Implications of pannexin 1 and pannexin 3 for keratinocyte differentiation
Implications of pannexin 1 and pannexin 3 for keratinocyte differentiation
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DOI：
10.1242/jcs.056093
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发表时间：
2010-04-15
影响因子：
4
通讯作者：
Laird, Dale W.
中科院分区：
文献类型：
--
作者：
Celetti, Steven J.;Cowan, Kyle N.;Laird, Dale W.
Pannexin（Panx）1和Panx 3是与无脊椎动物间隙连接的连接蛋白家族共享一些序列同源性的整合膜蛋白。它们在哺乳动物皮肤中表达。Pannexins已被报道形成功能性机械敏感性单膜通道，但其在调节细胞功能中的重要性知之甚少。在这项研究中，Panx 1和Panx 3检测到表皮的13.5天的胚胎小鼠。与新生小鼠相比，在薄和厚的小鼠皮肤中Panx 1表达较少，而Panx 3表达不变。为了研究pannexins在角质形成细胞分化中的作用，我们采用了大鼠表皮角质形成细胞（REK），它们具有分化为器官型表皮的能力，并将它们改造为过表达Panx 1，Panx 1-GFP或Panx 3。Panx 1或Panx 3的表达导致REKs摄取染料的能力增加，表明形成了细胞表面通道。与单层REKs相比，内源性Panx 1水平保持不变，而70 kDa的Panx 3免疫反应性物种的器官型表皮大大增加。在单层培养中，异位Panx 1和Panx 1-GFP定位于质膜，而Panx 3显示细胞内和质膜配置文件。虽然这两个pannexins减少细胞增殖，只有Panx 1破坏了器官型表皮的架构和显着失调的细胞角蛋白14的表达和定位。此外，异位表达只有Panx 1减少了重要层厚度的器官型表皮。综上所述，Panx 1和Panx 3在哺乳动物表皮中共表达，并且Panx 1的调节在角质形成细胞分化中起关键作用。
Pannexin (Panx) 1 and Panx3 are integral membrane proteins that share some sequence homology with the innexin family of invertebrate gap junctions. They are expressed in mammalian skin. Pannexins have been reported to form functional mechanosensitive single-membrane channels, but their importance in regulating cellular function is poorly understood. In this study, Panx1 and Panx3 were detected in the epidermis of 13.5 day embryonic mice. Compared with newborn mice, there was less Panx1 expression in both thin and thick murine skin, whereas Panx3 expression was unchanged. To investigate the role of pannexins in keratinocyte differentiation, we employed rat epidermal keratinocytes (REKs) that have the capacity to differentiate into organotypic epidermis, and engineered them to overexpress Panx1, Panx1-GFP or Panx3. The expression of Panx1 or Panx3 resulted in the increased ability of REKs to take up dye, suggesting that cell-surface channels were formed. Compared with monolayer REKs, endogenous Panx1 levels remained unchanged, whereas the 70 kDa immunoreactive species of Panx3 was greatly increased in the organotypic epidermis. In monolayer cultures, ectopic Panx1 and Panx1-GFP localized to the plasma membrane, whereas Panx3 displayed both intracellular and plasma-membrane profiles. Although both pannexins reduced cell proliferation, only Panx1 disrupted the architecture of the organotypic epidermis and markedly dysregulated cytokeratin 14 expression and localization. Furthermore, ectopic expression of only Panx1 reduced the vital layer thickness of the organotypic epidermis. In summary, Panx1 and Panx3 are coexpressed in the mammalian epidermis, and the regulation of Panx1 plays a key role in keratinocyte differentiation.