Protein kinase C-mediated inhibition of recombinant T-type Cav3.2 channels by neurokinin 1 receptors.
The voltage-activated T-type calcium channel (Ca(V)3.2) and the G protein-coupled neurokinin 1 (NK1) receptor are expressed in peripheral tissues and in central neurons, in which they participate in diverse physiological processes, including neurogenic inflammation and nociception. In the present report, we demonstrate that recombinant Ca(V)3.2 channels are reversibly inhibited by NK1 receptors when both proteins are transiently coexpressed in human embryonic kidney 293 cells. We found that the voltage-dependent macroscopic properties of Ca(V)3.2 currents were unaffected during NK1 receptor-mediated inhibition. However, inhibition was attenuated in cells coexpressing either the dominant-negative Galpha(q) Q209L/D277N or the regulator of G protein signaling (RGS) proteins 2 (RGS2) and 3T (RGS3T), which are effective antagonists of Galpha(q/11). By contrast, inhibition was unaffected in cells coexpressing human rod transducin (Galpha(t)), which buffers Gbetagamma. Channel inhibition was blocked by 1-[6-[[17beta-methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-1H-pyrrole-2,5-dione (U73122) and bisindolylmaleimide I, selective inhibitors of phospholipase Cbeta and protein kinase C (PKC), respectively. Inhibition was occluded by application of the PKC activator phorbol-12-myristate-13-acetate. Altogether, these data indicate that NK1 receptors inhibit Ca(V)3.2 channels through a voltage-independent signaling pathway that involves Galpha(q/11), phospholipase Cbeta, and PKC. Our results provide novel evidence regarding the mechanisms underlying T-type calcium channel modulation by G protein-coupled receptors. Functional coupling between Ca(V)3.2 channels and NK1 receptors may be relevant in neurogenic inflammation, neuronal rhythmogenesis, nociception, and other physiological processes.
Rangel
Azahel
A
Departamento de Fisiología, Universidad Autónoma de San Luis Potosí, Av. Venustiano Carranza, San Luis Potosí, México.
Sánchez-Armass
Sergio
S
Meza
Ulises
U
D020747:Calcium Channels, T-Type / Q000378:metabolism* / Q000502:physiology; D002460:Cell Line; D006801:Humans; D011493:Protein Kinase C / Q000502:physiology*; D018040:Receptors, Neurokinin-1 / Q000502:physiology*; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D015398:Signal Transduction / Q000502:physiology
C479952:CACNA1H protein, human; D020747:Calcium Channels, T-Type; D018040:Receptors, Neurokinin-1; D011994:Recombinant Proteins; D011493:Protein Kinase C