Fig. 6: Mirana regulates the morphology of mitochondria in flies through expression tuning of downstream genes. | Nature Communications

Fig. 6: Mirana regulates the morphology of mitochondria in flies through expression tuning of downstream genes.

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

Fig. 6

A–E Mitochondria (labeled with mito-GFP) in DCNs axons under constitutive downregulation of Mirana target genes. F–H Temporal pink1 downregulation during development (pink1-RNAi109614/UAS-mito::GFP;ato-Gal414A,UAS-CD4::tdTomato/tub-Gal80ts vs UAS-KK30B40D/UAS-mito::GFP;ato-Gal414A,UAS-CD4::tdTomato/tub-Gal80ts) during P48-adult time window (time point following Mirana peak expression) recapitulates the elongated mitochondria phenotype. For developmental depletion of pink1, flies were kept at 18 °C until P48, then transferred to 29 °C until eclosion, and adults after hatching were transferred to 18 °C, and kept there for 10–14 days before dissection. Mitochondria size was manually measured through the Z-stack of a confocal image of one/two axons in the Optic chiasm and Medulla region in at least 3/4 different brains of each sex. Scale bar – 10 µm. I–L TransTango labeling of postsynaptic connectivity in Drp1 (Drp1-RNAi44155, n = 17 optic lobes), Atg1 (Atg1-RNAi16133, n = 20 optic lobes), and pink1 (pink1-RNAi109614, n = 18 optic lobes) downregulation in DCNs. L Number of postsynaptic cells was calculated by manually counting cell bodies in the medulla (white arrowheads) labeled with dsRed fluorescence (magenta and grayscale), including all cells with weak or strong labeling to reveal all potential connections. In case of pink1 downregulation, just like in Mirana silencing condition, we can find M-DCNs axons in Medulla (yellow arrowheads), however, TransTango labelling is severely decreased (K, K’, n = 18 optic lobes). Scale bar – 20 µm. Statistical analysis was done using the nonparametric Kruskal–Wallis test with Dunn correction for multiple comparisons and one-way ANOVA with Bonferroni correction for multiple comparisons respectively. Adjusted p values. *p < 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001, ns not significant. Schematics created in BioRender. Hassan, B. (2025) https://BioRender.com/fma66dn.

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