ATP acts as fast neurotransmitter in rat habenula: neurochemical and enzymecytochemical evidence.

The release of ATP and ADP, the putative central neurotransmitters, from the isolated habenula preparation was investigated in the rat, at rest and during electrical stimulation, using the luciferin-luciferase assay and the creatine phosphokinase assay. Electrical field stimulation (2 Hz, 360 pulses) released a considerable amount of ATP (2450 +/- 280 pmol/g wet tissue) from the tissue; inhibition of the voltage Na+ entry by tetrodotoxin (1 microM) reduced significantly the evoked release (by 66.25 +/- 6.65%), but not the resting release of ATP. Endogenous ADP also appeared in the effluent, but its amount differed during resting condition and after stimulation from that of ATP, suggesting that the majority of the released compound is ATP in response to stimulation. When ATP was added to the tissue, it readily decomposed to ADP and AMP (Km = 811.6 +/- 68.88 microM, vmax = 23.1 +/- 2.75 nmol/min per prep., measured by high-performance liquid chromatography combined with ultraviolet detection), indicating that the habenula contains ectoATPases. In addition, the inactivation of extracellular ATP by the ectoATPase enzyme was also visualized by electron microscopic enzyme cytochemistry. The ectoATPase enzyme was present on the membranes of the dendrites and nerve terminals and in the synapses of the habenula. Taking into account the fact that ATP is ubiquitous in excitable cells (storage) and the findings published by Edwards et al. in 1992 ("ATP receptor-mediated synaptic currents in the central nervous system", Nature, Vol. 359, pp. 144-147), our data provides evidence for the release by axonal stimulation and extracellular decomposition of ATP, all needed for an endogenous substance qualified as a transmitter.