XB-ART-282
Methods Mol Biol
2006 Jan 01;322:315-29. doi: 10.1007/978-1-59745-000-3_22.
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Studying the mechanosensitivity of voltage-gated channels using oocyte patches.
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The mechanosensitivity of voltage-gated (VG) channels is of biophysical, physiological. and pathophysiological interest. Xenopus oocytes offer a critical advantage for investigating the electrophysiology of recombinant VG channels subjected to membrane stretch, namely, the ability to monitor macroscopic current from membrane patches. High-density channel expression in oocytes makes for macroscopic current in conventional-size, mechanically sturdy patches. With the patch configuration, precisely the same membrane that is voltage-clamped is the membrane subjected to on-off stretch stimuli. With patches, meaningful stretch dose responses are possible. Experimental design should facilitate within-patch comparisons wherever possible. The mechanoresponses of some VG channels depend critically on patch history. Methods for minimizing and coping with interference from endogenous voltage-dependent and stretch-activated endogenous channels are described.
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