Formation and toxicity of peripherin inclusions

描述(由申请人提供)： 发动机中中间丝异常夹杂物的存在 神经元是肌萎缩侧索硬化症的共同病理特征 硬化症(ALS)。这些包涵体中的大多数由 神经丝(NF)蛋白和外周蛋白，正常情况下为III型 在运动神经元中低水平表达。最近，我们发现 野生型外周蛋白在小鼠体内的过表达引发了这种疾病的形成 IF包涵体与运动神经元迟发性死亡的关系。此外， 疾病是由核因子-光(NF-L)蛋白水平缺乏所致， 与肌萎缩侧索硬化症有关的现象。这里提出了一些实验，以 进一步确定外周蛋白异常是否可能导致ALS 发病机制。我们将产生一种新的转基因小鼠模型， 外周蛋白包裹体的形成，受多西环素调控 转基因表达。我们将研究形成和发展的调控机制。 神经细胞外周蛋白聚集体的毒性。以前的结果表明， 促炎细胞因子和细胞因子上调外周蛋白表达 兴奋性中毒性损伤。建议使用培养的细胞和 转基因小鼠的方法，以进一步定义调控元件 激活外周蛋白基因转录，并确定是否诱导 外周血外周蛋白水平与脑缺血后神经元丢失有关 兴奋性中毒性损伤。此外，基因敲除方法将使我们能够 确定外周蛋白在两个已建立的疾病发病机制中的作用 运动神经元病的小鼠模型，表达突变超氧化物的小鼠 与肌萎缩侧索硬化症和携带摇摆器突变的小鼠有关的歧化酶。最后，我们 将在家族性和散发性红斑狼疮病例中寻找外周蛋白基因突变 肌萎缩侧索硬化。

DESCRIPTION (provided by applicant): The presence of abnormal inclusions of intermediate filaments (IFS) in motor neurons represents a common pathological feature of amyotrophic lateral sclerosis (ALS). The majority of these inclusion bodies are composed of neurofilament (NF) proteins together with peripherin, a type III IF normally expressed at low levels in motor neurons. Recently, we discovered that the overexpression of wild-type peripherin proteins in mice provokes the formation of IF inclusion bodies and late-onset death of motor neurons. Moreover, the disease was precipitated by a deficiency in levels of NF light (NF-L) proteins, a phenomenon associated with ALS. A number of experiments are proposed here to further determine whether peripherin abnormalities may contribute to ALS pathogenesis. We will generate a new transgenic mouse model with the onset of peripherin inclusion formation, modulated by the doxycycline control of transgene expression. We will study the mechanisms regulating the formation and toxicity of peripherin aggregates in neurons. Previous results demonstrated an upregulation of peripherin expression by pro-inflammatory cytokines and by excitotoxic injury. It is proposed, with the use of cultured cells and of transgenic mouse approaches, to further define the regulatory elements activating peripherin gene transcription and to determine whether induction of peripherin levels contributes to neuronal loss after cerebral ischemia and excitotoxic injury. In addition, the gene knockout approach will allow us to determine the contribution of peripherin to pathogenesis in two established mouse models of motor neuron disease, mice expressing mutant superoxide dismutase linked to ALS and mice carrying the wobbler mutation. Finally, we will search for peripherin gene mutations in familial and sporadic cases of ALS.