A Store-Operated Ca2+ Influx Pathway in the Bag Cell Neurons of Aplysia

Although store-operated Ca2+ influx has been well-studied in nonneuronal cells, an understanding of its nature in neurons remains poor. In the bag cell neurons of Aplysia californica , prior work has suggested that a Ca2+ entry pathway can be activated by Ca2+ store depletion. Using fura-based imaging of intracellular Ca2+ in cultured bag cell neurons, we now characterize this pathway as store-operated Ca2+ influx. In the absence of extracellular Ca2+, the endoplasmic reticulum Ca2+-ATPase inhibitors, cyclopiazonic acid (CPA) or thapsigargin, depleted intracellular stores and elevated intracellular free Ca2+. With the subsequent addition of extracellular Ca2+, a prominent Ca2+ influx was observed. The ryanodine receptor agonist, chloroethylphenol (CEP), also increased intracellular Ca2+ but did not initiate store-operated Ca2+ influx, despite overlap between CEP- and CPA-sensitive stores. Bafilomycin A, a vesicular H+-ATPase inhibitor, liberated intracellular Ca2+ from acidic stores and attenuated subsequent Ca2+ influx, presumably by replenishing CPA-depleted stores. Store-operated Ca2+ influx was partially blocked by low concentrations of La3+ or BTP2, and strongly inhibited by either 1-[b-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl]-1H-imidazole (SKF-96365) or a high concentration of Ni2+. Regarding IP3 receptor blockers, 2-aminoethyldiphenyl borate, but not xestospongin C, prevented store-operated Ca2+ influx. However, jasplakinolide, an actin stabilizer reported to inhibit this pathway in smooth muscle cell lines, was ineffective. The bag cell neurons initiate reproductive behavior through a prolonged afterdischarge associated with intracellular Ca2+ release and neuropeptide secretion. Store-operated Ca2+ influx may serve to replenish stores depleted during the afterdischarge or participate in the release of peptide that triggers behavior.