Altered beta-adrenergic and muscarinic response of CFTR Cl- current in dialyzed cardiac myocytes.
Autonomic regulation of the cardiac cystic fibrosis transmembrane conductance regulator (CFTR) Cl- current was studied in isolated guinea pig ventricular myocytes using various configurations of the whole cell patch-clamp technique. When currents were recorded using the conventional patch-clamp technique, it was possible to continue to activate the Cl- current on repeated exposure to isoproterenol (Iso) for up to 60 min after initiating dialysis. However, there was significant rundown of the magnitude of the Cl- current response to the maximally stimulating concentrations of Iso. In addition, the concentration of Iso that produced half-maximal activation of the Cl- current (K1/2) increased with time. Conversely, the K1/2 for acetylcholine inhibition of the Iso-activated current decreased with time. When currents were recorded using the perforated patch-clamp technique, the sensitivity to both beta-adrenergic- and muscarinic-receptor stimulation was stable. Immediately after initiation of dialysis with the conventional patch-clamp technique, the sensitivity to Iso was nearly identical to that determined using the perforated patch-clamp technique. However, the initial sensitivity to muscarinic-receptor activation was significantly greater. These results indicate that cell dialysis associated with conventional patch-clamp techniques not only results in a time-dependent rundown of current amplitude, but it also significantly alters the concentration dependence of beta-adrenergic and muscarinic-receptor regulation of ion channel function.
Zakharov
S I
SI
Department of Physiology and Biophysics, Case Western Reserve University, Cleveland, Ohio 44106, USA.
Harvey
R D
RD
D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D018118:Chloride Channels / Q000187:drug effects / Q000502:physiology*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D003956:Dialysis; D004553:Electric Conductivity; D005260:Female; D006168:Guinea Pigs; D006321:Heart / Q000187:drug effects / Q000502:physiology*; D007545:Isoproterenol / Q000494:pharmacology; D008297:Male; D008565:Membrane Proteins / Q000502:physiology*; D009116:Muscarine / Q000378:metabolism*; D009206:Myocardium / Q000166:cytology; D018408:Patch-Clamp Techniques; D011943:Receptors, Adrenergic, beta / Q000502:physiology*
D018118:Chloride Channels; D008565:Membrane Proteins; D011943:Receptors, Adrenergic, beta; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D009116:Muscarine; D007545:Isoproterenol; D000109:Acetylcholine