SPINAL CORD CELL EXCITABILITY

Wobbler小鼠是一种常染色体隐性遗传动物模型，用于研究 运动神经元病（MND）。 这些动物中约有25%发展为MND相关 颈脊髓细胞的病理学。 可能导致 颈髓运动神经元功能障碍和死亡，如肌萎缩性 脊髓侧索硬化症（ALS）尚未完全了解。 的目标 这项研究是鉴定神经递质的异常， 受体功能的群体中活的脊髓细胞。 工作 这个项目假设是摇摆者的遗传异常 小鼠的表型可能通过兴奋性氨基酸的改变来表达， 颈髓细胞的酸受体功能。 缺陷可能是 反映在膜电位异常的兴奋性反应 个氨基酸 这一假设的基本原理是基于以下作用， 兴奋性氨基酸和相关物质，如P-N-甲基氨基-L- 丙氨酸（BMAA），在运动神经元细胞毒性中起作用。 电和化学 兴奋性将在来自Wobbler的活脊髓细胞中进行研究 小鼠使用流式细胞术和电压敏感染料。 相对膜 兴奋性氨基酸，电压依赖性神经毒素， 在前两种中分析抑制性氨基酸和神经肽， 第三、第四和第五年分别。 摇摆器（NFR/wr）和控制 (NFR)将脊髓分成6个区域， 使用木瓜蛋白酶解离方案分离。 专员查核 活力，相对膜电位变化的治疗将是 使用阴离子电压敏感染料oxonol和Coulter 753记录 双激光流式细胞仪 数据将使用以下计算机进行分析： 新墨西哥州大学和洛斯阿拉莫斯国家实验室 的 战略将允许在不同的兴奋性纵向评估 各地区和不同的产前和产后年龄，包括那些 临床症状的发展。 大脑皮层兴奋性的研究 正常同源株（NFR）对实验结果起控制作用。 单细胞定量荧光显微术使用电压敏感的 染料将补充流式细胞仪的人口数据。 的 表征响应于以下的可能膜电位异常： Wobbler小鼠脊髓细胞中的氨基酸神经递质可能 有助于了解Werdnig-Hoffman的发病机制 疾病和ALS。

The Wobbler mouse is an autosomal recessive animal model for the study of motor neuron disease (MND). About 25% of these animals develop MND related to pathology of cervical spinal cord cells. The mechanisms that might lead to cervical cord motor neuron dysfunction and death, as in amyotrophic lateral sclerosis (ALS), are yet to be completely understood. The goal of this study is the identification of abnormalities of neurotransmitter- receptor function in populations of viable spinal cord cells. The working hypothesis of this project is that genetic abnormalities in the Wobbler mouse may be phenotypically expressed by alterations in excitatory amino acid-receptor function of cervical spinal cord cells. The defects might be reflected by membrane potential abnormalities in response to excitatory amino acids. The rationale for this hypothesis is based on the role that excitatory amino acids and related substances, such as P-N-methyl amino-L- alanine (BMAA), play in motor neuron cytotoxicity. Electrical and chemical excitability will be studied in viable spinal cord cells from the Wobbler mouse using flow cytometry and voltage-sensitive dyes. Relative membrane potential changes to excitatory amino acids, voltage-dependent neurotoxins, inhibitory amino acids and neuropeptides will be analyzed in the first two, third, fourth and fifth years, respectively. Wobbler (NFR/wr) and control (NFR) spinal cords will be dissected in 6 regions and cells will be isolated using a papain dissociation protocol. After examination of viability, relative membrane potential changes to the treatments will be recorded using the anionic voltage-sensitive dye oxonol and a Coulter 753 dual laser flow cytometer. Data will be analyzed using the computers of the University of New Mexico and The Los Alamos National Laboratory. The strategy will permit a longitudinal assessment of excitability at various regions and at various pre and postnatal ages, including those that precede the development of clinical symptoms. The study of excitability in the normal congenic strain (NFR) will control for the experimental results. Single-cell quantitative fluorescence microscopy using voltage-sensitive dyes will complement the flow cytometer-population data. The characterization of possible membrane potential abnormality in response to amino acid neurotransmitters in Wobbler mouse spinal cord cells may contribute to the understanding of the pathogenesis of Werdnig-Hoffman disease and of ALS.