Exploring Excitotoxicity and Regulation of a Constitutively Active TRP Ca(2+) Channel in Drosophila

探索果蝇中组成型活性TRP Ca(2+)通道的兴奋性毒性和调控

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Abstract

Unregulated Ca(2+) influx affects intracellular Ca(2+) homoeostasis, which may lead to neuronal death. In Drosophila, following the activation of rhodopsin the TRP Ca(2+) 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.

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