Jiang J X, White T W, Goodenough D A
Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Dev Biol. 1995 Apr;168(2):649-61. doi: 10.1006/dbio.1995.1109.
Gap junctions are composed of the connexins, a family of proteins which have been shown to be the structural and functional building blocks of gap junctional intercellular channels. In the chick lens, three members of the connexin (Cx) family have been characterized: Cx 43 has been shown to be a component of interepithelial cell gap junctions, and Cx 45.6 and Cx 56 have been shown to be part of fiber-to-fiber junctions. Early in chick development, gap junctional communication between differentiating lens fibers in the chick loses its sensitivity to blockade by elevated carbon dioxide (CO2). The lens epithelial cells, however, remain CO2-sensitive throughout development. A possible explanation for the change in CO2 sensitivity is that the lens fibers express differentiation-specific connexins with different physiological properties. Using specific antibodies for each of the three lens connexins, we show here that the lens fibers began to express Cx 45.6, and Cx 56 at the developmental stage when lens fibers acquired CO2-insensitivity. However, electron microscopic immunocytochemistry revealed that both Cx 45.6 and Cx 56 were found in interepithelial cell gap junctions, in addition to Cx 43. Conductances between paired Xenopus oocytes injected with Cx 43, Cx 45.6 and Cx 56 mRNAs revealed that all three connexins were CO2-sensitive in this expression system. Taken together, these data ruled out the possibility that the change in CO2 sensitivity observed in vivo could be explained on the basis of the distribution of connexins alone; other parameters of cellular context, such as post-translational processing, must be involved in the observed developmental changes in physiology.
间隙连接由连接蛋白组成,这是一类蛋白质,已被证明是间隙连接细胞间通道的结构和功能构建单元。在鸡晶状体中,连接蛋白(Cx)家族的三个成员已被鉴定:Cx 43已被证明是上皮细胞间间隙连接的一个组成部分,而Cx 45.6和Cx 56已被证明是纤维与纤维连接的一部分。在鸡胚胎发育早期,鸡晶状体中分化的晶状体纤维之间的间隙连接通讯对二氧化碳(CO2)升高引起的阻断失去敏感性。然而,晶状体上皮细胞在整个发育过程中仍对CO2敏感。CO2敏感性变化的一个可能解释是,晶状体纤维表达具有不同生理特性的分化特异性连接蛋白。我们使用针对三种晶状体连接蛋白的特异性抗体,在此表明,在晶状体纤维获得对CO2不敏感的发育阶段,晶状体纤维开始表达Cx 45.6和Cx 56。然而,电子显微镜免疫细胞化学显示,除了Cx 43外,Cx 45.6和Cx 56也存在于上皮细胞间间隙连接中。注射Cx 43、Cx 45.6和Cx 56 mRNA的成对非洲爪蟾卵母细胞之间的电导显示,在这个表达系统中,所有三种连接蛋白都对CO2敏感。综上所述,这些数据排除了仅根据连接蛋白的分布来解释体内观察到的CO2敏感性变化的可能性;细胞环境的其他参数,如翻译后加工,必定参与了观察到的生理发育变化。