Increases in intracellular calcium triggered by channelrhodopsin-2 potentiate the response of metabotropic glutamate receptor mGluR7

The metabotropic glutamate receptor 7a (mGluR7a), a hepta-helical G alpha(i/o)-coupled protein, has been shown to be important for presynaptic feedback inhibition at central synapses and certain forms of long term potentiation and long term depression. The intracellular C terminus of mGluR7a interacts with calmodulin in a Ca2+-dependent manner, and calmodulin antagonists have been found to abolish presynaptic inhibition of glutamate release in neurons and mGluR7a-induced activation of G-protein-activated inwardly rectifying K+ channel (GIRK) channels in HEK293 cells. Here, we characterized the Ca2+ dependence of mGluR7a signaling in Xenopus oocytes by using channelrhodopsin-2 (ChR2), a Ca2+-permeable, light-activated ion channel for triggering Ca2+ influx, and a GIRK3.1/3.2 concatemer to monitor mGluR7a responses. Application of the agonist (S)-2-amino- 4-phosphonobutanoic acid (L-AP4) (1-100 mu M) caused a dose-dependent inward current in high K+ solutions due to activation of GIRK channels by G-protein beta gamma subunits released from mGluR7a. Elevation of intracellular free Ca2+ by light stimulation of ChR2 markedly increased the amplitude of L-AP4 responses, and this effect was attenuated by the calcium chelator 1,2-bis(2-aminophenoxy) ethane-N,N,N',N'-tetraacetic acid tetrakis (acetoxymethyl ester). L-AP4 responses were potentiated by submembranous [Ca2+] levels within physiological ranges and with a threshold close to resting [Ca2+](i) values, as determined by recording the endogenous Xenopus Ca2+-activated chloride conductance. Together, these results show that L-AP4-dependent mGluR7a signaling is potentiated by physiological levels of [Ca2+](i), consistent with a model in which presynaptic mGluR7a acts as a coincidence detector of Ca2+ influx and glutamate release.