Functional identification of a cloned squid presynaptic voltage-dependent calcium channel

We previously cloned a voltage-dependent Ca2+ channel alpha1 subunit LoCa(v)2 cDNA from the squid optic lobe. LoCa(v)2 is designated as a non-L-type voltage-dependent Ca2+ channel based on its amino acid sequence. We performed functional expression experiments of LoCa(v)2 in oocytes and characterized the expressed currents electrophysiologically and pharmacologically. The LoCa(v)2 current was high voltage-activated and the peak current was maximal at +20 mV and lasted for long during activation. The LoCa(v)2 current was not inhibited by the drugs and toxins examined except for omega-agatoxin IVA and PLTX-II. Omega-agatoxin IVA, which is a P-type channel blocker, moderately inhibited the LoCa(v)2 current at higher concentration. PLTX-II, which blocks insect presynaptic Ca2+ channel, inhibited the LoCa(v)2 current at lower concentration. Immunohistochemical investigation showed that the LoCa(v)2 protein may exist at presynaptic terminals in the squid optic lobe. These results suggest that LoCa(v)2 is an omega-agatoxin IVA and PLTX-II-sensitive presynaptic Ca2+ channel in the squid nervous system.