SPINAL CORD CELL EXCITABILITY

Wobbler小鼠是一种研究常染色体隐性遗传的动物模型 运动神经元病(MND)。这些动物中约25%会患上MND相关疾病 对颈髓细胞的病理学影响。可能导致 导致颈髓运动神经元功能障碍和死亡，如肌萎缩症 侧索硬化症(ALS)，目前还没有完全了解。的目标是 这项研究是对神经递质异常的识别- 在存活的脊髓细胞群体中的受体功能。在工作中 这个项目的假设是摇摆人中的基因异常 小鼠可能通过兴奋性氨基酸的变化来表达表型 颈髓细胞的酸受体功能。缺陷可能是 兴奋性反应中膜电位异常的反映 氨基酸。这一假设的基本原理是基于这样一个角色 兴奋性氨基酸及有关物质，如P-N-甲基氨基-L- 丙氨酸(BMAA)在运动神经元细胞毒性中发挥作用。电气和化工 将研究摇摆器中存活的脊髓细胞的兴奋性 小鼠采用流式细胞术和电压敏感型染料。相对膜 兴奋性氨基酸、电压依赖性神经毒素的潜在变化， 抑制性氨基酸和神经肽将在前两部分进行分析， 分别是第三、第四和第五年。摇摆器(NFR/WR)和控制 (NFR)脊髓将被分6个区域解剖，细胞将被 使用木瓜酶解离方案进行分离。在审查之后 生活力、相对膜电势的变化会对处理 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 新墨西哥大学和洛斯阿拉莫斯国家实验室。这个 策略将允许对各种不同的兴奋性进行纵向评估 各区域以及不同的出生前和出生后年龄，包括之前的年龄 临床症状的发展。心理应激性的研究 正常同源基因株(NFR)将控制实验结果。 使用电压敏感的单细胞定量荧光显微镜 染料将补充流式细胞仪的种群数据。这个 对可能的膜电位异常的反应特征 Wobbler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.