Synaptic & intrinsic motoneuron excitability in ALS excitotoxicity

突触

• 批准号: 7777255
• 负责人: Jenna E Koschnitzky
• 金额: $ 2.83万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7777255
• 关键词: Adolescent; Adult; Alanine; Amino Acids; Amyotrophic Lateral Sclerosis; Animal Model; Bathing; Birth; Buffers; Calcium; Cells; Cessation of life; Characteristics; Chronic; Development; Disease; Effectiveness; Embryo; FDA approved; Familial Amyotrophic Lateral Sclerosis; Future; Genes; Glutamates; Glycine; Human; In Vitro; Ion Channel; Lead; Life; Maintenance; Mediating; Modeling; Motor Neurons; Mus; Mutate; Mutation; Napping; Neonatal; Neurodegenerative Disorders; Neurons; Paralysed; Patients; Pharmaceutical Preparations; Play; Preparation; Property; Relative (related person); Riluzole; Role; Route; Serum; Slice; Sodium; Spinal Cord; Superoxide Dismutase; Symptoms; Synapses; Therapeutic; Time; Up-Regulation; drug development; excitotoxicity; gamma-Aminobutyric Acid; mouse model; novel; patch clamp; presynaptic

DESCRIPTION (provided by applicant): Amyotrophic lateral sclerosis (ALS) is an adult onset neurodegenerative disease which specifically targets motoneurons. In 20% of familial ALS (fALS) cases, there is a specific mutation in superoxide dismutase (SOD1). Mice with mutated human SOD1, display the same pathological and phenotypical symptoms in human ALS patients and are therefore widely used as an animal model of fALS. However, the role SOD1 plays in determining motoneuron death remains unclear. A leading hypothesis is that excitotoxicity caused by an overload of calcium into the cell due to motoneuron hyperexcitability causes selective motoneuron death because motoneurons have a low capacity to buffer calcium. This proposal aims to determine the relative contribution of changes in intrinsic and synaptic motoneuron excitability between wild type and SOD1 motoneurons. Furthermore, the effect of Riluzole, the only FDA approved drug for ALS, on both intrinsic and synaptic motoneuron excitability will also be assessed. The aims of this proposal will be accomplished through the use of embryonic cultured motoneurons, neonatal spinal cord slices, and a new in vitro sacral cord preparation. The use of these preparations will allow an efficient and complete characterization of motoneuron excitability between wild type and SOD1 mice. The in vitro sacral cord preparation allows intrinsic and synaptic motoneuron excitability to be assessed through intracellular recordings at time points throughout development and adulthood. It also enables us to administer Riluzole at therapeutic levels and in multiple time courses to extensively characterize the effects of Riluzole over time. The neonatal spinal cord slice preparation allows intrinsic versus synaptic motoneuron excitability to be assessed using direct bath application of drugs to inhibit specific ion channels using a whole cell patch clamp configuration. Also, the direct and immediate effects of Riluzole on motoneurons can be determined. The embryonic cultured motoneurons provide a window into motoneuron abnormalities present before birth and also give us a preparation where no synaptic inputs are present. In this preparation the effects of Riluzole administered over multiple time courses on the intrinsic properties of motoneurons can be isolated. The cause(s) of motoneuron degeneration in ALS needs to be determined in order to develop novel drugs treatments and understanding differences or changes in motoneuron function will lead to specific targets for these future drugs. Furthermore, early detectable changes in motoneuron function may enable the use of pre-emptive therapies that can significantly delay or even arrest the development of the disease.

描述（由申请人提供）：肌萎缩性侧索硬化症（ALS）是一种成人发作神经退行性疾病，专门针对运动神经元。在20％的家族性ALS（FALS）病例中，超氧化物歧化酶（SOD1）存在特定的突变。突变的人类SOD1的小鼠在人ALS患者中表现出相同的病理和表型症状，因此被广泛用作伪造的动物模型。但是，SOD1在确定运动神经元死亡中所起的作用尚不清楚。一个主要的假设是，由于运动神经元的过度兴奋性导致选择性的运动神经元死亡，因此由于运动神经元的过度性而导致的钙毒性导致细胞，因为运动神经元具有较低的缓冲钙的能力。该建议旨在确定野生型和SOD1运动神经元之间内在和突触运动神经元兴奋性变化的相对贡献。此外，还将评估Riluzole（唯一获得FDA批准的ALS药物）对固有和突触运动神经元兴奋性的影响。该提案的目的将通过使用胚胎培养的运动神经元，新生儿脊髓切片和新的体外s骨绳制剂来实现。这些制剂的使用将使野生型​​和SOD1小鼠之间的运动神经元兴奋性有效，完整地表征。体外s骨绳的制备允许通过在整个发育和成年期的时间点上的细胞内记录来评估固有和突触的运动神经元兴奋性。它还使我们能够在治疗水平和多个时间课程中管理Riluzole，从而广泛地表征了Riluzole随时间的影响。新生儿脊髓切片的制剂允许使用药物直接浸泡药物使用全细胞斑块夹构型抑制特定的离子通道来评估固有的与突触运动神经元的兴奋性。同样，可以确定riluzole对运动神经元的直接和直接影响。胚胎培养的运动神经元在出生前提供了一个窗口，以探索运动神经元异常，还为我们提供了不存在突触输入的准备。在此准备中，可以分离出多个时间疗程对运动神经元内在特性的riluzole的影响。为了开发新的药物治疗并理解差异或运动神经元功能的变化，需要确定ALS中运动神经元变性的原因，这将导致这些未来药物的特定靶标。此外，运动神经元功能的早期可检测到的变化可能会实现可显着延迟甚至阻止疾病发展的先发制体疗法。