Acrylamide and Diketone Neurotoxicity: Axonal transport

丙烯酰胺和二酮神经毒性：轴突运输

• 批准号: 6546424
• 负责人: DALE William SICKLES
• 金额: $ 21.53万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6546424
• 关键词: Drosophilidae; acrylamides; antisense nucleic acid; axon; cell membrane; electrophysiology; environmental toxicology; gene expression; immunofluorescence technique; immunoprecipitation; ketones; kinesin; molecular pathology; motor neurons; neural degeneration; neural transmission; neuromuscular junction; neuronal transport; neuropsychology; neurotoxicology; occupational hazard; plastics; polymers; synapses; tissue /cell culture; video microscopy; western blottings

DESCRIPTION (provided by applicant): Extensive production of acrylamide and gamma-diketones for diverse industrial, agricultural and commercial use creates the potential for occupational and environmental neurotoxicity. Additionally, these agents act as prototypes for other toxicants as well as models for neuropathologies. While research has identified characteristics of the neurotoxicity, pathogenetic mechanisms remain unresolved. This proposal logically extends our previous studies by evaluating inhibition of the motor protein kinesin, compromise of fast anterograde axonal transport (faAXT) and reduced delivery of distal axonal/terminal proteins with subsequent development of dysfunctional axons and/or terminals as a significant contributant to behavioral deficits (neurotoxicity). 1) The significance of faAXT reductions will be tested by challenging the association of reduced transport with neurotoxicity over a range of dosing rates using video-enhanced DIC microscopy of isolated axons. 2) Changes in quantity of fast-transported proteins, specific for axolemmal, synaptic vesicle and synaptic plasma membrane compartments and possessing neurophysiological relevance, will be determined with immunofluorescence, Western blots and/or immunoprecipitation. Different dosing rate paradigms as well as quantitation in PNS and CNS axons and terminals are planned. 3) Electrophysiological testing of axonal conduction and neurotransmission efficacy will challenge the association of protein deficits with functional outcomes and may identify specific molecular mechanisms related to behavioral deficits. 4) Specific and quantitative alterations in neuronal kinesin content using antisense and overexpression techniques are integrated with determination of sensitivity of faAXT, morphological changes and/or distal axonal protein content to toxicant challenges. The integrative design permits quantitative comparison of key elements of the hypothesized pathogenic cascade while simultaneously challenging the relationship of these outcomes to neurotoxicity. These studies will improve our understanding of compromised fast axonal transport with these toxicants and in neurodegenerative disorders, in general. These studies will enhance the evaluation of the emerging concept of motor protein defects producing specific peripheral neuropathies.

描述（由申请人提供）：用于多种工业，农业和商业用途的丙烯酰胺和γ-二酮的广泛生产为职业和环境神经毒性带来了潜力。此外，这些药物是其他有毒物质的原型，以及神经病理学的模型。尽管研究已经确定了神经毒性的特征，但致病机制仍未解决。从逻辑上讲，该提案通过评估抑制运动蛋白动力素，快速行进轴突运输（FAAXT）的损害以及减少远端轴突/末端蛋白的递送，并随后发展功能障碍轴突和/或末端的重要性，从而扩大了我们先前的研究。缺陷（神经毒性）。 1）通过使用分离的轴突的视频增强DIC显微镜在一系列给药率的范围内，将通过挑战降低运输与神经毒性的关联来测试FAAXT降低的重要性。 2）将通过免疫荧光，蛋白质印迹和/或免疫致敬来确定，将针对轴突，突触囊泡和突触质膜室的快速传输蛋白质变化，并具有神经生理学相关性。计划在PNS和CNS轴突和终端中进行不同的给药率范例以及定量。 3）轴突传导和神经传递功效的电生理测试将挑战蛋白质缺陷与功能结果的关联，并可以识别与行为缺陷有关的特定分子机制。 4）使用反义和过表达技术在神经元驱动蛋白含量中的特定和定量改变，并确定FAAXT，形态变化和/或轴突蛋白蛋白含量含量对有毒挑战的敏感性。综合设计允许对假设的致病级联反应的关键要素进行定量比较，同时挑战了这些结果与神经毒性的关系。这些研究将一般来说，这些研究将提高我们对这些有毒物质和神经退行性疾病的快速轴突运输的理解。这些研究将增强对产生特定周围神经病的运动蛋白缺陷的新兴概念的评估。