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The Journal of Neuroscience, January 15, 1999, 19(2):737-746

ATP Stimulates Sympathetic Transmitter Release via Presynaptic P2X Purinoceptors

Stefan Boehm

Department of Neuropharmacology, University of Vienna, A-1090 Vienna, Austria

ATP is a fast transmitter in sympathetic ganglia and at the sympathoeffector junction. In primary cultures of dissociated rat superior cervical ganglion neurons, ATP elicits noradrenaline release in an entirely Ca²⁺-dependent manner. Nevertheless, ATP-evoked noradrenaline release was only partially reduced (by ~50%) when either Na⁺ or Ca²⁺ channels were blocked, which indicates that ATP receptors themselves mediated transmembrane Ca²⁺ entry. An "axonal" preparation was obtained by removing ganglia from explant cultures, which left a network of neurites behind; immunostaining for axonal and dendritic markers revealed that all of these neurites were axons. In this preparation, ATP raised intraaxonal Ca²⁺ and triggered noradrenaline release, and these actions were not altered when Ca²⁺ channels were blocked by Cd²⁺. Hence, Ca²⁺-permeable ATP-gated ion channels, i.e., P2X purinoceptors, are located at presynaptic sites and directly mediate Ca²⁺-dependent transmitter release. These presynaptic P2X receptors displayed a rank order of agonist potency of ATP  2-methylthio-ATP > ATPS ,-methylene-ATP  ,-methylene-L-ATP and were blocked by suramin or PPADS. ATP, 2-methylthio-ATP, and ATPS also evoked inward currents measured at neuronal somata, but there these agonists were equipotent. Hence, presynaptic P2X receptors resemble the cloned P2X2 subtype, but they appear to differ from somatodendritic P2X receptors in terms of agonist sensitivity. Suramin reduced depolarization-evoked noradrenaline release by up to 20%, when autoinhibitory mechanisms were inactivated by pertussis toxin. These results indicate that presynaptic P2X purinoceptors mediate a positive, whereas G-protein-coupled P2Y purinoceptors mediate a negative, feedback modulation of sympathetic transmitter release.

Key words: rat superior cervical ganglion neurons; noradrenaline release; ATP; P2X receptors; presynaptic modulation; intracellular Ca²⁺

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Copyright © 1999 Society for Neuroscience  0270-6474/99/192737-10$05.00/0

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