A new procedure for amyloid β oligomers preparation enables the unambiguous testing of their effects on cytosolic and mitochondrial Ca2+ entry and cell death in primary neurons

NEUROSCIENCE LETTERS

卷:612

Article

Caballero, Erica¹  Calvo-Rodriguez, Maria¹  Gonzalo-Ruiz, Alicia²  Villalobos, Carlos¹  Nunez, Lucia^1,3 

[1] CSIC, Inst Mol Biol & Genet IBGM, Valladolid, Spain

[2] Univ Valladolid, Inst Neurosci Castilla & Leon, Lab Neuroanat, Soria Campus, E-47005 Valladolid, Spain

[3] Univ Valladolid, Sch Med, Dept Biochem & Mol Biol & Physiol, C Ramon Cajal, E-47005 Valladolid, Spain

关键词: Calcium;    Mitochondria;    Alzheimer's disease;    Amyloid beta peptide;    Oligomers;   

DOI  :  10.1016/j.neulet.2015.11.041

来源: Elsevier

【 摘 要 】

Oligomers of the amyloid beta peptide (A beta(o)) are becoming the most likely neurotoxin in Alzheimer's disease. Controversy remains on the mechanisms involved in neurotoxicity induced by A beta(o) and the targets involved. We have reported that A beta(o) promote Ca2+ entry, mitochondrial Ca2+ overload and apoptosis in cultured cerebellar neurons. However, recent evidence suggests that some of these effects could be induced by glutamate receptor agonists solved in F12, the media in which A beta(o) are prepared. Here we have tested the effects of different media on A beta(o) formation and on cytosolic Ca2+ concentration ([Ca2+](cyt)) in rat cerebellar and hippocampal cell cultures. We found that A beta(o) prepared according to previous protocols but solved in alternative media including saline, MEM and DMEM do not allow oligomer formation and fail to increase [Ca2+](cyt). Changes in the oligomerization protocol and supplementation of media with selected salts reported to favor oligomer formation enable A beta(o) formation. A beta(o) prepared by the new procedure and containing small molecular weight oligomers increased [Ca2+](cyt), promoted mitochondrial Ca2+ overload and cell death in cerebellar granule cells and hippocampal neurons. These results foster a role for Ca2+ entry in neurotoxicity induced by A beta(o) and provide a reliable procedure for investigating the Ca2+ entry pathway promoted by A beta(o). (C) 2015 Elsevier Ireland Ltd. All rights reserved.

【 授权许可】

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