Journal: iScience
Article Title: Dopaminergic neurons are vulnerable to dysregulation of YEATS2-dependent calcium homeostasis
doi: 10.1016/j.isci.2026.115855
Figure Lengend Snippet: Pharmacological attenuation of SOCE restores DAergic synaptic integrity in dYEATS2 -deficient flies (A) Experimental scheme. Flies expressing membrane-tethered mCD8-GFP and dYEATS2 RNAi specifically in dopaminergic neurons ( Ddc>mCD8-GFP>dYEATS2-IR ) were reared on standard medium supplemented with vehicle or the Orai inhibitor BTP2 (YM-58483) at 1 μM or 10 μM. Adult flies were transferred to fresh vials containing the same treatments, and heads were dissected at 5 days post-eclosion for confocal imaging or RNA extraction. (B) Representative confocal images (posterior→anterior orientation) showing functionally active DAergic neurons identified by co-localization of mCD8-GFP (membrane marker expressed under Ddc-GAL4 driver) and tyrosine hydroxylase (TH) immunoreactivity. Central brain boundaries are indicated by dotted lines; major DA clusters (PAL, PAM, PPL1, PPM3) are highlighted with red dashed circles. Scale bars, 250μm. (C) Quantification of EGFP-TH co-localization (number of co-localized puncta) was performed on 10 independent brains per condition using the colocalization module in CellSense (Olympus). Bars show mean ± SEM; BTP2 treatment at both 1 μM and 10 μM significantly increased the number of EGFP-TH co-localizing spots relative to untreated dYEATS2 -IR animals. (D) Transcript levels of selected dYEATS2 -responsive genes ( Gαq , trpL , vMAT , and DD2R ) were measured from dissected adult heads following vehicle or BTP2 treatment to assess whether SOCE inhibition modulates these transcriptional changes. Gene expression was determined by reverse transcription quantitative PCR (RT-qPCR). Data are presented as mean ± SEM. Statistical significance was assessed by one-way ANOVA with Šidák’s and Tukey’s post hoc tests, respectively; p < 0.05 was considered significant.
Article Snippet: Scale bars, 250μm. (C) Quantification of EGFP-TH co-localization (number of co-localized puncta) was performed on 10 independent brains per condition using the colocalization module in CellSense (Olympus).
Techniques: Expressing, Membrane, Imaging, RNA Extraction, Marker, Inhibition, Gene Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative RT-PCR
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