title: Mechanisms of GABA(A) receptor blockade by millimolar concentrations of furosemide in isolated rat Purkinje cells.
abstract: The action of diuretic furosemide on the GABA(A) receptor was studied in acutely isolated Purkinje cells using the whole-cell recording and fast application system. Furosemide blocked stationary component of GABA-activated currents in a concentration-dependent manner with IC(50) value > 5 mM at -70 mV. The inhibition was rapid in the onset, fully reversible and did not require drug pre-perfusion. The termination of GABA and furosemide co-application was followed by transient increase in the inward current 'tail' current, which was not observed when furosemide was continuously present in the solution. The degree of furosemide block did not depend on GABA concentration. Furosemide block increased with membrane depolarization. Five millimolar furosemide depressed GABA currents by 32.4+/-1.3% at -70 mV and by 76.7+/-5.0% at +70 mV. Analysis of the voltage dependence of the block suggests that furosemide binds at the site located within GABA(A) channel pore with a dissociation constant of 5.3+/-0.5 mM at 0 mV and electric distance of 0.27. Our results provide evidence that furosemide interacts with Purkinje cell GABA(A) receptors (most probably composed of alpha1beta2/3gamma2 subunits) through a low affinity site located in channel pore and suggest that furosemide acts as a sequential open channel blocker, which prevents the dissociation of agonist while the channel is blocked.
authors.affiliation: Brain Research Institute, Russian Academy of Medical Sciences, Moscow, Russia. shar@cc.nifhi.ac.ru
authors.forename: Sergey N
authors.initials: SN
authors.lastname: Kolbaev
authors.forename: Irina N
authors.initials: IN
authors.lastname: Sharonova
authors.forename: Vladimir S
authors.initials: VS
authors.lastname: Vorobjev
authors.forename: Vladimir G
authors.initials: VG
authors.lastname: Skrebitsky
mesh_terms: D000818:Animals; D002531:Cerebellum / Q000166:cytology; D005071:Evoked Potentials / Q000187:drug effects; D005665:Furosemide / Q000494:pharmacology*; D018756:GABA Antagonists / Q000494:pharmacology*; D066298:In Vitro Techniques; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D018408:Patch-Clamp Techniques; D011689:Purkinje Cells / Q000187:drug effects / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar; D011963:Receptors, GABA-A / Q000187:drug effects / Q000502:physiology*; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology
chemical_list: D018756:GABA Antagonists; D011963:Receptors, GABA-A; D005680:gamma-Aminobutyric Acid; D005665:Furosemide