RNAi-based screen for pigmentation in Drosophila melanogaster reveals regulators of brain dopamine and sleep

利用RNAi技术筛选果蝇色素沉着相关基因,揭示了大脑多巴胺和睡眠的调节因子

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Abstract

The dopaminergic system has a large role in behavior and neurological disease, and understanding dopamine level regulation in vivo is critical. To identify dopamine regulators, we utilized Drosophila melanogaster cuticle pigmentation, where dopamine is a precursor to melanin. We measured dopamine from known pigmentation mutants (e.g., tan, ebony, black) and performed an RNAi-based screen to identify additional regulators. We found 153 hits, enriched for developmental signaling pathways and mitochondria-associated proteins. From 35 prioritized candidates, 11 affected head dopamine levels. Effects on brain dopamine were mild, even knocking down the rate-limiting synthesis enzyme Tyrosine hydroxylase (TH), suggesting dopamine levels are tightly regulated in the nervous system. We pursued two hits that reduced brain dopamine levels, clueless and mask. Further examination suggests that the mask regulates the transcription of TH and affects dopamine-dependent sleep. In summary, studying genes that affect cuticle pigmentation helped to identify genes that alter dopamine metabolism and a behavioral regulator.

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