Exploring Excitotoxicity and Regulation of a Constitutively Active TRP Ca2+ Channel in Drosophila.

Abstract

Unregulated Ca²⁺ influx affects intracellular Ca²⁺ homoeostasis, which may lead to neuronal death. In Drosophila, following the activation of rhodopsin the TRP Ca²⁺ channel is open to mediate the light-dependent depolarization. A constitutively active TRP channel triggers the degeneration of Trp^P365 /+ photoreceptors. To explore retinal degeneration, we employed a multidisciplinary approach including live imaging using GFP tagged actin and arrestin 2. Importantly, we demonstrate that the major rhodopsin (Rh1) was greatly reduced before the onset of rhabdomere degeneration; a great reduction of Rh1 affects the maintenance of rhabdomere leading to degeneration of photoreceptors. Trp^P365 /+ also led to the up-regulation of CaMKII, which is beneficial as suppression of CaMKII accelerated retinal degeneration. We explored the regulation of TRP by investigating the genetic interaction between Trp^P365 /+ and mutants affecting the turnover of diacylglycerol (DAG). We show a loss of phospholipase C in norpA^P24 exhibited a great reduction of the DAG content delayed degeneration of Trp^P365 /+ photoreceptors. In contrast, knockdown or mutations in DAG lipase (InaE) that is accompanied by slightly reduced levels of most DAG but an increased level of DAG 34:1, exacerbated retinal degeneration of Trp^P365 /+. Together, our findings support the notion that DAG plays a role in regulating TRP. Interestingly, DAG lipase is likely required during photoreceptor development as Trp^P365 /+; inaE^N125 double mutants contained severely degenerated rhabdomeres.