Cell-to-cell communication in a differentiating ovine lens culture system.
Cell-to-cell communication in a differentiating ovine lens culture system.
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DOI：
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发表时间：
1994
影响因子：
4.4
通讯作者：
E. Tenbroek;R. Johnson;C. Louis
中科院分区：
文献类型：
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作者：
E. Tenbroek;R. Johnson;C. Louis
本研究旨在确定绵羊透镜培养物的上皮细胞和分化中的纤维样细胞之间的连接是否与其他组织中的间隙连接一样，表现出被正辛醇抑制的细胞间通讯，并确定这些绵羊透镜细胞培养物是否表达透镜连接蛋白和纤维细胞膜蛋白MP 20和MP 26。方法用荧光黄CH注射细胞以测量细胞间通讯。连接蛋白相关的透镜膜蛋白MP 70、连接蛋白43（Cx43）和连接蛋白46（Cx46）以及膜蛋白MP 20和MP 26的抗体用于免疫荧光标记透镜培养物并探测从透镜培养物分离的膜的Western印迹。结果荧光黄在上皮细胞和分化中的透明细胞中均有细胞间的转移，但被正辛醇抑制。尽管Cx43抗体免疫荧光标记上皮细胞之间的小斑块，但MP70抗体标记大斑块以及分化中的纤维样细胞的小点状区域。有趣的是，Cx43和MP70经常存在于上皮细胞之间的细胞界面处的相同斑块中以及分化中的纤维样细胞上的一些较大斑块中。Cx46和MP70抗体在分化中的纤维样细胞和晚期上皮细胞的膜中标记相同的斑块。用碱性磷酸酶处理后，所有三种连接蛋白蛋白的电泳迁移率被修改。这些分化区域的免疫组织化学染色和来自分化培养物的纯化膜的Western免疫印迹也显示了MP20和MP26的存在。结论：绵羊透镜培养物中不同类型的细胞表现出间隙连接介导的细胞间通讯，这种通讯可能受到一种或多种连接蛋白的影响。
PURPOSE This study was performed to determine whether the junctions between both the epithelial and the differentiating fiber-like cells of ovine lens cultures, like gap junctions in other tissues, exhibit cell-to-cell communication that is inhibited by n-octanol, and to determine whether lens connexins and the fiber cell membrane proteins MP20 and MP26 are expressed by these ovine lens cell cultures. METHODS Cells were injected with Lucifer yellow CH to measure cell-to-cell communication. Antibodies to connexin-related lens membrane protein MP70, connexin 43 (Cx43), and connexin 46 (Cx46) and to membrane proteins MP20 and MP26 were used to immunofluorescently label lens cultures and probe Western blots of membranes isolated from lens cultures. RESULTS Both epithelial cells and differentiating clear cells exhibited cell-to-cell transfer of Lucifer yellow that was inhibited by n-octanol. Although a Cx43 antibody immunofluorescently labeled small plaques between the epithelial cells, an MP70 antibody labeled large plaques as well as small punctate areas of the differentiating fiber-like cells. It is interesting that Cx43 and MP70 were frequently present in the same plaques at cell interfaces between epithelial cells as well as some of the larger plaques on the differentiating fiber-like cells. Cx46 and MP70 antibodies labeled the same plaques in membranes of differentiating fiber-like cells and late-stage epithelial cells. The electrophoretic mobility of all three connexin proteins was modified after treatment with alkaline phosphatase. Immunohistochemical staining of these differentiating regions and Western immunoblotting of purified membranes derived from differentiated cultures also showed the presence of MP20 and MP26. CONCLUSIONS The different cell types in the ovine lens culture exhibit gap junction-mediated cell-to-cell communication that is likely effected by one or more of the connexin proteins.