Redistribution of connexin 43 during cell division.
Redistribution of connexin 43 during cell division.
复制标题
细胞分裂过程中连接蛋白 43 的重新分布。
DOI：
10.1002/cbin.10576
复制
发表时间：
2016
影响因子：
3.9
通讯作者：
Murray,SandraA
中科院分区：
文献类型：
--
作者：
Vanderpuye,OluseyiA;Bell,CherylL;Murray,SandraA
间隙连接通道一旦聚集成间隙连接斑块，就允许细胞之间必需代谢物的交流。据报道，间隙连接斑块在细胞分裂期间从细胞表面丢失。有丝分裂期间间隙连接斑块丢失的机制尚不清楚，但我们假设涉及导致细胞质双膜环形间隙连接囊泡的内吞机制。在这项研究中，在SW-13肾上腺皮质肿瘤细胞中检查了分裂细胞中的间隙连接斑块变化。用免疫荧光法检测内源性间隙连接蛋白连接蛋白43（Cx43），并使用活细胞成像法监测绿色荧光蛋白标记的Cx43（Cx43-GFP）。Hoechst染色鉴定有丝分裂期。在间期，检测到大的缝隙连接斑块，然而，这些斑块的存在减少，而细胞质斑点增加开始与前期。免疫电子显微镜检查证实胞质斑点为Cx43阳性环状间隙连接囊泡。随着子细胞之间的卵裂沟处间隙连接斑块的重新形成，胞质分裂期间环形间隙连接的数量减少。这些数据与有丝分裂过程中间隙连接斑块丢失的机制一致，该机制依赖于导致环形间隙连接囊泡形成的内分泌过程。胞质分裂过程中间隙连接斑块的快速形成表明，当有丝分裂完成时，连接蛋白从环形间隙连接囊泡中回收形成间隙连接斑块的可能性很有趣。
Gap junction channels, once clustered into gap junction plaques, allow communication of essential metabolites between cells. Gap junction plaques have been reported to be lost from the cell surface during cell division. The mechanism involved in this loss of gap junction plaques during mitosis is unclear, but we hypothesize that an endoexocytotic mechanism that results in cytoplasmic double‐membraned annular gap junction vesicles is involved. In this study, gap junction plaque changes in dividing cells were examined in SW‐13 adrenocortical tumor cells. Endogenous gap junction protein, connexin 43 (Cx43), was detected with immunofluorescence, and live cell imaging was used to monitor green fluorescent protein‐tagged Cx43 (Cx43‐GFP). Mitotic stages were identified by Hoechst chromosomal staining. During interphase, large gap junction plaques were detected; however, the presence of these plaques decreased, whereas cytoplasmic puncta increased beginning with prophase. The cytoplasmic puncta were demonstrated with immunoelectron microscopy to be Cx43‐ positive annular gap junction vesicles. As gap junction plaques reformed at cleavage furrows between daughter cells, the number of annular gap junctions decreased during cytokinesis. The data are consistent with the mechanism of gap junction plaque loss during mitosis relying on an endoexocytotic process that results in annular gap junction vesicles formation. The rapid formation of gap junction plaques during cytokinesis points to the intriguing possibility of connexin recycling from annular gap junction vesicles to form gap junction plaques as mitosis is completed.