Transmitter modulation of spike-evoked calcium transients in arousal related neurons: muscarinic inhibition of SNX-482-sensitive calcium influx

Nitric oxide synthase (NOS)-containing cholinergic neurons in the laterodorsal tegmentum (LDT) influence behavioral and motivational states through their projections to the thalamus, ventral tegmental area and a brainstem 'rapid eye movement (REM)-induction' site. Action potential-evoked intracellular calcium transients dampen excitability and stimulate NO production in these neurons. In this study, we investigated the action of several arousal-related neurotransmitters and the role of specific calcium channels in these LDT Ca2+-transients by simultaneous whole-cell recording and calcium imaging in mouse (P14-P30) brain slices. Carbachol, noradrenaline and adenosine inhibited spike-evoked Ca2+-transients, while histamine, t-ACPD, a metabotropic glutamate receptor agonist, and orexin-A did not. Carbachol inhibition was blocked by atropine, was insensitive to blockade of G-protein-coupled inward rectifier (GIRK) channels and was not inhibited by nifedipine, omega-conotoxin GVIA or omega-agatoxin IVA, which block L-, N- and P/Q-type calcium channels, respectively. In contrast, SNX-482 (100 nm), a selective antagonist of R-type calcium channels containing the alpha1E (Cav2.3) subunit, attenuated carbachol inhibition of the somatic spike-evoked calcium transient. To our knowledge, this is the first demonstration of muscarinic inhibition of native SNX-482-sensitive R-channels. Our findings indicate that muscarinic modulation of these channels plays an important role in the feedback control of cholinergic LDT neurons and that inhibition of spike-evoked Ca2+-transients is a common action of neurotransmitters that also activate GIRK channels in these neurons. Because spike-evoked calcium influx dampens excitability, our findings suggest that these 'inhibitory' transmitters could boost firing rate and enhance responsiveness to excitatory inputs during states of high firing, such as waking and REM sleep.