L-Lactate-Mediated Neuroprotection against Glutamate-Induced Excitotoxicity Requires ARALAR/AGC1.
ARALAR/AGC1/Slc25a12, the aspartate-glutamate carrier from brain mitochondria, is the regulatory step in the malate-aspartate NADH shuttle, MAS. MAS is used to oxidize cytosolic NADH in mitochondria, a process required to maintain oxidative glucose utilization. The role of ARALAR was analyzed in two paradigms of glutamate-induced excitotoxicity in cortical neurons: glucose deprivation and acute glutamate stimulation. ARALAR deficiency did not aggravate glutamate-induced neuronal death in vitro, although glutamate-stimulated respiration was impaired. In contrast, the presence of L-lactate as an additional source protected against glutamate-induced neuronal death in control, but not ARALAR-deficient neurons.l-Lactate supplementation increased glutamate-stimulated respiration partially prevented the decrease in the cytosolic ATP/ADP ratio induced by glutamate and substantially diminished mitochondrial accumulation of 8-oxoguanosine, a marker of reactive oxygen species production, only in the presence, but not the absence, of ARALAR. In addition,l-lactate potentiated glutamate-induced increase in cytosolic Ca(2+), in a way independent of the presence of ARALAR. Interestingly,in vivo, the loss of half-a-dose of ARALAR in aralar(+/-)mice enhanced kainic acid-induced seizures and neuronal damage with respect to control animals, in a model of excitotoxicity in which increased L-lactate levels and L-lactate consumption have been previously proven. These results suggest that,in vivo, an inefficient operation of the shuttle in the aralar hemizygous mice prevents the protective role of L-lactate on glutamate excitotoxiciy and that the entry and oxidation of L-lactate through ARALAR-MAS pathway is required for its neuroprotective function.


CONCLUSIONS
Lactate now stands as a metabolite necessary for multiple functions in the brain and is an alternative energy source during excitotoxic brain injury. Here we find that the absence of a functional malate-aspartate NADH shuttle caused by aralar/AGC1 disruption causes a block in lactate utilization by neurons, which prevents the protective role of lactate on excitotoxicity, but not glutamate excitotoxicity itself. Thus, failure to use lactate is detrimental and is possibly responsible for the exacerbated in vivo excitotoxicity in aralar(+/-)mice.
Departamento de Biología Molecular, Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid, Madrid 28049, Spain, Centro de Investigación Biomédica en Red de Enfermedades Raras, Madrid 28029, Spain, and Instituto de Investigación Sanitaria Fundación Jiménez Díaz, Madrid 28006, Spain.
Irene
I
Llorente-Folch
Departamento de Biología Molecular, Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid, Madrid 28049, Spain, Centro de Investigación Biomédica en Red de Enfermedades Raras, Madrid 28029, Spain, and Instituto de Investigación Sanitaria Fundación Jiménez Díaz, Madrid 28006, Spain.
Carlos B
CB
Rueda
Departamento de Biología Molecular, Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid, Madrid 28049, Spain, Centro de Investigación Biomédica en Red de Enfermedades Raras, Madrid 28029, Spain, and Instituto de Investigación Sanitaria Fundación Jiménez Díaz, Madrid 28006, Spain.
Irene
I
Pérez-Liébana
Departamento de Biología Molecular, Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid, Madrid 28049, Spain, Centro de Investigación Biomédica en Red de Enfermedades Raras, Madrid 28029, Spain, and Instituto de Investigación Sanitaria Fundación Jiménez Díaz, Madrid 28006, Spain.
Jorgina
J
Satrústegui
Departamento de Biología Molecular, Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid, Madrid 28049, Spain, Centro de Investigación Biomédica en Red de Enfermedades Raras, Madrid 28029, Spain, and Instituto de Investigación Sanitaria Fundación Jiménez Díaz, Madrid 28006, Spain bpardo@cbm.csic.es.
Beatriz
B
Pardo
D053674:Aggrecans / Q000172:deficiency*; D000818:Animals; D002478:Cells, Cultured; D018698:Glutamic Acid / Q000633:toxicity*; D019344:Lactic Acid / Q000378:metabolism* / Q000494:pharmacology; D008297:Male; D051379:Mice; D057507:Mice, 129 Strain; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D033681:Mitochondrial Membrane Transport Proteins / Q000172:deficiency*; D000066829:Neuroprotection / Q000187:drug effects* / Q000502:physiology
C508320:Acan protein, mouse; D053674:Aggrecans; D033681:Mitochondrial Membrane Transport Proteins; C491956:Slc25a12 protein, mouse; D019344:Lactic Acid; D018698:Glutamic Acid
ARALAR/AGC1; glutamate excitotoxicity; kainic acid; l-lactate; malate-aspartate NADH shuttle; neuroprotection