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bioRxiv · 10.64898/2026.03.25.714145

Stage-resolved iPSC-to-motoneuron differentiation: Metabolic switch & mitochondrial remodeling

Jbeily, J.·Raic, A.·Hafner, M.·Rudolf, R.
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Abstract

Development of motoneurons from stem cells is characterized by a change from glycolytic to oxidative metabolism. Since this transition remains poorly understood, we examined it at five distinct differentiation stages from hiPSC to motoneuron. While a direct comparison of hiPSCs and mature motoneurons confirmed the expected glycolytic-to-oxidative shift, the intermediate stages showed that the conversion was not monotonic. After an initial drop of glycolysis at the hiPSC-to-neuroepithelial transition, late neuroepithelial cells showed intermittent peaks of the glycolytic marker lactate dehydrogenase A and the metabolic regulator TIGAR. Furthermore, the lactate-produced-to-glucose-consumed ratio remained elevated. A fully oxidative phenotype was only assumed upon progress from neural progenitors to motoneurons, portrayed by a definitive drop of the lactate-produced-to-glucose-consumed ratio, an increase of mitochondrial membrane charging, and shifts from lactate dehydrogenase A to B, from pyruvate dehydrogenase to anaplerotic pyruvate carboxylase, and from Mitofusin 1 to 2. Together, our data show that metabolic maturation in human motoneurons does not occur as a simple switch. Instead, it unfolds through distinct stages in a directional yet nonlinear manner.

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Jbeily, J., Raic, A., Hafner, M., Rudolf, R.. 2026-03-30. Stage-resolved iPSC-to-motoneuron differentiation: Metabolic switch & mitochondrial remodeling. https://doi.org/10.64898/2026.03.25.714145

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