Mitochondrial fusion regulates proliferation and differentiation in the type II neuroblast lineage in Drosophila

Optimal mitochondrial function determined by mitochondrial dynamics, morphology and activity is coupled to stem cell differentiation and organism development. However, the mechanisms of interaction of signaling pathways with mitochondrial morphology and activity are not completely understood. We assessed the role of mitochondrial fusion and fission in the differentiation of neural stem cells called neuroblasts (NB) in the Drosophila brain. Depleting mitochondrial inner membrane fusion protein Opa1 and mitochondrial outer membrane protein Marf in the Drosophila type II NB lineage led to mitochondrial fragmentation and loss of activity. Opa1 and Marf depletion did not affect the numbers of type II NBs but led to a decrease in differentiated progeny. Opa1 depletion decreased the mature intermediate precursor cells (INPs), ganglion mother cells (GMCs) and neurons by the decreased proliferation of the type II NBs and mature INPs. Marf depletion led to a decrease in neurons by a depletion of proliferation of GMCs. On the contrary, loss of mitochondrial fission protein Drp1 led to mitochondrial clustering but did not show defects in differentiation. Depletion of Drp1 along with Opa1 or Marf also led to mitochondrial clustering and suppressed the loss of mitochondrial activity and defects in proliferation and differentiation in the type II NB lineage. Opa1 depletion led to decreased Notch signaling in the type II NB lineage. Further, Notch signaling depletion via the canonical pathway showed mitochondrial fragmentation and loss of differentiation similar to Opa1 depletion. An increase in Notch signaling showed mitochondrial clustering similar to Drp1 mutants. Further, Drp1 mutant overexpression combined with Notch depletion showed mitochondrial fusion and drove differentiation in the lineage, suggesting that fused mitochondria can influence differentiation in the type II NB lineage. Our results implicate crosstalk between proliferation, Notch signaling, mitochondrial activity and fusion as an essential step in differentiation in the type II NB lineage. Author summary Mitochondrial morphology and function are coupled to stem cell differentiation and organism development. It is of interest to assess the mechanisms of interaction of mitochondrial dynamics with signaling pathways during stem cell differentiation. We have assessed the role of mitochondrial fusion and fission in the differentiation of neural stem cells called neuroblasts (NB) in the Drosophila brain. Depleting mitochondrial fusion proteins Opa1 and Marf led to mitochondrial fragmentation, loss of mitochondrial activity and proliferation, thereby causing a decrease in the numbers of differentiated cells in each type II NB lineage. Mutants in mitochondrial fission protein Drp1 led to mitochondrial fusion but did not cause any differentiation defects. Decreased Notch signaling by the canonical pathway led to mitochondrial fragmentation and a decrease in differentiated cells in each type II NB lineage. Expression of Drp1 mutants in type II NB lineages depleted of Opa1 and Marf suppressed their proliferation and differentiation defects. Expression of Drp1 mutant in type II NB lineages depleted of Notch also led to a rescue of differentiated progeny in each lineage. Our results implicate crosstalk between Notch signaling, mitochondrial activity and fusion as important steps for proliferation and differentiation in the type II NB lineage.

Automated summary

Mitochondrial morphology and function play crucial roles in stem cell differentiation and organism development. Depleting mitochondrial fusion proteins in Drosophila neuroblasts led to fragmentation and loss of activity, affecting differentiation. Mutants in mitochondrial fission protein did not affect differentiation. Disruption of Notch signaling pathway led to fragmentation, while its upregulation resulted in clustering of mitochondria. Crosstalk between proliferation, Notch signaling, mitochondrial activity, and fusion are essential for differentiation in neuroblast lineages.