Three-dimensional, Computer-tomographic Analysis of Membrane Proteins (TrkA, caveolin, clathrin) in PC12 Cells

DOI NDLデジタルコレクション Web Site 被引用文献3件 参考文献45件
  • Nishida Tomoki
    Division of Applied Biology, Kyoto Institute of Technology
  • Arii Tatsuo
    Center for Brain Experiment, National Institute for Physiological Sciences
  • Takaoka Akio
    Research Center for Ultra-High Voltage Electron Microscopy, Osaka University
  • Yoshimura Ryoichi
    Division of Applied Biology, Kyoto Institute of Technology
  • Endo Yasuhisa
    Division of Applied Biology, Kyoto Institute of Technology

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Signaling of nerve growth factor (NGF) and its receptor (TrkA) promotes neuronal differentiation, synapse formation and survival. It has been known that the complex of NGF and TrkA is internalized into the cytoplasm and transported for further signal transduction, but the ultrastructural information of this process is virtually unknown. In order to clarify the relationship between the internalization of TrkA and the membrane-associated proteins (caveolin and clathrin), the localization and three-dimensional structures of those proteins were examined with computer tomography of high voltage electron microscopy in PC12 cells. TrkA immunoreactivity was found only at definite areas in the plasma membrane, as ring and cluster structures. Its 3D image indicated that those cluster structures contained small pits, which did not appear to be typical caveolae in size and shape. 3D images of clathrin and caveolin-1 immunoreactivities indicated that the formation of those small pits was associated with clathrin, but not with caveolin-1. Caveolin-1 immunoreactivity was found as a mesh-like structure just beneath the plasma membrane. These results suggest that clathrin rather than caveolin is mainly involved in the process of TrkA internalization, at least in differentiated PC12 cells.<br>

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