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STUDIES ON THE SYNAPTIC FACILITATION EFFECT OF HYDROGEN PEROXIDE IN THE DEVELOPING NEUROMUSCULAR SYNAPSE

We have previously demonstrated that muscle‐derived IGF‐1 is important in the development of neuromuscular synapse and hydrogen peroxide (H2O2) plays an important role in the release of IGF‐1. Here we further test the role of H2O2 in the development of neuromuscular synapse of Xenopus laevis by using the recording of whole‐cell patch clamp. Bath application of H2O2 dose‐dependently potentiates the frequency of spontaneous ACh release in Day‐1 Xenopus neuromuscular junction. The dose‐response curve shift to left while the facilitation effect of H2O2 on SSC frequency was test in Day‐3 cultures, suggesting the sensitivity of H2O2‐induced facilitating effect reduced as synapse matured. Pretreatment of the culture with H2O2 scavengers both N‐Acetylcysteine or sodium pyruvate for 1 day significantly hampered the development of synapse as the SSC frequency and amplitude both significant reduced in treated Day‐1 synapses. We next test the effect of H2O2 on Day‐1 N‐Acetylcysteine/sodium pyruvate‐pretreated synapse was test after extensively washed the culture to removed N‐Acetylcysteine/sodium pyruvate. The SSC frequency facilitating effect induced by H2O2 was significantly enhanced in treated synapse, which supports the notion that H2O2 in involved in the development of synapse. Furthermore, co‐pretreatment the culture with insulin in N‐Acetylcysteine/sodium pyruvate group reversed N‐Acetylcysteine/sodium pyruvate treatment‐induced synapse immaturity. Overall, results from our current studies provide strong supports on the involvement of H2O2 in the early development of neuromuscular synapse.