ASIC3 knockout alters expression and activity of P2X3 in muscle afferent nerves of rat model of peripheral artery disease

Abstract

Abstract In peripheral artery disease (PAD), the metaboreceptor and mechanoreceptor in muscle afferent nerves contribute to accentuated sympathetic outflow via a neural reflex termed exercise pressor reflex (EPR). Particularly, lactic acid and adenosine triphosphate (ATP) produced in exercising muscles respectively stimulate acid sensing ion channel subtype 3 (ASIC3) and P2X3 receptors (P2X3) in muscle afferent nerves, inducing the reflex sympathetic and BP responses. Previous studies indicated that those two receptors are spatially close to each other and AISC3 may have a regulatory effect on the function of P2X3. This inspired our investigation on the P2X3‐mediated EPR response following AISC3 abolished, which was anticipated to shed light on the future pharmacological and genetic treatment strategy for PAD. Thus, we tested the experimental hypothesis that the pressor response to P2X3 stimulation is greater in PAD rats with 3 days of femoral artery occlusion and the sensitizing effects of P2X3 are attenuated following ASIC3 knockout (KO) in PAD. Our data demonstrated that in wild type (WT) rats femoral occlusion exaggerated BP response to activation of P2X3 using α,β‐methylene ATP injected into the arterial blood supply of the hindlimb, meanwhile the western blot analysis suggested upregulation of P2X3 expression in dorsal root ganglion supplying the afferent nerves. Using the whole cell patch‐clamp method, we also showed that P2X3 stimulation enhanced the amplitude of induced currents in muscle afferent neurons of PAD rats. Of note, amplification of the P2X3 evoked‐pressor response and expression and current response of P2X3 was attenuated in ASIC3 KO rats. We concluded that the exaggerated P2X3‐mediated pressor response in PAD rats is blunted by ASIC3 KO due to the decreased expression and activities of P2X3 in muscle afferent neurons.

Keywords

• ASIC
• blood pressure
• dorsal root ganglion
• hindlimb ischemia
• muscle afferent
• P2X