CFTR-dependent membrane insertion is linked to stimulation of the CFTR chloride conductance
1996
We examined the relation between Cl current (I Cl ) stimulation and cell membrane capacitance (C m ) when cystic fibrosis transmembrane conductance regulator (CFTR) was expressed in Xenopus oocytes. I Cl and C m increased in parallel when oocytes expressing CFTR were stimulated by forskolin (10 μM) and 3-isobutyl-1-methylxanthine (1 mM). The adenosine 3',5'-cyclic monophosphate (cAMP)-induced increase in surface area detected by C m was confirmed by morphometry in the same oocytes used for the electrical recordings. These increases in I Cl and C m were reversible and were absent from control oocytes not injected with CFTR cRNA. The time to reach peak I Cl lagged slightly behind the peak in C m . I Cl 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 I Cl and C m , and the kinetics of I Cl and C m stimulation were more rapid the larger the magnitude of the stimulated current. The C m -I Cl 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 I Cl 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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