Abstract
TRPV1 represents a non-selective transient receptor potential cation channel found not only in sensory neurons, but also in motor nerve endings and in skeletal muscle fibers. However, the role of TRPV1 in the functioning of the neuromuscular junction has not yet been fully established. In this study, the Levator Auris Longus muscle preparations were used to assess the effect of pharmacological activation of TRPV1 channels on neuromuscular transmission. The presence of TRPV1 channels in the nerve terminal and in the muscle fiber was confirmed by immunohistochemistry. It was verified by electrophysiology that the TRPV1 channel agonist capsaicin inhibits the acetylcholine release, and this effect was completely absent after preliminary application of the TRPV1 channel blocker SB 366791. Nerve stimulation revealed an increase of amplitude of isometric tetanic contractions upon application of capsaicin which was also eliminated after preliminary application of SB 366791. Similar data were obtained during direct muscle stimulation. Thus, pharmacological activation of TRPV1 channels affects the functioning of both the pre- and postsynaptic compartment of the neuromuscular junction. A moderate decrease in the amount of acetylcholine released from the motor nerve allows to maintain a reserve pool of the mediator to ensure a longer signal transmission process, and an increase in the force of muscle contraction, in its turn, also implies more effective physiological muscle activity in response to prolonged stimulation. This assumption is supported by the fact that when muscle was indirect stimulated with a fatigue protocol, muscle fatigue was attenuated in the presence of capsaicin.
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Acknowledgements
Authors would like to acknowledge to Assigned Spectral-Analytical Center of FRC Kazan Scientific Center of RAS for the equipment provided for the fluorescence research.
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This study was supported by the government assignment for FRC Kazan Scientific Center of RAS (122011800137-0).
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AYA and DVS: electrophysiological experiments and data analysis; NSF and AIM: tensometric recording and data analysis; LFN, ANK and MAM: immunohistochemical experiments and data analysis; AYA, DVS and AIM: conceptualization; DVS and AIM: methodology, project administration, writing, supervision.
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10571_2023_1403_MOESM1_ESM.docx
Supplementary file1 (DOCX 18 kb). SUPPLEMENTARY 1 Confocal Z-stack projection of immunolocalization of TRPV1 channels (red) in mouse m. LAL preparation. TMR-α-bungarotoxin was used as a marker of postsynaptic membrane of skeletal muscle fibers and synaptic region of neuromuscular preparation (yellow)
Supplementary file2 (AVI 34996 kb). SUPPLEMENTARY 2 Confocal Z-stack projection of immunolocalization of TRPV1 channels (red) in mouse m. LAL preparation. TMR-α-bungarotoxin was used as a marker of postsynaptic membrane of skeletal muscle fibers and synaptic region of neuromuscular preparation (yellow). Synaptophysin was used as a marker of the presynaptic membrane of the motor nerve terminal (green). The localization of each protein is presented in a separate channel. Also presented Z-stack with all channels simultaneously
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Arkhipov, A.Y., Fedorov, N.S., Nurullin, L.F. et al. Activation of TRPV1 Channels Inhibits the Release of Acetylcholine and Improves Muscle Contractility in Mice. Cell Mol Neurobiol 43, 4157–4172 (2023). https://doi.org/10.1007/s10571-023-01403-y
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DOI: https://doi.org/10.1007/s10571-023-01403-y