The perforated patch clamp recording in cardiac myocytes using
cation-selective ionophore, gramicidin.
Y., Tajima, Ono K. and Akaike N.
Department of Physiology, Kyushu University Faculty of Medicine,
Fukuoka 812-82, Japan, TEL 81-92-641-1151 ext 3336, FAX 81-92-633
-6748
APStracts 3:0071C, 1996.
Whole-cell currents were recorded in single myocytes dissociated from
guinea-pig ventricles by the patch clamp technique. The addition of
0.1 mg/ml gramicidin D, a cation selective ionophore, into the
pipette solution induced a gradual, spontaneous perforation of the
patch membrane under a conventional cell-attached configuration. The
access resistance, measured at around 12 min after forming a giga-ohm
seal, was 9.2 +/- 1.5 M[omega] (n = 12) . The perforated patch
membrane exhibited ionic selectivity for various monovalent cations
with a relative order of Cs+ (1.11) > K+ (1.0) > Na+ (0.65)
>> Tris+( nearly equal to 0), but was not permeable for Cl
-. Under the gramicidin-perforated patch recording configuration, the
cells showed the same typical electrophysiological properties for
ventricular cells as reported so far. The intracellular Cl-
concentration ([Cl-]i), estimated from the reversal potential of the
catecholamine-induced Cl- current (ICl), was 36.3 +/- 2.9 mM (n =
17). We thus conclude that gramicidin perforated patch recording mode
provides a useful tool for recording the ionic currents while
maintaining the [Cl-]i.
Received 11 September 1995; accepted in final form 13 February
1996.
APS Manuscript Number C553-5.
Article publication pending Am. J. Physiol. (Cell Physiology).
ISSN 1080-4757 Copyright 1996 The American Physiological Society.
Published in APStracts on 20 March 96