MLKL overexpression leads to Ca2+ and metabolic dyshomeostasis in a neuronal cell model

Cell Calcium. 2024 May:119:102854. doi: 10.1016/j.ceca.2024.102854. Epub 2024 Feb 6.

Abstract

The necroptotic effector molecule MLKL accumulates in neurons over the lifespan of mice, and its downregulation has the potential to improve cognition through neuroinflammation, and changes in the abundance of synaptic proteins and enzymes in the central nervous system. Notwithstanding, direct evidence of cell-autonomous effects of MLKL expression on neuronal physiology and metabolism are lacking. Here, we tested whether the overexpression of MLKL in the absence of cell death in the neuronal cell line Neuro-2a recapitulates some of the hallmarks of aging at the cellular level. Using genetically-encoded fluorescent biosensors, we monitored the cytosolic and mitochondrial Ca2+ levels, along with the cytosolic concentrations of several metabolites involved in energy metabolism (lactate, glucose, ATP) and oxidative stress (oxidized/reduced glutathione). We found that MLKL overexpression marginally decreased cell viability, however, it led to reduced cytosolic and mitochondrial Ca2+ elevations in response to Ca2+ influx from the extracellular space. On the contrary, Ca2+ signals were elevated after mobilizing Ca2+ from the endoplasmic reticulum. Transient elevations in cytosolic Ca2+, mimicking neuronal stimulation, lead to higher lactate levels and lower glucose concentrations in Neuro-2a cells when overexpressing MLKL, which suggest enhanced neuronal glycolysis. Despite these alterations, energy levels and glutathione redox state in the cell bodies remained largely preserved after inducing MLKL overexpression for 24-48 h. Taken together, our proof-of-concept experiments are consistent with the hypothesis that MLKL overexpression in the absence of cell death contributes to both Ca2+ and metabolic dyshomeostasis, which are cellular hallmarks of brain aging.

Keywords: Aging; Calcium; Lactate; MLKL; Metabolism; Mitochondria; Necroptosis; Neuron.

MeSH terms

  • Animals
  • Cell Death
  • Cell Line
  • Glucose / metabolism
  • Lactates* / metabolism
  • Lactates* / pharmacology
  • Mice
  • Neurons* / metabolism
  • Protein Kinases / metabolism

Substances

  • Lactates
  • Glucose
  • MLKL protein, mouse
  • Protein Kinases