Orexin-A potentiates glycine currents by activating OX1R and IP3/Ca2+/PKC signaling pathways in spinal cord ventral horn neurons

Abstract

Orexin-A/B modulates multiple physical functions by activating their receptors (OX₁R and OX₂R), but its effects in the spinal cord motor control remain unknown. Using acute separation (by digestive enzyme) of cells and patch-clamp recordings, we aimed to investigate the effect and mechanisms of orexin-A on the glycine receptors in the spinal cord ventral horn neurons. Orexin-A potentiated the glycine currents by activating OX₁R. In Ca²⁺-free extracellular solution, orexin-A still increased the glycine currents. While, the orexin-A-induced potentiation was blocked when Ca²⁺ was chelated by internal infusion of BAPTA, and the orexin-A effect was abolished by the IP₃ receptor antagonists heparin and Xe-C. The PKC inhibitor Bis-IV nullified the orexin-A effect. In addition, orexin-A did not cause a further enhancement of the glycine currents after bath application of the PKC activator PMA. In conclusion, after OX₁R is activated, a distinct IP₃/Ca²⁺-dependent PKC signaling pathway, is likely responsible for the orexin-A potentiation on glycine currents in the spinal cord ventral horn neurons.

Keywords: Glycine receptors (GlyRs); Orexin receptors (OXRs); Orexin-A; Patch-clamp recordings; Spinal cord ventral horn neurons.