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来自Cx46和Cx50基因敲除小鼠晶状体纤维细胞的结构和免疫细胞化学改变

Structural and immunocytochemical alterations in eye lens fiber cells from Cx46 and Cx50 knockout mice.

作者信息

Dunia Irene, Cibert Christian, Gong Xiaohua, Xia Chun-hong, Recouvreur Michel, Levy Essy, Kumar Nalin, Bloemendal Hans, Benedetti E Lucio

机构信息

Biologie Cellulaire, Institut Jacques Monod, CNRS, Universités Paris 6-Paris 7, 2, place Jussieu, F-75251 Paris Cedex 5, France.

出版信息

Eur J Cell Biol. 2006 Aug;85(8):729-52. doi: 10.1016/j.ejcb.2006.03.006. Epub 2006 Jun 5.

Abstract

In the current study we describe the changes of overall organization of lens fiber cells in connexin 46 (Cx46) and connexin 50 (Cx50) knockout mice. Morphometric analyses and the application of immunocytochemical techniques revealed that in Cx46 knockout lens (Cx46 -/-), where Cx50 is expressed alone, the postnatal differentiation of secondary fiber cells proceeds faster and is characterized by an increased number of smaller fiber cells. Conversely, in Cx50 knockout mice (Cx50 -/-), the lenticular mass is considerably reduced and characterized by a small number of fiber cells added during the postnatal period. The process of terminal differentiation was impaired and generated larger fiber cells still possessing cytoplasmic organelles. Freeze-fracture and fracture labeling revealed that the junctional assembly, packing organization and topographic interactions between connexons and MP26 differed when Cx46 and Cx50 were co-assembled in the wild-type or expressed separately in the two distinct knockout phenotypes. Filipin cytochemistry provided indirect evidence that Cx46 and Cx50 expressed alone are recruited into different lipid environments. Our results represent the structural proof that interaction of connexins and MP26 contributes to the overall organization of the fiber cells.

摘要

在本研究中,我们描述了连接蛋白46(Cx46)和连接蛋白50(Cx50)基因敲除小鼠晶状体纤维细胞整体组织结构的变化。形态计量分析和免疫细胞化学技术的应用显示,在仅表达Cx50的Cx46基因敲除晶状体(Cx46 -/-)中,次级纤维细胞的出生后分化进行得更快,其特征是较小的纤维细胞数量增加。相反,在Cx50基因敲除小鼠(Cx50 -/-)中,晶状体质量显著降低,其特征是出生后添加的纤维细胞数量较少。终末分化过程受到损害,产生的较大纤维细胞仍含有细胞质细胞器。冷冻断裂和断裂标记显示,当Cx46和Cx50在野生型中共同组装或在两种不同的基因敲除表型中单独表达时,连接子之间的连接组装、堆积组织以及连接子与MP26之间的拓扑相互作用有所不同。菲律宾菌素细胞化学提供了间接证据,表明单独表达的Cx46和Cx50被募集到不同的脂质环境中。我们的结果提供了结构证据,证明连接蛋白与MP26的相互作用有助于纤维细胞的整体组织。

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