Yoshikawa Shunichi, Vila Alejandro, Segelken Jasmin, Lin Ya-Ping, Mitchell Cheryl K, Nguyen Duc, O'Brien John
Richard S. Ruiz M.D., Department of Ophthalmology and Visual Science, University of Texas Health Science Center at Houston, Houston, Texas.
University of Texas Graduate School of Biomedical Sciences at Houston, Houston, Texas.
Dev Neurobiol. 2017 May;77(5):548-561. doi: 10.1002/dneu.22418. Epub 2016 Jul 26.
In the mammalian central nervous system, a remarkably small number of connexins is used in electrical synapses, with the majority formed from Cx36. A larger number has been detected in teleosts, with some seeming to serve restricted roles. Here, we report the discovery of a new connexin expressed in the zebrafish lens and a limited set of neurons. Zebrafish cx79.8 (gja8a), previously annotated incorrectly as cx50.5 based on a partial cDNA sequence, is a homologue of mammalian Cx50 (Gja8). We examined its expression through transgenic promoter-reporter constructs, in situ hybridization, and immunolabeling, and examined regulation of coupling in transfected HeLa cells. cx79.8 was expressed most strongly in the lens, but expression was also found in several groups of neurons in the cerebellum and related areas at the midbrain-hindbrain boundary, in cone photoreceptors, and in neurons in the retinal inner nuclear and ganglion cell layers. Labeling in the retina with antibodies against two C-terminal epitopes revealed numerous small punctate spots in the inner plexiform layer and along the somata of cones. Abundant gap junctions were labeled in the outer 1/3 of the lens, but were absent from the center, suggesting that the epitopes or the entire protein was absent from the center. Cx79.8 tracer coupling was strongly regulated by phosphorylation, and was extremely low in control conditions in HeLa cells due to protein phosphatase 2A activity. These properties allow coupling to be strongly restricted in situ, a frequently observed property for electrical synapses. © 2016 Wiley Periodicals, Inc. Develop Neurobiol 77: 548-561, 2017.
在哺乳动物的中枢神经系统中,用于电突触的连接蛋白数量非常少,其中大多数由Cx36形成。在硬骨鱼中检测到的连接蛋白数量较多,其中一些似乎发挥着有限的作用。在这里,我们报告了在斑马鱼晶状体和一组有限的神经元中表达的一种新连接蛋白的发现。斑马鱼cx79.8(gja8a),此前基于部分cDNA序列被错误注释为cx50.5,是哺乳动物Cx50(Gja8)的同源物。我们通过转基因启动子 - 报告基因构建体、原位杂交和免疫标记研究了其表达,并在转染的HeLa细胞中研究了偶联的调节。cx79.8在晶状体中表达最强,但在小脑的几组神经元以及中脑 - 后脑边界的相关区域、视锥光感受器以及视网膜内核层和神经节细胞层的神经元中也有表达。用针对两个C末端表位的抗体对视网膜进行标记,在内网状层和视锥细胞体周围发现了许多小的点状斑点。晶状体外部1/3有大量间隙连接被标记,但中心没有,这表明中心没有表位或整个蛋白质。Cx79.8示踪剂偶联受磷酸化强烈调节,并且由于蛋白磷酸酶2A的活性,在HeLa细胞的对照条件下极低。这些特性使得偶联在原位受到强烈限制,这是电突触经常观察到的特性。©2016威利期刊公司。《发育神经生物学》77:548 - 56,2017。