L-cysteine modulates visceral nociception mediated by the Ca(V)2.3 R-type calcium channels

Ca(V)2.3 channels are subthreshold voltage-gated calcium channels that play crucial roles in neurotransmitter release and regulation of membrane excitability, yet modulation of these channels with endogenous molecules and their role in pain processing is not well studied. Here, we hypothesized that an endogenous amino acid l-cysteine could be a modulator of these channels and may affect pain processing in mice. To test this hypothesis, we employed conventional patch-clamp technique in the whole-cell configuration using recombinant Ca(V)2.3 subunit stably expressed in human embryonic kidney (HEK-293) cells. We found in our in vitro experiments that l-cysteine facilitated gating and increased the amplitudes of recombinant Ca(V)2.3 currents likely by chelating trace metals that tonically inhibit the channel. In addition, we took advantage of mouse genetics in vivo using the acetic acid visceral pain model that was performed on wildtype and homozygous Cacna1e knockout male littermates. In ensuing in vivo experiments, we found that l-cysteine administered both subcutaneously and intraperitoneally evoked more prominent pain responses in the wildtype mice, while the effect was completely abolished in knockout mice. Conversely, intrathecal administration of l-cysteine lowered visceral pain response in the wildtype mice, and again the effect was completely abolished in the knockout mice. Our study strongly suggests that l-cysteine-mediated modulation of Ca(V)2.3 channels plays an important role in visceral pain processing. Furthermore, our data are consistent with the contrasting roles of Ca(V)2.3 channels in mediating visceral nociception in the peripheral and central pain pathways.

Automated summary

This study demonstrates that the endogenous amino acid l-cysteine can modulate Ca(V)2.3 channels and plays an important role in visceral pain processing. The results also reveal contrasting roles of Ca(V)2.3 channels in mediating visceral nociception in the peripheral and central pain pathways.