Kinetic and pharmacological properties of GABA(A) receptors in single thalamic neurons and GABA(A) subunit expression

Synaptic inhibition in the thalamus plays critical roles in sensory process ing and thalamocortical rhythm generation. To determine kinetic, pharmacolo gical, and structural properties of thalamic gamma -aminobutyric acid type A (GABA(A)) receptors, we used patch-clamp techniques and single-cell rever se transcriptase polymerase chain reaction (RT-PCR) in neurons from two pri ncipal rat thalamic nuclei-the reticular nucleus (nRt) and the ventrobasal (VB) complex. Single-channel recordings identified GABA(A) channels with de nsities threefold higher in VB than nRt neurons, and with mean open time fo urfold longer for nRt than VB [14.6 +/-2.5 vs. 3.8 +/-0.7 (SE) ms, respecti vely]. GABA(A) receptors in nRt and VB cells were pharmacologically distinc t. Zn2+ (100 muM) reduced GABA(A) channel activity in VB and nRt by 84 and 24%, respectively. Clonazepam (100 nM) increased inhibitory postsynaptic cu rrent (IPSC) decay time constants in nRt (from 44.3 to 77.9 ms, P<0.01) but not in VB. Single-cell RT-PCR revealed subunit heterogeneity between nRt a nd VB cells. VB neurons expressed <alpha>1-alpha3, alpha5, beta1-3, gamma2- 3, and delta, while nRt cells expressed alpha3, alpha5, gamma2- 3, and delt a. Both cell types expressed more subunits than needed for a single recepto r type, suggesting the possibility of GABA(A) receptor heterogeneity within individual thalamic neurons. beta -subunits were not detected in nRt cells , which is consistent with very low levels reported in previous in situ hyb ridization studies but inconsistent with the expected dependence of functio nal GABA(A) receptors on beta subunits. Different single-channel open times likely underlie distinct IPSC decay time constants in VB and nRt cells. Wh ile we can make no conclusion regarding b subunits, our findings do support a subunits, possibly alpha1 versus alpha3, as structural determinants of c hannel deactivation kinetics and clonazepam sensitivity. As the gamma2 and delta subunits previously implicated in Zn2+ sensitivity are both expressed in each cell type, the observed differential Zn2+ actions at VB versus nRt GABA(A) receptors may involve other subunit differences.