REDUCTION OF GAP JUNCTIONAL CONDUCTANCE BY MICROINJECTION OF ANTIBODIES AGAINST THE 27-KDA LIVER GAP JUNCTION POLYPEPTIDE
REDUCTION OF GAP JUNCTIONAL CONDUCTANCE BY MICROINJECTION OF ANTIBODIES AGAINST THE 27-KDA LIVER GAP JUNCTION POLYPEPTIDE
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DOI：
10.1073/pnas.82.8.2412
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发表时间：
1985-01-01
影响因子：
11.1
通讯作者：
BENNETT, MVL
中科院分区：
文献类型：
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作者：
HERTZBERG, EL;SPRAY, DC;BENNETT, MVL
将分离的大鼠肝脏缝隙连接抗体显微注射到培养的偶联细胞中，观察其对缝隙连接电导的影响。新分离的成年大鼠肝细胞和心肌细胞以及新生大鼠颈上神经节神经元在电紧性突触形成的条件下培养，其荧光染料传递和电耦合迅速受到抑制。间接免疫荧光显示抗体可与肝脏质膜斑点状区域结合。通过对大鼠肝脏匀浆、质膜和分离的缝隙连接在NaDodSO4[十二烷基硫酸钠]/聚丙烯酰胺凝胶上的免疫复制分析，缝隙连接的27 kDa[kDa]主要多肽是特异结合的。这种抗体和类似的抗体应该为进一步研究缝隙连接介导的细胞间通讯提供了工具，并表明免疫上相似的多肽包含来自所有3个生殖层的成年哺乳动物细胞中的缝隙连接。
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 were 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 [sodium dodecyl sulfate]/polyacrylamide gels, binding was specific for the 27-kDa [kilodalton] 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 3 germ layers.