The effects of gap junction blockage on neuronal differentiation of human NTera2/clone D1 cells
The effects of gap junction blockage on neuronal differentiation of human NTera2/clone D1 cells
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DOI：
10.1006/exnr.1998.6950
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发表时间：
1999-03-01
影响因子：
5.3
通讯作者：
Naus, CCG
中科院分区：
文献类型：
--
作者：
Bani-Yaghoub, M;Bechberger, JF;Naus, CCG
间隙连接是细胞间通道，其提供小离子和分子（MW < 1200 D）在相邻细胞之间的通过。NTera 2/克隆D1（NTS/D1）细胞是CNS前体细胞，其在用视黄酸（RA）和抗增殖剂处理后分化成NT 2-N神经元。本研究比较了缝隙连接阻断剂18 α-大黄酸（GRA）和甘珀酸（CBX）与无阻断作用的GRA类似物油酸（奥拉）和甘草酸（GZA）的作用。对照和实验培养物均显示RA诱导4周后Cx43蛋白减少。在该时间点，在对照细胞中也观察到细胞角蛋白、波形蛋白和巢蛋白的表达的显著减少，而用阻断剂处理的培养物未显示出任何显著变化。在用阻断剂处理的培养物中，每个视野的MAP 2阳性NT 2-N分化的神经元的平均数量小于对照培养物的7%。NT 2-N细胞Cx43、cytokeratin、vimentin和nestin均为阴性。阻断剂似乎不通过抑制RA信号传导起作用，因为它们的存在不影响视黄酸受体（RAR α和RAR γ）的表达，也不抑制RA介导的基因转录。这些结果一起表明间隙连接的阻断干扰NT 2/D1细胞的神经元分化，（C）1999 Academic Press。
Gap junctions are intercellular channels which provide for the passage of small ions and molecules (MW < 1200 D) among adjacent cells. The NTera2/clone D1 (NTS/D1) cells are CNS precursors which differentiate into NT2-N neurons upon treatment with retinoic acid (RA) and antiproliferative agents. In this study, the effects of gap junction blockers 18 alpha-glycyrrhetinic acid (GRA) and carbenoxolone (CBX) have been compared with those of oleanolic acid (OLA) and glycyrrhizic acid (GZA), GRA analogs with no blocking effects. Both control and experimental cultures showed reduction of Cx43 protein after 4 weeks of RA induction. A major reduction was also observed in expression of cytokeratin, vimentin, and nestin in control cells at this time point while the cultures treated with the blockers did not show any significant change. The average number of MAP2-positive NT2-N differentiated neurons per field of view in the cultures treated with the blockers was less than 7% of that of control cultures. NT2-N cells were negative for Cx43, cytokeratin, vimentin, and nestin. The blockers did not appear to be operating through inhibition of RA signaling, as their presence did not affect the expression of retinoic acid receptors (RAR alpha and RAR gamma) nor did they inhibit RA-mediated gene transcription, These results, together, show that the blockage of gap junctions interferes with neuronal differentiation of NT2/D1 cells, (C) 1999 Academic Press.