Diurnal properties of tonic and synaptic GABAA receptor-mediated currents in suprachiasmatic nucleus neurons

Abstract

Synaptic and extrasynaptic GABAA receptor (GABAAR)-mediated neurotransmission is a critical component of the suprachiasmatic nucleus (SCN) neuronal network. However, the properties of the GABAA tonic current (Itonic) and its origin remain unexplored. Spontaneous GABAA postsynaptic currents (sGPSCs) and Itonic were recorded from SCN neurons with the whole cell voltage-clamp technique at different times of the day. GABAAR antagonists (bicuculline, gabazine, and picrotoxin) inhibited sGPSC and induced an outward shift of the holding current, which defined the Itonic amplitude. The sGPSC frequency, synaptic charge transfer, and Itonic amplitude all demonstrated significant diurnal rhythms, with peaks in the middle of the day [zeitgeber time (ZT)7-8] and nadirs at night (ZT19-20). The Itonic amplitude increased proportionally with the sGPSC frequency and synaptic charge transfer during the day and required action potential-mediated GABA release, which was confirmed by TTX application. The activation of presynaptic GABAB receptors by baclofen did not significantly alter the Itonic of neurons with low-frequency sGPSC. The equilibrium potential (Eq) for Itonic was similar to the Eq for chloride and GABAA receptor-activated currents. Itonic showed outward rectification at membrane potentials over the range of -70 to -10 mV and then was linear at voltages greater than -10 mV. GABAAR containing α4-, α5-, and δ-subunits were expressed in SCN, and their contribution to Itonic was confirmed by application of the GABAAR agonist 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol (THIP) and the GABAAR inverse agonist 11,12,13,13a-tetrahydro-7-methoxy-9-oxo-9H-imidazo[1,5-a]pyrrolo[2,1-c][1,4]benzodiazepine-1-carboxylic acid ethyl ester (L655,708). Thus, the Itonic was mediated by extrasynaptic GABAARs activated predominantly by GABA diffusing out of GABAergic synapses.NEW & NOTEWORTHY A tonic current (Itonic) mediated by GABAA receptors (GABAARs) containing α4-, α5- and δ-subunits was observed in the suprachiasmatic nucleus. The Itonic amplitude strongly depended on the action potential-mediated synaptic release of GABA. The equilibrium potential for Itonic corresponds to that for GABAA currents. The frequency of GABAA postsynaptic currents and Itonic amplitude increased during the day, with peak in the middle of the day, and then gradually declined with a nadir at night, thus showing a diurnal rhythm.