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Inducible Coexpression of Connexin37 or Connexin40 with Connexin43 Selectively Affects Intercellular Molecular Transfer

机译:Connexin37或Connexin40的诱导型共表达与Connexin43选择性地影响细胞间分子转移

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摘要

Many tissues express multiple gap junction proteins, or connexins (Cx); for example, Cx43, Cx40, and Cx37 are coexpressed in vascular cells. This study was undertaken to elucidate the consequences of coexpression of Cx40 or Cx37 with Cx43 at different ratios. EcR-293 cells (which endogenously produce Cx43) were transfected with ecdysone-inducible plasmids encoding Cx37 or Cx40. Immmunoblotting showed a ponasterone dose-dependent induction of Cx37 or Cx40 while constant levels of Cx43 were maintained. The coexpressed connexins colocalized at appositional membranes. Double whole-cell patch clamp recordings showed no significant change in total junctional conductances in cells treated with 0, 0.5, or 4 µM ponasterone; however, they did show a diversity of unitary channel sizes consistent with the induced connexin expression. In cells with induced expression of either Cx40 or Cx37, intercellular transfer of microinjected Lucifer yellow was reduced, but transfer of NBD-TMA (2-(4-nitro- 2,1,3-benzoxadiol-7-yl)[aminoethyl]trimethylammonium) was not affected. In cocultures containing uninduced EcR cells together with cells induced to coexpress Cx37 or Cx40, Lucifer yellow transfer was observed only between the cells expressing Cx43 alone. These data show that induced expression of either Cx37 or Cx40 in Cx43- expressing cells can selectively alter the intercellular exchange of some molecules without affecting the transfer of others.
机译:许多组织表达多个间隙结蛋白,或连接蛋白(CX);例如,CX43,CX40和CX37在血管细胞中共表达。本研究旨在阐明CX40或CX37与CX43以不同比例的后施加的后果。将ECR-293细胞(内源性产生CX43)与编码CX37或CX40的蜕皮型诱导型质粒转染。免疫印迹显示出CX37或CX40的痘剂剂量依赖性诱导,而维持CX43的恒定水平。在对象膜上共聚合的共同的连接蛋白。双重全细胞贴片夹具记录显示出用0,0.5或4μmπ覆盆子处理的细胞中总连接导电的显着变化;然而,他们确实显示了与诱导的Connexin表达一致的单一通道尺寸的多样性。在具有CX40或Cx37的诱导表达的细胞中,减少了微内注射荧光素黄的细胞间转移,但转移NBD-TMA(2-(4-硝基-2,1,3-苯并二醇-7-基)[氨基乙基]三甲基铵)没有受到影响。在含有未屈服的ECR细胞的共培养物中,与诱导CX37或CX40的细胞一起,仅在单独表达CX43的细胞之间观察到荧光素黄色转移。这些数据显示CX43-表达细胞中CX37或CX40的诱导表达可以选择性地改变一些分子的细胞间交换而不影响其它的转移。

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