J Physiol Volume 555, Number 1, 189-203, February 15, 2004 DOI: 10.1113/jphysiol.2003.055665
Activation and integration of bilateral GABA-mediated synaptic inputs in neonatal rat sympathetic preganglionic neurones in vitro
Andrew D. Whyment1,
Jennifer M. M. Wilson2,
Leo P. Renaud2 and
David Spanswick1
1 Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, UK2 Neurosciences Program, Ottawa Health Research Institute and University of Ottawa, Ottawa, Ontario, Canada, K1Y 4E9
The role of GABA receptors in synaptic transmission to neonatal rat sympathetic preganglionic neurones (SPNs) was investigated utilizing whole-cell patch clamp recording techniques in longitudinal and transverse spinal cord slice preparations. In the presence of glutamate receptor antagonists (NBQX, 5 µM and D-APV, 10 µM), electrical stimulation of the ipsilateral or contralateral lateral funiculi (iLF and cLF, respectively) revealed monosynaptic inhibitory postsynaptic potentials (IPSPs) in 75% and 65% of SPNs, respectively. IPSPs were sensitive to bicuculline (10 µM) in all neurones tested and reversed polarity around -55 mV, the latter indicating mediation via chloride conductances. In three neurones IPSPs evoked by stimulation of the iLF (n= 1) or cLF (n= 2) were partly sensitive to strychnine (2 µM). The expression of postsynaptic GABAA and GABAB receptors were confirmed by the sensitivity of SPNs to agonists, GABA (2 mM), muscimol (10100 µM) or baclofen (10100 µM), in the presence of TTX, each of which produced membrane hyperpolarization in all SPNs tested. Muscimol-induced responses were sensitive to bicuculline (110 µM) and SR95531 (10 µM) and baclofen-induced responses were sensitive to 2-hydroxy-saclofen (100200 µM) and CGP55845(200 nM). The GABAC receptor agonist CACA (200 µM) was without significant effect on SPNs. These results suggest that SPNs possess postsynaptic GABAA and GABAB receptors and that subsets of SPNs receive bilateral GABAergic inputs which activate GABAA receptors, coupled to a chloride conductance. At resting or holding potentials close to threshold either single or bursts (10100 Hz) of IPSPs gave rise to a rebound excitation and action potential firing at the termination of the burst. This effect was mimicked by injection of small (1020 pA) rectangular-wave current pulses, which revealed a time-dependent, Cs+-sensitive inward rectification and rebound excitation at the termination of the response to current injection. Synaptic activation of a rebound excitation mediated by a time-dependent inward rectification expressed intrinsically by SPNs may provide a novel mechanism enabling SPNs to be entrained to rhythms driven from the brainstem or higher centres.
(Received 23 September 2003;
accepted after revision 9 December 2003;
first published online 12 December 2003)
Corresponding author D. Spanswick: Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, UK. Email: d.c.spanswick{at}warwick.ac.uk
Copyright © 2004 The Physiological Society.