Theis Martin, Mas Christophe, Döring Britta, Degen Joachim, Brink Christopher, Caille Dorothée, Charollais Anne, Krüger Olaf, Plum Achim, Nepote Virginie, Herrera Pedro, Meda Paolo, Willecke Klaus
Institut für Genetik, Abteilung Molekulargenetik, Universität Bonn, D-53117 Bonn, Germany.
Exp Cell Res. 2004 Mar 10;294(1):18-29. doi: 10.1016/j.yexcr.2003.09.031.
Transcripts of three connexin isoforms (Cx36, Cx43 and Cx45) have been reported in rodent pancreatic islets, but the precise distribution of the cognate proteins is still unknown. We determined expression of Cx36 in a cell-autonomous manner using mice with a targeted replacement of the Cx36 coding region by a lacZ reporter gene. For cell-autonomous monitoring of Cx43 expression, we used the Cre/loxP system: Mice carrying the Cx43 coding region flanked by loxP sites (floxed) also carried an embedded lacZ gene that is activated after Cre-mediated recombination in cells with transcriptional activity of the Cx43 gene. Deletion of the Cx43 coding region in beta-cells did not result in the activation of the embedded lacZ reporter gene. Instead, Cx43 expression was found in endothelial cells of the islets of Langerhans in mice with endothelium-specific deletion. Ubiquitous deletion of Cx43 led to a similar endothelial lacZ expression, but again, activity of the reporter gene was not detected in beta-cells. Mice with targeted replacement of the Cx45 coding region by lacZ showed a vascular expression similar to Cx43. The data show that native insulin-producing cells express a connexin isoform (Cx36) which differs from those (Cx43 and Cx45) expressed by vascular islet cells.
在啮齿动物胰岛中已报道了三种连接蛋白亚型(Cx36、Cx43和Cx45)的转录本,但相关蛋白的精确分布仍不清楚。我们使用通过lacZ报告基因靶向替换Cx36编码区的小鼠,以细胞自主方式确定Cx36的表达。为了对Cx43表达进行细胞自主监测,我们使用了Cre/loxP系统:携带两侧有loxP位点(floxed)的Cx43编码区的小鼠还携带一个嵌入的lacZ基因,该基因在具有Cx43基因转录活性的细胞中经Cre介导的重组后被激活。β细胞中Cx43编码区的缺失并未导致嵌入的lacZ报告基因的激活。相反,在内皮细胞特异性缺失的小鼠的胰岛内皮细胞中发现了Cx43表达。Cx43的普遍缺失导致类似的内皮细胞lacZ表达,但同样,在β细胞中未检测到报告基因的活性。用lacZ靶向替换Cx45编码区的小鼠表现出与Cx43相似的血管表达。数据表明,天然胰岛素产生细胞表达一种与胰岛血管细胞表达的连接蛋白亚型(Cx43和Cx45)不同的连接蛋白亚型(Cx36)。