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The Journal of Neuroscience, August 1, 2000, 20(15):5748-5755
Metabolic Stress Reversibly Activates the Drosophila Light-Sensitive Channels TRP and TRPL In Vivo
Keren Agam1, Mark von Campenhausen2, Simon Levy3, Hagit Cohen Ben-Ami1, Boaz Cook1, Kuno Kirschfeld2, and Baruch Minke1 1 Department of Physiology and the Kühne Minerva Center for Studies of Visual Transduction, The Hebrew University-Hadassah Medical School, Jerusalem 91120, Israel, 2 Max Planck-Institut für biologische Kybernetik, 72076, Tübingen, Germany, and 3 Department of Physiology, Boston University School of Medicine, Boston, Massachusetts 02118
Drosophila transient receptor potential (TRP) is a prototypical member of a novel family of channel proteins underlying phosphoinositide-mediated Ca2+ entry. Although the initial stages of this signaling cascade are well known, downstream events leading to the opening of the TRP channels are still obscure. In the present study we applied patch-clamp whole-cell recordings and measurements of Ca2+ concentration by ion-selective microelectrodes in eyes of normal and mutant Drosophila to isolate the TRP and TRP-like (TRPL)-dependent currents. We report that anoxia rapidly and reversibly depolarizes the photoreceptors and induces Ca2+ influx into these cells in the dark. We further show that openings of the light-sensitive channels, which mediate these effects, can be obtained by mitochondrial uncouplers or by depletion of ATP in photoreceptor cells, whereas the effects of illumination and all forms of metabolic stress were additive. Effects similar to those found in wild-type flies were also found in mutants with strong defects in rhodopsin, Gq-protein, or phospholipase C, thus indicating that the metabolic stress operates at a late stage of the phototransduction cascade. Genetic elimination of both TRP and TRPL channels prevented the effects of anoxia, mitochondrial uncouplers, and depletion of ATP, thus demonstrating that the TRP and TRPL channels are specific targets of metabolic stress. These results shed new light on the properties of the TRP and TRPL channels by showing that a constitutive ATP-dependent process is required to keep these channels closed in the dark, a requirement that would make them sensitive to metabolic stress.
Key words: TRP and TRPL channels; Drosophila mutants; anoxia; mitochondrial uncouplers; ion-selective microelectrodes; metabolic stress
Copyright © 2000 Society for Neuroscience 0270-6474/00/20155748-08$05.00/0
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