The ionic basis of adenosine receptor actions on post-ganglionic neurones in the rat.

Adenosine inhibited three Ca2+-dependent potentials recorded intracellularly from post-ganglionic neurones of the rat superior cervical ganglion. A shoulder on the falling phase of the action potential elicited in normal Locke solution, a hyperpolarizing after-potential (h.a.p.) that follows the spike, and a regenerative Ca2+ spike elicited in Locke solution containing TTX and TEA were all reversibly inhibited by adenosine analogues in a dose-dependent fashion. The maximum rate of rise of the Ca2+ spike (dV/dt) was markedly reduced suggesting that the underlying mechanism of adenosine action is inhibition of the Ca2+ conductance mechanism and thus, the voltage-sensitive Ca2+ current. I/V curves in low Ca2+, high Mg2+, TTX, TEA, and Co2+ to block the Ca2+ current show no change in resistance in the presence of 2-chloroadenosine. The actions of adenosine were nearly eliminated in the presence of 1 mM-theophylline, an adenosine receptor antagonist. The order of agonist potency on the inhibition of the h.a.p. was: N-6-[L-phenylisopropyl] adenosine (L-PIA) greater than 2-chloroadenosine greater than adenosine greater than cyclic AMP = 5' AMP. The concentration of L-PIA which produced a half-maximal effect (EC50) was 0.5 microM and that for cyclic AMP was 100 microM. Dipyridamole, an adenosine uptake blocker, potentiated the effects of low concentrations of adenosine and shifted the dose-response curve for adenosine towards that of 2-chloroadenosine (EC50 = 1 microM). These results are consistent with the concept of an external adenosine receptor, but we are unable to assign a receptor subtype. Cyclic AMP mimicked the effects of adenosine, but these effects were eliminated by adenosine deaminase. Our results suggest that the electrogenic effects of bath-applied cyclic AMP may result from the metabolism of cyclic AMP to adenosine by ganglionic tissue. We conclude that adenosine activates a receptor on the neuronal cell surface to inhibit the voltage-dependent Ca2+ current.

Calcium currents modulated by adrenergic receptors in sympathetic neurons.

The superior cervical sympathetic ganglion is currently being used as a model neuronal system for the study of Ca2+-dependent processes in the mammalian nervous system. We have characterized a regenerative calcium conductance in postganglionic neurons. This Ca2+ current contributes to the shoulder of the action potential. In addition, Ca2+ influx during the spike activates a K+ conductance, which generates a hyperpolarizing afterpotential. These Ca2+-dependent potentials are antagonized by catecholamines. Pharmacologic studies suggest that alpha 2-adrenergic receptors inhibit the regenerative voltage-dependent Ca2+ influx that occurs during the action potential. Alpha-adrenergic agonists were also found to reduce the depression of the compound action potential following a train of preganglionic stimuli. We hypothesize that alpha 2-receptors function primarily to antagonize Ca2+ influx and thereby exert significant control over neuronal excitability and release of neurotransmitters.

The action of cAMP and catecholamines in mammalian sympathetic ganglia.

Electrophysiological approaches using intracellular microelectrode techniques have failed to critically test the hypothesis that cyclic AMP (cAMP) mediates the slow inhibitory postsynpatic potential (IPSP). The slow IPSP is not readily elicited, and the resting membrane potential is relatively insensitive to application of catecholamines and adenine nucleotides. However, comprehensive studies of voltage-dependent events in postganglionic neurons reveal three Ca2+-dependent potentials that are quite sensitive to catecholamines and adenine nucleotides. The hyperpolarizing afterpotential, the action potential shoulder, and the Ca2+ spike are all inhibited by alpha-adrenergic agonists, adenosine, and cAMP. We have proposed that simulation of alpha-adrenergic and adenosine receptors on the post-synaptic membrane results in antagonism of an inward Ca2+ current. Further experimentation is necessary to determine if cAMO acts as a second messenger or only by activating an adenosine receptor. Preliminary studies suggest that catecholamines and adenine nucleotides have similar and potent actions on the terminals of preganglionic axons. Here, inhibition of Ca2+ influx results in reduced acetylcholine release but facilitates high-frequency cholinergic transmission. More quantitative biophysical and pharmacological studies are required to better characterize the synaptic mechanisms in sympathetic ganglia.