Pannexin-1 channels show distinct morphology and no gap junction characteristics in mammalian cells

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• 作者：Anja Beckmann ; Alexander Grissmer ; Elmar Krause…
• 关键词：Gap junction ; Pannexin ; Electron microscopy ; Freeze ; fracture replica immunolabeling ; FRIL
• 刊名：Cell and Tissue Research
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：363
• 期：3
• 页码：751-763
• 全文大小：2,998 KB
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• 作者单位：Anja Beckmann (1)
  Alexander Grissmer (1)
  Elmar Krause (2)
  Thomas Tschernig (1)
  Carola Meier (1)
  
  1. Department of Anatomy and Cell Biology, Saarland University, Building 61, 66421, Homburg/Saar, Germany
  2. Department of Physiology, Saarland University, 66421, Homburg/Saar, Germany
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biomedicine
  Human Genetics
  Proteomics
  Molecular Medicine
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-0878

文摘

Pannexins (Panx) are proteins with a similar membrane topology to connexins, the integral membrane protein of gap junctions. Panx1 channels are generally of major importance in a large number of system and cellular processes and their function has been thoroughly characterized. In contrast, little is known about channel structure and subcellular distribution. We therefore determine the subcellular localization of Panx1 channels in cultured cells and aim at the identification of channel morphology in vitro. Using freeze-fracture replica immunolabeling on EYFP-Panx1-overexpressing HEK 293 cells, large particles were identified in plasma membranes, which were immunogold-labeled using either GFP or Panx1 antibodies. There was no labeling or particles in the nuclear membranes of these cells, pointing to plasma membrane localization of Panx1-EYFP channels. The assembly of particles was irregular, this being in contrast to the regular pattern of gap junctions. The fact that no counterparts were identified on apposing cells, which would have been indicative of intercellular signaling, supported the idea of Panx1 channels within one membrane. Control cells (transfected with EYFP only, non-transfected) were devoid of both particles and immunogold labeling. Altogether, this study provides the first demonstration of Panx1 channel morphology and assembly in intact cells. The identification of Panx1 channels as large particles within the plasma membrane provides the knowledge required to enable recognition of Panx1 channels in tissues in future studies. Thus, these results open up new avenues for the detailed analysis of the subcellular localization of Panx1 and of its nearest neighbors such as purinergic receptors in vivo.