Caveolae and Endoplasmic Reticulum: Immunofluorescence Microscopy and Time-Lapse Analysis.

DOI Web Site 被引用文献4件 参考文献17件
  • Kogo Hiroshi
    Department of Anatomy and Cell Biology, Gunma University School of Medicine
  • Shioya Mariko
    Department of Anatomy and Cell Biology, Gunma University School of Medicine
  • Takahashi Yukiko
    Department of Anatomy and Cell Biology, Gunma University School of Medicine
  • Fujimoto Toyoshi
    Department of Anatomy and Cell Biology, Gunma University School of Medicine

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Caveolae have been hypothesized to be involved in Ca2+ signaling. By electron microscopy, caveolae were observed to be apposed to the endoplasmic reticulum (ER), which is a major intracellular Ca2+ pool. In the present study, we examined the relationship between caveolae and the ER when the distribution of the latter was changed by depolymerization of microtubules. Double immunofluorescence microscopy for detection of caveolin and the ER antigens, and time-lapse observation of green fluorescent protein (GFP) -tagged caveolin were employed. In normal human fibroblasts and PtK2 cells, the ER was seen as a network extending throughout the cytoplasm, and most caveolin occurred in patches along the cell edge. When microtubules were depolymerized by Colcemid or nocodazole, the ER became retracted from the cell periphery and aggregated around the nucleus; in the same cells, caveolin was not seen along the cell edge, but was aligned along the edge of the retracted ER. By time-lapse analysis, GFPcaveolin expressed in PtK2 cells was observed to move from the cell periphery toward the cell center in Colcemid-treated cells. The result shows that the apposition of caveolae and the ER is maintained even after the gross distributional change, and suggests a mechanical linkage between the two organelles.

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