Appearance of a novel Ca2+ influx pathway in Sf9 insect cells following expression of the transient receptor potential-like (trpl) protein of Drosophila

Abstract

Activation of phospholipase C, elevation of free cytosolic Ca2+ concentration ([Ca2+]i) and stimulation of Ca2+ influx have been implicated in Drosophila phototransduction. Electrophysiological studies suggest that trp and trpl proteins may be important for the light-activated Ca2+ current found in Drosophila photoreceptor cells. Although these proteins exhibit homologies to voltage-gated Ca2+ and Na+ channels, their actual function in insect cells and their relation to proteins involved in mammalian cell Ca2+ signaling remains unknown. In the present study, [Ca2+]i was examined in fura-2-loaded Sf9 insect cells infected with recombinant baculovirus containing cDNA for the trpl protein. Ca2+ influx was examined by use of Ba2+, a Ca2+ surrogate that is not a substrate for Ca(2+)-pumps or carriers and by measurement of whole-cell membrane currents. The results suggest that expression of trpl is associated with appearance of a Ca2+ permeable, non-selective cation channel formed by the trpl protein.