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Journal of Electron Microscopy Advance Access published online on June 23, 2006
Journal of Electron Microscopy, doi:10.1093/jmicro/dfl017
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© The Author 2006. Published by Oxford University Press on behalf of Japanese Society of Microscopy. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received December 29, 2005
Accepted May 22, 2006

Article

The relationship between intramembranous particles and aquaporin molecules in the plasma membranes of normal rat skeletal muscles: a fracture-label study

Seiji Shibuya 1 *, Yoshihiro Wakayama 1, Masahiko Inoue 1, Hiroko Kojima 1, and Hiroaki Oniki 2

1 Department of Neurology, Showa University Fujigaoka Hospital, 1-30 Fujigaoka, Aoba-Ku, Yokohama 227-8501, Japan
2 Electron Microscopy Laboratory, Showa University Fujigaoka Hospital, 1-30 Fujigaoka, Aoba-Ku, Yokohama 227-8501, Japan

* To whom correspondence should be addressed.
Seiji Shibuya, E-mail: seiji-shibuya{at}nifty.com


  Abstract

The plasma membrane of skeletal muscles contains water channels such as aquaporin 4 (AQP4), aquaporin 3 (AQP3) and aquaporin 7 (AQP7). In dehydrated mice, we have recently reported the altered distribution of the aggregations of intramembranous particles (IMPs), such as orthogonal array (a crystal-like structure) and IMP cluster (a rosette-like structure) on the freeze-fractured skeletal muscle plasma membranes. In this fracture-label study, we first tested whether the orthogonal arrays (OAs) were composed of AQP4 in skeletal muscles and further analyzed the relationship between IMPs including IMP clusters and AQP3 molecules. As a result, many of the gold particles indicating AQP4 was associated with OAs (79%) by our fracture-label technique. On the other hand, ~50% of gold particles indicating AQP3 were associated with IMP clusters. Thus we confirmed that the OAs are composed of AQP4 in skeletal muscles, and further demonstrated that some of the IMP clusters are composed of AQP3 and may participate in maintaining osmotic homeostasis in skeletal myofibers. The fracture-label method is useful in investigating the molecular identification of membrane proteins such as AQP3 and AQP4.

Keywords: aquaporin 3; aquaporin 4; intramembranous particle; orthogonal array; skeletal muscle; fracture-label.
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