Anda di halaman 1dari 2

Effect of Substance P on the release of acetylcholine at the neuromuscular junction.

Rafaela Pinto Coelho Santos, Thalita Mota Caldeira, Christopher Kushmerick, Ligia Araujo Naves Dept. de Fisiologia e Biofsca Instituto de Cincias Biolgicas Universidade Federal de Minas Gerais, Belo Horizonte- MG, Brasil Introduction and Objectives: Quantal content is a measure of the quantity of neurotransmitter released in response to nerve stimulation. Many nerve terminals express receptors for neurotransmitters or hormones that modulate transmitter release. At the neuromuscular junction, receptors for angiotensins, cannabinoids and vanilloides have been shown to modulate quantal content (Bevilaqua et al., 2002; Oliveira et al., 2007; Silveira et al., 2010). Substance P is a neuropeptide produced in the cells bodies of certain neuron and released from nerve terminal during inflamation and pain processes. Previous studies indicate that Substance P increases the excitability of cholinergic motoneuron cell bodies (Adachi et al., 2010) and increases quantal content at the neuromuscular junction (Akasu, 1986). Our long-term goal is to determine the receptor type and mechanism by which Substance P affects transmitter release from motoneuron nerve terminals. As a first step, we present results confirming that Subtance P increases quantal content at the frog neuromuscular junction. In addition, we also provide evidence that Substance P increases the threshold of the motoneuron. Methods: Cutaneus pectoris muscles and attached nerves were isolated from young adult Rana catesbeiana in normal frog Ringer. The nerve was stimulated via a suction electrode with pulses of 0.08 ms duration and threshold was measured as the smallest voltage able to produce a muscle contraction. After measuring threshold, the bathing solution was exchanged for low-Ca2+ / high Mg2+ Ringer to reduce the size of endplate potencials (EPPs) and thus block muscle contraction. The muscle was impaled using standard intracellular techniques and correct placement of the electrode near the endplate was identified by the presence of miniature EPPs. The nerve was stimulated using paired pulses (15 ms interval) at three times its threshold and the fraction of stimuli not followed by an EPP was used to estimate the average quantal content of the first and second EPP using the method of failures. Facilitation was measured as the ratio of quantal content of the second EPP to the first.

Results and Conclusion: Substancia P 1M increased quantal content of evoked EPPs by 38 27% (mean 95%confidence limits), N=12. However the 3M there was no significant effect. There was no significant effect on facilitation. We conclude that substance P acts on the motoneuron terminal to increase transmitter release.