Drosophila glutathione S-transferase S1 exacerbates phenotypes of the gain-of-function voltage-gated sodium channel mutant para Shu through a hemocyte-mediated mechanism

Drosophila glutathione S-transferase S1 (GstS1) is a unique member of the glutathione S-transferase family and is considered the ortholog of vertebrate hematopoietic prostaglandin D synthase based on amino acid sequence similarity. A previous forward genetic screen revealed that a null mutation in GstS1 dominantly suppresses genetically induced behavioral hyperexcitability in several seizure-prone Drosophila mutants, including para Shu , a gain-of-function mutant of the voltage-gated sodium channel gene paralytic ( para ), and eas 2 , a loss-of-function mutant of the ethanolamine kinase gene easily shocked ( eas ). Here, using the GAL4/UAS system combined with GstS1 RNAi, we show that hemocyte-specific GstS1 knockdown suppresses hyperexcitable phenotypes in both para Shu and eas 2 . Notably, suppression of adult para Shu phenotypes required GstS1 knockdown before adult eclosion, suggesting that GstS1 modulates adult neuronal excitability by influencing neural development through neuroimmune interactions. Consistent with this model, hemocyte-specific GstS1 knockdown rescued abnormal dendritic morphology in developing class IV dendritic arborization (C4da) neurons of para Shu larvae. Moreover, para Shu and eas 2 larvae exhibited increased numbers of sessile hemocytes surrounding C4da neuron somata, a phenotype reversed by either GstS1 knockdown or dietary supplementation with the ω-3 polyunsaturated fatty acid α-linolenic acid. Together, these findings reveal a previously unrecognized developmental role for GstS1 in promoting nervous system hyperexcitability through hemocyte-mediated neuroimmune interactions, potentially involving bioactive lipid signaling. Our results highlight neuroimmune crosstalk as an important mechanism linking genetic perturbations to altered neural development and seizure-associated hyperexcitability.