Channel properties determine the transient activation kinetics of recombinant GABA(A) receptors.
To understand the mechanisms underlying activation and deactivation of GABA(A) receptor (GABAR) channels, we studied the properties of an identified GABAR isoform under conditions similar to those present at central GABAergic synapses. Recombinant alpha5beta3gamma2L GABARs were expressed in L929 fibroblasts and studied using patch-clamp recording techniques. Brief application of a high GABA concentration to outside-out membrane patches elicited transient currents that resembled those reported for miniature inhibitory postsynaptic currents (mIPSCs), as well as native GABAR currents recorded under similar conditions. Characteristic of these currents was a rapid activation phase followed by a prolonged biphasic deactivation phase that far outlasted GABA application. Single-channel recordings revealed unique patterns of channel activity with two channel conductance states of 22 and 16 pS. The prolonged deactivation phase appeared to be sustained by entry into and reopening from long-lasting closures or desensitized states. Agonist affinity determined the time course of deactivation, indicating that occupied receptors drove the channel activity underlying the decay of current. The time course of deactivation was also longer at depolarized membrane potentials. The similarities between transient activation kinetics of recombinant alpha5beta3gamma2L GABARs to activation of synaptic GABARs (rapid activation and prolonged, voltage-dependent deactivation) suggest that intrinsic channel properties determine much of the response patterns of native GABARs.
Burgard
E C
EC
Department of Neurology, University of Michigan, Ann Arbor, MI 48104-1687, USA.
Haas
K F
KF
Macdonald
R L
RL
D000818:Animals; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D018755:GABA Agonists / Q000494:pharmacology; D007555:Isoxazoles / Q000494:pharmacology; D007700:Kinetics; D008564:Membrane Potentials / Q000187:drug effects; D020033:Protein Isoforms / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D011963:Receptors, GABA-A / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D013997:Time Factors; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology*
D018755:GABA Agonists; D007555:Isoxazoles; D020033:Protein Isoforms; D011963:Receptors, GABA-A; D011993:Recombinant Fusion Proteins; D005680:gamma-Aminobutyric Acid; C015542:gaboxadol