Chang Q, Gonzalez M, Pinter M J, Balice-Gordon R J
Department of Neuroscience, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104-6074, USA.
J Neurosci. 1999 Dec 15;19(24):10813-28. doi: 10.1523/JNEUROSCI.19-24-10813.1999.
Interneuronal gap junctional coupling is a hallmark of neural development whose functional significance is poorly understood. We have characterized the extent of electrical coupling and dye coupling and patterns of gap junction protein expression in lumbar spinal motor neurons of neonatal rats. Intracellular recordings showed that neonatal motor neurons are transiently electrically coupled and that electrical coupling is reversibly abolished by halothane, a gap junction blocker. Iontophoretic injection of Neurobiotin, a low molecular weight compound that passes across most gap junctions, into single motor neurons resulted in clusters of many labeled motor neurons at postnatal day 0 (P0)-P2, and single labeled motor neurons after P7. The compact distribution of dye-labeled motor neurons suggested that, after birth, gap junctional coupling is spatially restricted. RT-PCR, in situ hybridization, and immunostaining showed that motor neurons express five connexins, Cx36, Cx37, Cx40, Cx43, and Cx45, a repertoire distinct from that expressed by other neurons or glia. Although all five connexins are widely expressed among motor neurons in embryonic and neonatal life, Cx36, Cx37, and Cx43 continue to be expressed in many adult motor neurons, and expression of Cx45, and in particular Cx40, decreases after birth. The disappearance of electrical and dye coupling despite the persistent expression of several gap junction proteins suggests that gap junctional communication among motor neurons may be modulated by mechanisms that affect gap junction assembly, permeability, or open state.
神经元间缝隙连接耦合是神经发育的一个标志,但其功能意义仍知之甚少。我们已经对新生大鼠腰段脊髓运动神经元的电耦合程度、染料耦合以及缝隙连接蛋白的表达模式进行了表征。细胞内记录显示,新生运动神经元短暂地进行电耦合,并且电耦合可被缝隙连接阻滞剂氟烷可逆性阻断。将Neurobiotin(一种能穿过大多数缝隙连接的低分子量化合物)离子电渗注入单个运动神经元,在出生后第0天(P0)-P2导致许多标记的运动神经元成簇出现,而在P7之后则出现单个标记的运动神经元。染料标记的运动神经元的紧密分布表明,出生后缝隙连接耦合在空间上受到限制。逆转录-聚合酶链反应(RT-PCR)、原位杂交和免疫染色显示,运动神经元表达五种连接蛋白,即Cx36、Cx37、Cx40、Cx43和Cx45,这一组合与其他神经元或神经胶质细胞所表达的不同。尽管所有这五种连接蛋白在胚胎期和新生期的运动神经元中广泛表达,但Cx36、Cx37和Cx43在许多成年运动神经元中仍持续表达,而Cx45,尤其是Cx40的表达在出生后下降。尽管几种缝隙连接蛋白持续表达,但电耦合和染料耦合消失,这表明运动神经元之间的缝隙连接通讯可能受到影响缝隙连接组装、通透性或开放状态的机制的调节。