Villalobos Carlos, Caballero Erica, Sanz-Blasco Sara, Núñez Lucía
Instituto de Biología y Genética Molecular (IBGM), Universidad de Valladolid and Consejo Superior de Investigaciones Científicas (CSIC), Valladolid, Spain.
Methods Mol Biol. 2012;849:289-302. doi: 10.1007/978-1-61779-551-0_20.
Neurotoxicity in Alzheimer's disease (AD) is associated to dishomeostasis of intracellular Ca(2+) induced by amyloid β peptide (Aβ) species. Understanding of the effects of Aβ on intracellular Ca(2+) homeostasis requires preparation of the different Aβ assemblies including oligomers and fibrils and the testing of their effects on cytosolic and mitochondrial Ca(2+) in neurons. Procedures for cerebellar granule cell culture, preparation of Aβ species as well as fluorescence and bioluminescence imaging of cytosolic and mitochondrial Ca(2+) in neurons are described.
阿尔茨海默病(AD)中的神经毒性与淀粉样β肽(Aβ)诱导的细胞内Ca(2+)失衡有关。要了解Aβ对细胞内Ca(2+)稳态的影响,需要制备包括寡聚体和原纤维在内的不同Aβ聚集体,并测试它们对神经元胞质和线粒体Ca(2+)的影响。本文描述了小脑颗粒细胞培养、Aβ聚集体制备以及神经元胞质和线粒体Ca(2+)的荧光和生物发光成像的方法。