Gap junction formation between cultured embryonic lens cells is inhibited by antibody to N-cadherin
Gap junction formation between cultured embryonic lens cells is inhibited by antibody to N-cadherin
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DOI：
10.1006/dbio.1996.0237
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发表时间：
1996-10-10
影响因子：
2.7
通讯作者：
Johnson, RG
中科院分区：
文献类型：
--
作者：
Frenzel, EM;Johnson, RG
由缝隙连接介导的细胞间通讯对于无血管透镜中的组织稳态是重要的，并且在纤维细胞分化和透镜发育期间在纤维细胞之间形成广泛的缝隙连接区域。我们研究了钙依赖性细胞粘附分子N-钙粘蛋白在纤维细胞间缝隙连接形成过程中的作用。从鸡胚透镜培养物中分离出具有分化纤维细胞特征的多细胞结构Lentoids，随后配对以提供纤维细胞相互作用的体外模型。通过观察荧光染料（钙黄绿素或荧光黄）在长达48小时的时间过程中从晶状体到晶状体的转移来监测成对晶状体细胞之间的间隙连接形成。向培养基中加入Fab片段后，晶状体之间的染料转移受到抑制（100-622 μ g/ml）的对N-钙粘蛋白特异的单克隆抗体，并且还通过减少孵育培养基中的细胞外钙。然而，将抗体的Fab片段（100-1500 μ g/ml）添加到纤维细胞特异性整合膜蛋白MIP中，并没有改变晶状体之间染料转移的时间过程和程度。我们的研究结果，使用培养的胚胎细胞，从细胞系和转染细胞的先前研究扩展。我们的结论是，钙粘蛋白的相互作用，促进胚胎透镜纤维细胞之间的缝隙连接的形成，通过稳定膜并置和/或通过产生细胞内信号（S）。(C)出版社：Academic Press，Inc.
Intercellular communication mediated by gap junctions is important for tissue homeostasis in the avascular lens, and extensive areas of gap junctions form between fiber cells during fiber cell differentiation and lens development. We examined the role of the calcium-dependent cell adhesion molecule, N-cadherin, in the process of gap junction formation between fiber cells. Lentoids, multicellular structures with characteristics of differentiated fiber cells, were isolated from embryonic chick lens cultures and subsequently paired to provide an in vitro model of fiber cell interactions. Gap junction formation between cells of paired lentoids was monitored by observing the lentoid-to-lentoid transfer of fluorescent dyes, either calcein or Lucifer yellow, over a time course of up to 48 hr. Dye transfer between lentoids was inhibited upon the addition to the medium of Fab fragments (100-622 mu g/ml) of a monoclonal antibody specific for N-cadherin, and also by the reduction of extracellular calcium in the incubation medium. However, the addition of Fab fragments (100-1500 mu g/ml) of an antibody to a fiber-cell-specific integral membrane protein, MIP, did not change the time course nor extent of dye transfer between lentoids. Our results, using cultured embryonic cells, extend those from previous studies with cell lines and transfected cells. We conclude that cadherin interactions facilitate the formation of gap junctions between embryonic lens fiber cells, by the stabilization of membrane appositions and/or by the generation of an intracellular signal(s). (C) 1996 Academic Press, Inc.