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连接蛋白基因在小鼠视网膜中的表达模式。

Expression patterns of connexin genes in mouse retina.

作者信息

Güldenagel M, Söhl G, Plum A, Traub O, Teubner B, Weiler R, Willecke K

机构信息

Institut für Genetik, Abteilung Molekulargenetik, Universität Bonn, 53117 Bonn, Germany.

出版信息

J Comp Neurol. 2000 Sep 18;425(2):193-201.

Abstract

To analyze the molecular basis of gap junctional communication in mouse retina, we examined the expression pattern of the following 13 connexin (Cx) genes: Cx26, Cx30, Cx30.3, Cx31, Cx31.1, Cx32, Cx36, Cx37, Cx40, Cx43, Cx45, Cx46, and Cx50. By using reverse transcriptase-polymerase chain reactions with primer oligonucleotides to murine connexin genes, we detected mRNAs of Cx26, Cx31, Cx32, Cx36, Cx37, Cx40, Cx43, Cx45, and Cx50. Retinae from heterozygous mice with targeted replacement of most of the Cx45 open reading frame by a lacZ reporter gene showed Cx45 promoter activity in somata of the ganglion cell layer and the inner nuclear layer. Immunoblot and immunofluorescence analyses with antibodies generated to murine connexin epitopes revealed the presence of Cx36, Cx37, Cx43, and Cx45 proteins: The outer and inner plexiform layer were immunopositive for Cx36 and Cx45. Cx37 immunoreactivity was found in blood vessels of the inner retina. Cx43 immunolabeling was detected in the ganglion cell layer and nerve fiber layer where it was largely colocalized with immunostaining of glial fibrillary acidic protein suggesting that Cx43-positive cells could be of glial origin. No Cx26 protein was detected in retina by using Cx26 antibodies for immunoblot analyses or confocal microscopy. Furthermore, comparative immunofluorescence analyses of retinae from mice deficient for Cx31, Cx32, or Cx40 with retinae of wild-type mice revealed no specific immunostaining. Our results demonstrate regional specificity in expression of connexin genes in mouse retina and, thus, provide a basis for future assignments of functional defects in connexin-deficient mice to cells in different regions of the retina.

摘要

为了分析小鼠视网膜中缝隙连接通讯的分子基础,我们检测了以下13种连接蛋白(Cx)基因的表达模式:Cx26、Cx30、Cx30.3、Cx31、Cx31.1、Cx32、Cx36、Cx37、Cx40、Cx43、Cx45、Cx46和Cx50。通过使用针对小鼠连接蛋白基因的引物寡核苷酸进行逆转录聚合酶链反应,我们检测到了Cx26、Cx31、Cx32、Cx36、Cx37、Cx40、Cx43、Cx45和Cx50的mRNA。来自杂合小鼠的视网膜,其大部分Cx45开放阅读框被lacZ报告基因靶向替换,在神经节细胞层和内核层的胞体中显示出Cx45启动子活性。用针对小鼠连接蛋白表位产生的抗体进行免疫印迹和免疫荧光分析,揭示了Cx36、Cx37、Cx43和Cx45蛋白的存在:外网状层和内网状层对Cx36和Cx45呈免疫阳性。在视网膜内层的血管中发现了Cx37免疫反应性。在神经节细胞层和神经纤维层检测到Cx43免疫标记,在很大程度上它与胶质纤维酸性蛋白的免疫染色共定位,这表明Cx43阳性细胞可能起源于胶质细胞。通过使用Cx26抗体进行免疫印迹分析或共聚焦显微镜检查,在视网膜中未检测到Cx26蛋白。此外,对Cx31、Cx32或Cx40缺陷小鼠的视网膜与野生型小鼠的视网膜进行比较免疫荧光分析,未发现特异性免疫染色。我们的结果证明了小鼠视网膜中连接蛋白基因表达的区域特异性,从而为未来将连接蛋白缺陷小鼠的功能缺陷归因于视网膜不同区域的细胞提供了基础。

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