Ultrastructural demonstration of Cx43 gap junctions in induced pluripotent stem cells from human cord blood

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• 作者：Anja Beckmann ; Madline Schubert ; Nadine Hainz…
• 关键词：iPSC ; Cx43 ; Freeze ; fracture replica immunogold labeling
• 刊名：Histochemistry and Cell Biology
• 出版年：2016
• 出版时间：November 2016
• 年：2016
• 卷：146
• 期：5
• 页码：529-537
• 全文大小：1,297 KB
• 刊物类别：Medicine
• 刊物主题：Medicine & Public Health
  Anatomy
  Medicine/Public Health, general
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-119X
• 卷排序：146

文摘

Gap junction proteins are essential for direct intercellular communication but also influence cellular differentiation and migration. The expression of various connexin gap junction proteins has been demonstrated in embryonic stem cells, with Cx43 being the most intensely studied. As Cx43 is the most prominent gap junction protein in the heart, cardiomyocyte-differentiated stem cells have been studied intensely. To date, however, little is known about the expression and the subcellular distribution of Cx43 in undifferentiated stem cells or about the structural arrangement of channels. We, therefore, here investigate expression of Cx43 in undifferentiated human cord-blood-derived induced pluripotent stem cells (hCBiPS2). For this purpose, we carried out quantitative real-time PCR and immunohistochemistry. For analysis of Cx43 ultrastructure and protein assembly, we performed freeze-fracture replica immunogold labeling (FRIL). Cx43 expression was detected at mRNA and protein level in hCBIPS2 cells. For the first time, ultrastructural data are presented on gap junction morphology in induced pluripotent stem (iPS) cells from cord blood: Our FRIL and electron microscopical analysis revealed the occurrence of gap junction plaques in undifferentiated iPS cells. In addition, these gap junctions were shown to contain the gap junction protein Cx43.