Mitochondrial fusion regulates proliferation and differentiation in the type II neuroblast lineage in Drosophila
Fig 6
Analysis of mitochondrial morphology in Notch mutants and Notch signaling in Opa1 depleted type II NB lineages.
A:B: Representative confocal images of type II NB lineages (A) showing reduced numbers of mature INPs in notch RNAi. Quantification of mature INPs (B) in Control (16 type II NB lineages, 4 brains), NotchFL (15,5), notch RNAi (5,3). C-E: Type II NBs (mCD8-GFP, green, yellow dotted line) showing Notch signaling mediated regulation of mitochondrial morphology (red) by ATPβ antibody using STED microscopy (C). +/+ (100% tubular, 75 NBs, 22 Brains), NotchFL (68% clustered, 103,16), Nintra (76% clustered, 58,6), notch RNAi (50% fragmented, 30,14), su(H) RNAi (80% fragmented, 23,12). Graph shows the distribution of mitochondria into fragmented, intermediate and clustered in the form of a stacked histogram (D). The percentage documented on each bar in the histogram is for the group that is seen at the maximum extent. Average mitochondrial area quantification (E) in +/+ (9 Type II NBs,3 brains), NotchFL (22,5), Nintra (15,4), notch RNAi (16,4), su(h) RNAi (17,4). Scale bar- 5μm. F-H: Type II NBs (mCD8-GFP, green, yellow dotted line) stained for NICD (F) and observed for distribution in +/+, Drp1SD, opa1 RNAi, Drp1SD;opa1 RNAi, marf RNAi, Drp1SD;marf RNAi. Quantification of NICD in type II NBs (G) in +/+ (19,10), opa1 RNAi (20,12), marf RNAi (15,12), Drp1SD (22,5), Drp1SD;opa1 RNAi (28,6), Drp1SD;marf RNAi (11,5). Quantification of membrane NICD intensities in lineage (H, yellow arrows) in +/+ (6 lineages,3 brains), Drp1SD (10,4), opa1 RNAi (9,3), marf RNAi (9,3), Drp1SD;opa1 RNAi (9,4), Drp1SD;marf RNAi (8,4). Scale bar- 10μm. I: Representative confocal images of type II NB lineages showing expression of E(spl)mγ-GFP (green) in type II NB (large Dpn+ dotted large circles) and mature INPs (small Dpn+ dotted small circles). B, D, F & G: Graphs show mean ± sd. Statistical analysis was done by using unpaired t-test. ns- non significant, *- p<0.1, **- p<0.01, ***- p<0.001.