Potentiation by ATP of the Postsynaptic Acetylcholine Response at Developing Neuromuscular Synapses in Xenopus Cell Cultures

1. Extracellular application of ATP to developing Xenopus neuromuscular synapses in culture resulted in a marked increase in the amplitude and frequency of spontaneous synaptic currents, using whole-cell recording. 2. The postsynaptic action of ATP was examined by studying the response of isolated muscle cells to iontophoretically applied acetylcholine (ACh). ATP enhanced the responses of the muscle membrane to ACh. The order of potency for various nucleotides (ATP = ADP >> AMP, adenosine, GTP) suggests that ATP acts through P2-purinoceptors. The effect of ATP on whole-cell currents was also abolished by the protein kinase inhibitor H-7. 3. Single-channel measurements indicate that ATP increased the mean open time of low-conductance ACh channels. No change in the conductance of ACh channels was observed. 4. Local application of ATP to one region of the elongated myocyte surface resulted in potentiated ACh responses only at the ATP-treated region, suggesting that the cytosolic second messengers were effectively confined within the muscle cytoplasm. 5. The results of the present study suggest that ATP released from the nerve terminals may potentiate the ACh response of developing muscle cells during the early phase of synaptogenesis, and that the action of ATP can be restricted to the subsynaptic region exposed to the secreted ATP.