Genrikhs Elisaveta E, Stelmashook Elena V, Golyshev Sergey A, Aleksandrova Olga P, Isaev Nickolay K
Research Center of Neurology, Volokolamskoe Shosse 80, 125367 Moscow, Russia.
Research Center of Neurology, Volokolamskoe Shosse 80, 125367 Moscow, Russia.
Brain Res Bull. 2017 Apr;130:90-94. doi: 10.1016/j.brainresbull.2017.01.004. Epub 2017 Jan 6.
Streptozotocin (STZ) is a glucosamine-nitrosourea compound used for experimental simulation of sporadic Alzheimer's disease at intracerebroventricular administration in vivo. The studies of STZ influence on neurons of central nervous system performed on the primary cultures are practically absent. We have shown the application of STZ (1-5mM) in primary culture for 48h induced strong dose-dependent death in cultured cerebellar granule neurons. This toxic effect was decreased by pyruvate, insulin partially. Using the indicator Fluo-4 AM for measurements of intracellular calcium ions and tetramethylrhodamine ethyl ester (TMRE) for detection of changes of mitochondrial membrane potential in live cells we have shown that 5 h-exposure to STZ induced intensive increase of Fluo-4 and decrease TMRE fluorescence in neurons. STZ exposure caused considerable ultrastructural alterations in granule neurons: chromatin clumping, swelling of the endoplasmic reticulum and mitochondria, and disruption of the mitochondrial cristae. Probably, STZ significantly impaired glucose metabolism and mitochondrial function that, in turn, resulted in mitochondrial membrane potential damage, excessive calcium overload and neuronal death.
链脲佐菌素（STZ）是一种氨基葡萄糖亚硝基脲化合物，用于在体内脑室内给药时对散发性阿尔茨海默病进行实验模拟。关于STZ对原代培养的中枢神经系统神经元影响的研究实际上并不存在。我们已经表明，在原代培养中应用1-5mM的STZ 48小时会在培养的小脑颗粒神经元中诱导强烈的剂量依赖性死亡。丙酮酸和胰岛素可部分降低这种毒性作用。使用Fluo-4 AM指示剂测量活细胞内的钙离子，并用四甲基罗丹明乙酯（TMRE）检测线粒体膜电位的变化，我们发现，暴露于STZ 5小时会导致神经元中Fluo-4强烈增加，TMRE荧光降低。STZ暴露导致颗粒神经元出现明显的超微结构改变：染色质凝聚、内质网和线粒体肿胀以及线粒体嵴破坏。可能是STZ显著损害了葡萄糖代谢和线粒体功能，进而导致线粒体膜电位损伤、钙过载过多和神经元死亡。
