Orai mediated Calcium entry sets the excitability threshold of central dopaminergic neurons by regulation of gene expression

![Figure][1] Graphical Abstract Maturation and fine-tuning of neural circuits frequently requires neuromodulatory signals that set the excitability threshold, neuronal connectivity and synaptic strength. Here we present a mechanistic study of how neuromodulator stimulated intracellular Ca2+ signals, through the store – operated Ca2+ channel Orai, regulate intrinsic neuronal properties by control of developmental gene expression in flight promoting central dopaminergic neurons (fpDANs). The fpDANs receive cholinergic inputs for release of dopamine at a central brain tripartite synapse that sustains flight ([Sharma and Hasan, 2020][2]). Cholinergic inputs act on the muscarinic acetylcholine receptor to stimulate intracellular Ca2+ release through the endoplasmic reticulum (ER) localised inositol 1,4,5-trisphosphate receptor followed by ER-store depletion and Orai mediated store-operated Ca2+ entry (SOCE). Analysis of gene expression in fpDANs followed by genetic, cellular and molecular studies identified Orai-mediated Ca2+ entry as a key regulator of excitability in fpDANs during circuit maturation. SOCE activates the transcription factor Trithorax-like (Trl) which in turn drives expression of a set of genes including Set2 , that encodes a histone 3 Lysine 36 methyltransferase (H3K36me3). Set2 function establishes a positive feedback loop, essential for receiving neuromodulatory cholinergic inputs and sustaining SOCE. Chromatin modifying activity of Set2 changes the epigenetic status of fpDANs and drives expression of key ion channel and signaling genes thus setting the excitability threshold that determines dopamine release for maintenance of long flight. 1. Store-operated Ca2+ entry (SOCE) through Orai is required in a set of flight-promoting central dopaminergic neurons (fpDANs) during late pupae and early adults to establish their gene expression profile. 2. SOCE activates a homeobox transcription factor, ‘ Trithorax-like ’ and thus regulates expression of histone modifiers Set2 and E(z) to generate a balance between opposing epigenetic signatures of H3K36me3 and H3K27me3 on downstream genes. 3. SOCE drives a transcriptional feedback loop to ensure expression of key genes required for neuronal function including the muscarinic acetylcholine receptor ( mAChR ) and the inositol 1,4,5-trisphosphate receptor ( itpr) . 4. The transcriptional program downstream of SOCE is key to functional maturation of the dopaminergic neurons, enabling their neuronal excitability and synaptic transmission required for adult flight. ### Competing Interest Statement The authors have declared no competing interest. [1]: pending:yes [2]: #ref-87