Oxidative stress, nitric oxide, and the mechanisms of cell death in Lurcher Purkinje cells
Oxidative stress, nitric oxide, and the mechanisms of cell death in Lurcher Purkinje cells
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DOI：
10.1002/dneu.20391
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发表时间：
2007-07-01
影响因子：
3
通讯作者：
Vogel, Michael W.
中科院分区：
文献类型：
--
作者：
McFarland, Rebecca;Blokhin, Andrei;Vogel, Michael W.
氧化应激被认为在许多神经退行性疾病的细胞死亡中发挥作用。作为新生儿神经元细胞死亡的模型，我们研究了杂合性 Lurcher 突变体 (+/Lc) 中氧化应激在浦肯野细胞死亡中的作用。 Lurcher 是 82 谷氨酸受体 (GluR delta 2) 的功能获得性突变，该突变将受体转变为渗漏膜通道，导致 +/Lc 浦肯野细胞从出生后发育的第一周左右开始慢性去极化。事实上，所有 +/Lc 浦肯野细胞都会在出生后第一个月末死亡。为了研究氧化应激在 +/Lc 浦肯野细胞死亡中的作用，我们检测了野生型和 +/Lc 浦肯野细胞 P10、P15 和 P25 中一氧化氮合酶 (NOS) 活性以及两种氧化应激标记物硝基酪氨酸和锰超氧化物歧化酶 (MnSOD) 的表达。结果表明，NOS 活性以及硝基酪氨酸和 MnSOD 的免疫标记在 +/Lc 中增加。浦肯野细胞。为了确定过氧亚硝酸盐的形成是否是 +/Lc 浦肯野细胞死亡的先决条件，将 +/Lc 突变体与 α-nNOS 敲除突变体 (nNOS α(-/-)) 杂交以减少 NO 的产生。对双突变体的分析表明，阻断 α-nNOS 表达并不能拯救 +/Lc 浦肯野细胞。然而，我们提供了 GluR δ 中持续的 NOS 活性和硝基酪氨酸形成的证据。 2(+/Lc):nNOS(-/-) 双突变浦肯野细胞，这表明未能拯救 GluR delta 2(+/Lc) nNOS(-/-) 浦肯野细胞可能是由于诱导了替代的 nNOS 亚型 (c) 2007 Wiley periodicals, Inc.。
Oxidative stress is postulated to play a role in cell death in many neurodegenerative diseases. As a model of neonatal neuronal cell death, we have examined the role of oxidative stress in Purkinje cell death in the heterozygous Lurcher mutant (+/Lc). Lurcher is a gain of function mutation in the 82 glutamate receptor (GluR delta 2) that turns the receptor into a leaky membrane channel, resulting in chronic depolarization of +/Lc Purkinje cells starting around the first week of postnatal development. Virtually, all +/Lc Purkinje cells die by the end of the first postnatal month. To investigate the role of oxidative stress in +/Lc Purkinje cell death, we have examined nitric oxide synthase (NOS) activity and the expression of two markers for oxidative stress, nitrotyrosine and manganese super oxide dismutase (MnSOD), in wild type and +/Lc Purkinje cells at P10, P15, and P25.The results show that NOS activity and immunolabeling for nitrotyrosine and MnSOD are increased in +/Lc Purkinje cells. To determine whether peroxynitrite formation is a prerequisite for +/Lc Purkinje cell death, +/Lc mutants were crossed with an alpha-nNOS knockout mutant (nNOS alpha((-/-)) to reduce the production of NO. Analysis of the double mutants showed that blocking alpha-nNOS expression does not rescue +/Lc Purkinje cells. However, we present evidence for sustained NOS activity and nitrotyrosine formation in the GluR delta 2(+/Lc):nNOS(-/-) double mutant Purkinje cells, which suggests that the failure to rescue GluR delta 2(+/Lc) nNOS(-/-) Purkinje cells may be explained by the induction of alternative nNOS isoforms. (c) 2007 Wiley Periodicals, Inc.