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. Author manuscript; available in PMC: 2018 Feb 8.
Published in final edited form as: Neuron. 2017 Jan 19;93(3):560–573.e6. doi: 10.1016/j.neuron.2016.12.017

Figure 2. Pharmacological and Genetic Inhibition of ETC and oxPhos and Its Impact on NSPC Function In Vitro and Mitochondria In Vivo.

Figure 2

(A–D) Blocking mitochondrial complex I and V by rotenone and oligomycin, respectively, leads to a significant decrease in membrane potential (A) and ATP content (B); rotenone and oligomycin treatment almost completely abolish cell proliferation (C) and increase cell death (D)

(E) Genotyping PCR of the Tfam locus of Tfamfl/fl NSPCs transduced with either a GFP-encoding control MMLV (ctrl) or an MMLV encoding for GFP and Crerecombinase (Cre) reveals recombination of the conditional Tfam locus in the context of the GFP/Cre encoding MMLV. Note the presence of the TfamloxP PCR product in GFP/Cre transduced NSPCs, which indicates that recombination of the conditional Tfam locus was present only in a subpopulation of NSPCs.

(F–I) Tfamcko NSPC cultures displayed a significant decrease in membrane potential (F) and ATP production (G) compared to control cells (ctrl); proliferation was significantly decreased (H) and cell death increased (I).

(J–M) Immunostaining for the mitochondrial proteins HSP60 (red) and Cox1 (white); GFP (green) served to identify recombined cells (outlined by white dotted line).

(L and M) Reconstruction of recombinant ctrl and Tfamcko cells. All cells contained HSP60+ mitochondria, but only in ctrl cells, mitochondria invariably co-expressed Cox1 (J and L); in Tfamcko mice, recombinant cells showed strongly diminished Cox1 expression and displayed aberrant mitochondrial morphology (K and M). Data represented as mean ± SEM; t test was performed to determine significance; all scale bars = 10 µm.