent dye transfer and electrical coupling was produced in pairs of freshly dissociated adult rat hepatocytes and myocardial cells as well as in pairs of superior cervical ganglion neurons from neonatal rats cultured under conditions in which electrotonic synapses form. The antibodies have been shown by indirect immunofluorescence to bind to punctate regions of the plasma membrane in liver. By immunoreplica analysis of rat liver homogenates, plasma membranes, and isolated gap junctions resolved on NaDodSO4/polyacrylamide gels, binding was shown to be specific for the 27-kDa major polypeptide of gap junctions. This and similar antibodies should provide a tool for further investigation of the role of cell-cell communication mediated by gap junctions and indicate that immunologically similar polypeptides comprise gap junctions in adult mammalian cells derived from all three germ layers.
将针对分离的大鼠肝脏间隙连接产生的抗体显微注射到培养的偶联细胞中,以评估其对间隙连接电导的影响。在新鲜分离的成年大鼠肝细胞和心肌细胞对中,以及在形成电突触的培养条件下的新生大鼠颈上神经节神经元对中,荧光染料转移和电偶联迅速受到抑制。通过间接免疫荧光显示,这些抗体与肝脏质膜的点状区域结合。通过对大鼠肝脏匀浆、质膜以及在十二烷基硫酸钠/聚丙烯酰胺凝胶上分离的间隙连接进行免疫印迹分析,显示结合对间隙连接的27 kDa主要多肽具有特异性。这种抗体和类似的抗体应该为进一步研究间隙连接介导的细胞间通讯的作用提供一种工具,并表明免疫上相似的多肽构成了来自所有三个胚层的成年哺乳动物细胞中的间隙连接。