Amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) are fatal neurodegenerative disorders with significant clinical and molecular overlaps. Critical events in their pathogenesis involve cytoplasmic mislocalization and loss of nuclear functions of TAR DNA-binding protein 43 (TDP-43). Notably, unconventional metabolic conditions like type 2 diabetes mellitus and dyslipidemia paradoxically correlate with a better prognosis in ALS and FTLD, hinting at potential metabolic pathways in TDP-43-associated neurodegeneration. To better understand the interplay between TDP-43 and metabolic signalling in neurodegeneration contexts, we simulated TDP-43 loss of function via RNA interference in mouse NSC34 motor neurons, N2A neuroblastoma cells, and BV2 microglia. This was followed by a comprehensive metabolic profiling of these cellular models including metabolic flux analyses for examination of glycolysis and oxidative phosphorylation dynamics. Our findings revealed distinct cell-specific metabolic phenotypes following TDP-43 depletion. NSC34 motor neurons exhibited a hypermetabolic phenotype with accentuation of both glycolysis and oxidative phosphorylation. However, N2A cells displayed a hypometabolic phenotype, whereas BV2 microglia cells only exhibited an increase in glycolysis. These metabolic maladaptations upon TDP-43 depletion underscore the role of TDP-43 in neuronal and glial energy metabolism and provide insight into the selective vulnerability of motor neurons in TDP-43 proteinopathies. Furthermore, we have identified the dysregulated activity of the cellular energy sensor, AMP-activated protein kinase (AMPK) as a potential driver of increased glucose metabolism in motor neurons after TDP-43 depletion. Ongoing investigations focus on in vivo validation in a TDP-43 nuclear loss of function mouse model and human postmortem tissue.