Lentivirus-mediated transduction of connexin cDNAs shows level- and isoform-specific alterations in insulin secretion of primary pancreatic β-cells

被引:37
作者
Caton, D [1 ]
Calabrese, A
Mas, C
Serre-Beinier, V
Charollais, A
Caille, D
Zufferey, R
Trono, D
Meda, P
机构
[1] Univ Geneva, Sch Med, Dept Morphol, CH-1211 Geneva 4, Switzerland
[2] Univ Geneva, Sch Med, Dept Genet & Microbiol, CH-1211 Geneva 4, Switzerland
关键词
Cx32; Cx36; Cx43; gap junctions; lentiviral vectors; pancreatic beta-cells;
D O I
10.1242/jcs.00442
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
We have generated novel lentiviral vectors to integrate various connexin cDNAs into primary, non-dividing cells. We have used these vectors to test whether proper control of insulin secretion depends on a specific connexin isoform and/or on its level of expression. We have observed that transduced connexin32, connexin36 and connexin43 were expressed by primary adult beta-cells at membrane interfaces, were packed into typical gap junction plaques and formed functional channels that allowed a variable coupling, depending on the type and level of connexin expressed. The infected cells spontaneously reaggregated into three-dimensional pseudo-islet organs that could be maintained in culture. We have found that pseudo-islets made by cells transduced with either GFP- or connexin43-expressing lentivirus released insulin in response to various secretagogues similarly to controls. By contrast, pseudo-islets made by cells expressing connexin32, a connexin exogenous to pancreatic islets, or over-expressing connexin36, the endogenous islet connexin, featured a marked decrease in the secretory response to glucose. The data show: (1) that lentiviral vectors allow stable modulation of various connexin in primary, non-proliferating cells; (2) that specific connexin isoforms affect insulin secretion differently; and (3) that adequate levels of coupling via connexin36 channels are required for proper beta-cell function.
引用
收藏
页码:2285 / 2294
页数:10
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