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Biophys. J. BioFAST: First Published November 16, 2007. doi:10.1529/biophysj.107.117820
© 2007 by the Biophysical Society.

A more recent version of this article appeared on March 1, 2008.
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CHANNELS, RECEPTORS, AND ELECTRICAL SIGNALING

Spatial Distribution of Maxi-anion Channel on Cardiomyocytes Detected by Smart-Patch Technique

Amal K. Dutta 1, Yuri E. Korchev 2, Andrew I. Shevchuk 2, Seiji Hayashi 1, Yasunobu Okada 1* and Ravshan Z Sabirov 1

1 National Institute for Physiological Sciences
2 Imperial College London, MRC Clinical Sciences Center, Faculty of Medicine

* To whom correspondence should be addressed. E-mail: okada{at}nips.ac.jp.

Submitted on July 20, 2007
Revised on August 26, 2007
Accepted on 24 October 2007


  Abstract
Spatial distribution of maxi-anion channels in rat cardiomyocytes were studied by applying the recently developed patch clamp technique under scanning ion conductance microscopy (SICM), called the "smart-patch"technique. In primary-cultured neonatal cells, the channel was found to be unevenly distributed over the cell surface with significantly lower channel activity in cellular extensions compared to the other parts. Local ATP release, detected using a PC12 cell-based biosensor technique, also exhibited a similar pattern. The maxi-anion channel activity could not be detected in freshly isolated adult cardiomyocytes by the conventional patch-clamp with 2-M{Omega} pipettes. However, when fine-tipped 15-20 M{Omega} pipettes were targeted to only Z-line areas, we observed, for the first time, the maxi-anion events. Smart-patching different regions of the cell surface, we found that the channel activity was maximal at the openings of T-tubules and along Z-lines, but was significantly decreased in the scallop crest area. Thus, it is concluded that maxi-anion channels are concentrated at the openings of T-tubules and along Z-lines in adult cardiomyocytes. The present study showed that the "smart-patch" technique provides a powerful method to detect a unitary event of channels which are localized at some specific site in the narrow region.

Key Words: ATP release, cardiomyocyte, maxi-anion channel, smart-patch






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Copyright © 2007 by the Biophysical Society.