Sehgal et al. (2025). iScience. doi: 10.1016/j.isci.2025.114388
…the critical analysis of the article. Declaration of interests The authors declare no competing interests. STAR★Methods Key resources table REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-TH Pel-Freez Biologicals P40101, RRID:AB_461064 anti-elav DSHB 7E8A10, RRID:AB_528218 anti-rat-A488 Thermo Fisher Sci A-21208, RRID:AB_2535794 anti-rabbit-A647 Thermo Fisher Sci A-27040, RRID:AB_2536101 Chemicals, peptides,…
Pel-Freez products cited: Donkey Serum, Sterile, Non-Hemolyzed (39125), Rabbit Anti-Tyrosine Hydroxylase (P40101)
Abstract
The dopaminergic system has a large role in behavior and neurological disease, and understanding dopamine level regulation <i>in vivo</i> is critical. To identify dopamine regulators, we utilized <i>Drosophila melanogaster</i> cuticle pigmentation, where dopamine is a precursor to melanin. We measured dopamine from known pigmentation mutants (e.g., <i>tan</i>, <i>ebony</i>, <i>black</i>) 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 <i>Tyrosine hydroxylase (TH)</i>, suggesting dopamine levels are tightly regulated in the nervous system. We pursued two hits that reduced brain dopamine levels, <i>clueless</i> and <i>mask</i>. Further examination suggests that the <i>mask</i> regulates the transcription of <i>TH</i> 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.