Inhibition of Mitochondrial Complex III Blocks Neuronal Differentiation and Maintains Embryonic Stem Cell Pluripotency

Abstract

The mitochondrion is emerging as a key organelle in stem cell biology, acting as a regulator of stem cell pluripotency and differentiation. In this study we sought to understand the effect of mitochondrial complex III inhibition during neuronal differentiation of mouse embryonic stem cells. When exposed to antimycin A, a specific complex III inhibitor, embryonic stem cells failed to differentiate into dopaminergic neurons, maintaining high Oct4 levels even when subjected to a specific differentiation protocol. Mitochondrial inhibition affected distinct populations of cells present in culture, inducing cell loss in differentiated cells, but not inducing apoptosis in mouse embryonic stem cells. A reduction in overall proliferation rate was observed, corresponding to a slight arrest in S phase. Moreover, antimycin A treatment induced a consistent increase in HIF-1α protein levels. The present work demonstrates that mitochondrial metabolism is critical for neuronal differentiation and emphasizes that modulation of mitochondrial functions through pharmacological approaches can be useful in the context of controlling stem cell maintenance/differentiation.

Mitochondria Regulate the Differentiation of Stem Cells from Human Exfoliated Deciduous Teeth

“…In contrast, lactate levels are increased during this period (Prigione et al, 2010). In addition, inhibition of mitochondrial activity by rotenone enhances the reprogramming of somatic cells into iPSCs (Son et al, 2013), and the inhibition of mitochondrial activity by antimycine A increases Oct-4 expression (Pereira et al, 2013). Taken together, these data indicate that low levels of mitochondrial activity tend to maintain stem cells in the undifferentiated state.…”

“…Our results were similar to the results of previous studies of the neuronal differentiation of ESCs. Similar to SHED, ESCs also exhibit increased mitochondrial activity during differentiation (Pereira et al, 2013). Furthermore, the inhibition of mitochondrial activity also reduces the neuronal differentiation of ESCs (Pereira et al, 2013).…”

“…Rotenone increases the production of mtROS (Cadenas et al, 1977), and mtROS production reduces the neuronal differentiation of ESCs (Pereira et al, 2013). mtROS production increased when SHED were cultured with rotenone (Fig.…”

“…Similar to SHED, ESCs also exhibit increased mitochondrial activity during differentiation (Pereira et al, 2013). Furthermore, the inhibition of mitochondrial activity also reduces the neuronal differentiation of ESCs (Pereira et al, 2013). Interestingly, to maintain their stemness, ESCs harbor a limited number of mitochondria and reduce their mitochondrial activity before differentiation (Armstrong et al, 2010;Chen et al, 2012).…”

“…Inhibition of mitochondrial complex III, a complex required for oxidative phosphorylation, blocks the neuronal differentiation of ESCs (Pereira et al, 2013). Furthermore, inhibition of mitochondrial complex III results in high expression of Oct-4 during the neuronal differentiation of ESCs (Pereira et al, 2013).…”