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AJP - Cell Physiology, Vol 271, Issue 6 C1887-C1894, Copyright © 1996 by American Physiological Society
ARTICLES |
A. Takahashi, S. C. Watkins, M. Howard and R. A. Frizzell
Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, Pennsylvania 15261, USA.
We examined the relation between Cl current (Icl) stimulation and cell membrane capacitance (Cm) when cystic fibrosis transmembrane conductance regulator (CFTR) was expressed in Xenopus oocytes. ICl and Cm increased in parallel when oocytes expressing CFTR were stimulated by forskolin (10 microM) and 3-isobutyl-1-methylxanthine (1 mM). The adenosine 3',5'-cyclic monophosphate (cAMP)-induced increase in surface area detected by Cm was confirmed by morphometry in the same oocytes used for the electrical recordings. These increases in ICl and Cm were reversible and were absent from control oocytes not injected with CFTR cRNA. The time to reach peak ICl lagged slightly behind the peak in Cm. ICl was varied by altering CFTR expression level or agonist dose or by expressing different CFTR mutants. In all cases, there was a close correlation between ICl and Cm, and the kinetics of ICl and Cm stimulation were more rapid the larger the magnitude of the stimulated current. The Cm-ICl relation for wild-type CFTR saturated, consistent with a limited capacity of cells to increase their surface area. These results indicate that stimulation of the CFTR ICl is linked closely to increases in membrane area. This suggests that CFTR is present in the membrane vesicles whose insertion is stimulated by cAMP. The contents of these vesicles may provide a link between activation of CFTR and its cAMP-dependent regulation of other channels.
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