Fig. 7: Overexpression of pink1 in DCNs rescues all Mirana loss of function phenotypes. | Nature Communications

Fig. 7: Overexpression of pink1 in DCNs rescues all Mirana loss of function phenotypes.

From: Temporal transcriptional regulation of mitochondrial morphology primes activity-dependent circuit connectivity

Fig. 7

A–C’ Mitochondria size (labeled with mito-GFP) in DCNs axons was rescued to that of controls (w1118;UAS-mito::GFP/UAS-KK40D30B,ato-Gal414A,UAS-CD4::tdTomato/UAS-pink1, A–A’) with pink1 overexpression (w1118;UAS-mito::GFP/ UAS-Mirana-RNAi100127,ato-Gal414A,UAS-CD4::tdTomato/UAS-pink1, C–C’). Mitochondria size was manually measured through the Z-stack of a confocal image of one/two axons in the optic chiasm and medulla in at least 3/4 different brains of each sex (D). Scale bar – 10 µm. H–G’ Anterograde transsynaptic labeling of postsynaptic partner cells using the TransTango approach (E’, G’, n = 22 optic lobes for w1118;UAS-mito::GFP/UAS-KK40D30B,ato-Gal414A,UAS-CD4::tdTomato/UAS-pink1 and 20 optic lobes for w1118;UAS-mito::GFP/ UAS-Mirana-RNAi100127,ato-Gal414A,UAS-CD4::tdTomato/UAS-pink1). Postsynaptic connectivity was calculated by manually counting cell bodies in Medulla (white arrowheads) labeled with dsRed fluorescence (magenta and grayscale) (H, n = 17 optic lobes for Control-RNAi30B40D. N = 20 optic lobes for Mirana-RNAi100127. N = 22 optic lobes for Control-RNAi30B40D,UAS-pink1. N = 20 optic lobes for Mirana-RNAi100127,UAS-pink1), including all cells with weak or strong labeling to reveal all potential connections. Data are presented as mean values +/− SEM. Scale bar – 20 µm. I–M Number of DCN axons reaching the Medulla in adult flies. pink1 overexpression (ato-Gal414A > UAS-Mirana-RNAi100127,UAS-pink1) restored the M-DCNs axons, caused by Mirana downregulation (M, n = 33 optic lobes for Control. N = 16 optic lobes for Mirana-CRISPR/Cas9, n = 21 optic lobes for pink1-RNAi n = 26 optic lobes for Control,UAS-pink1, n = 40 optic lobes for Mirana-RNAi100127,UAS-pink1. Measurements were done equally in flies of both sexes. Data are presented as mean values ± SEM. Me medulla. Scale bar − 20 μm. Statistical analysis was done using one-way ANOVA with Dunnett, and Bonferroni corrections for multiple comparisons in mitochondria length comparison. Adjusted p values. *p < 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001, ns not significant.

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