Activation of D2-like dopamine receptors inhibits GABA and glycinergic neurotransmission to pre-motor cardiac vagal neurons in the nucleus ambiguus

Abstract

The parasympathetic control of heart rate arises from premotor cardiac vagal neurons (CVNs) located in the nucleus ambiguus (NA). Previous microinjection studies in NA show that dopamine evokes a decrease in heart rate, but the underlying mechanisms responsible for these responses were not identified. This study tested whether dopamine modulates inhibitory GABAergic and glycinergic and/or excitatory glutamatergic neurotransmission to CVNs. Retrogradely labeled CVNs were identified in an in vitro rat brainstem slice preparation and synaptic events were recorded using whole cell voltage clamp techniques. Bath application of dopamine (100 μM) had no effect on excitatory synaptic events, but reversibly inhibited the frequency (but not amplitude) of GABAergic inhibitory postsynaptic currents (IPSCs) in CVNs. Similarly, dopamine (10 μM and 100 μM) inhibited glycinergic IPSC frequency by ~50% and 70% respectively. The reduction in inhibitory neurotransmission to CVNs by dopamine was prevented by the sodium channel blocker TTX (1μM) indicating that the dopamine mediated effects were action potential dependent. Dopamine evoked responses were mimicked by the D2-like receptor agonist, Quinpirole but not D1-like receptor agonist, SKF 38393. In addition, the dopamine mediated depression of inhibitory synaptic responses were prevented by the D2-like receptor antagonist sulpiride, but not by D1-like or adrenergic or serotonergic receptor antagonists, suggesting that these responses were D2-like receptor mediated and not D1-like or adrenergic or 5-HT receptor mediated. These data suggest that dopamine acts via dis-inhibition, and diminishes inhibitory GABAergic and glycinergic neurotransmission to CVNs, which would be predicted to increase parasympathetic activity to the heart and evoke a bradycardia.

Keywords: 2-amino-5-phosphonopentanoic acid; 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; 5-HT; 6-cyano-7-nitroquinoxaline; CNQX; CVN; DL-AP(5); DMNX; EGTA; EPSC; HEPES; IPSC; LC; N-methyl-d-aspartic acid; NA; NE; NMDA; NTS; PSS; RVLM; TTX; Tetrodotoxin; cardiac vagal neuron; dopamine; dorsal motor nucleus of the vagus; ethylene glycol tetraacetic acid; excitatory postsynaptic current; heart rate; inhibitory postsynaptic current; locus coerulus; mIPSC; miniature inhibitory postsynaptic current; norepinephrine; nucleus ambiguus; nucleus tractus solitarius; parasympathetic; physiological salt solution; rostroventrolateral medulla; serotonin.