Endoplasmic reticulum Ca(2+) release through ryanodine and IP(3) receptors contributes to neuronal excitotoxicity.
Overactivation of ionotropic glutamate receptors induces a Ca(2+) overload into the cytoplasm that leads neurons to excitotoxic death, a process that has been linked to several neurodegenerative disorders. While the role of mitochondria and its involvement in excitotoxicity have been widely studied, the contribution of endoplasmic reticulum (ER), another crucial intracellular store in maintaining Ca(2+) homeostasis, is not fully understood. In this study, we analyzed the contribution of ER-Ca(2+) release through ryanodine (RyR) and IP(3) (IP(3)R) receptors to a neuronal in vitro model of excitotoxicity. NMDA induced a dose-dependent neuronal death, which was significantly decreased by ER-Ca(2+) release inhibitors in cortical neurons as well as in organotypic slices. Furthermore, ryanodine and 2APB, RyR and IP(3)R inhibitors respectively, attenuated NMDA-triggered intracellular Ca(2+) increase and oxidative stress, whereas 2APB reduced mitochondrial membrane depolarization and caspase-3 cleavage. Consistent with ER-Ca(2+) homeostasis disruption, we observed that NMDA-induced ER stress, characterized here by eIF2alpha phosphorylation and over-expression of GRP chaperones which were regulated by ER-Ca(2+) release inhibitors. These results demonstrate that Ca(2+) release from ER contributes to neuronal death by both promoting mitochondrial dysfunction and inducing specific stress and apoptosis pathways during excitotoxicity.
Ruiz
Asier
A
Centro de Investigación Biomédica en Red en Enfermedades Neurodegenerativas (CIBERNED), Spain.
Matute
Carlos
C
Alberdi
Elena
E
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001896:Boron Compounds / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism* / Q000633:toxicity*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000378:metabolism / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D004721:Endoplasmic Reticulum / Q000187:drug effects* / Q000378:metabolism*; D000091342:Endoplasmic Reticulum Chaperone BiP; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000235:genetics; D066298:In Vitro Techniques; D053496:Inositol 1,4,5-Trisphosphate Receptors / Q000494:pharmacology*; D008928:Mitochondria / Q000187:drug effects / Q000473:pathology; D016202:N-Methylaspartate / Q000633:toxicity; D009474:Neurons / Q000378:metabolism / Q000473:pathology*; D018384:Oxidative Stress / Q000187:drug effects; D051381:Rats; D012433:Ryanodine / Q000494:pharmacology*
D001896:Boron Compounds; D000091342:Endoplasmic Reticulum Chaperone BiP; D015852:Eukaryotic Initiation Factor-2; D006360:Heat-Shock Proteins; D053496:Inositol 1,4,5-Trisphosphate Receptors; D012433:Ryanodine; D016202:N-Methylaspartate; C109986:2-aminoethoxydiphenyl borate; D002118:Calcium