ACRYLAMIDE & HEXACARBON NEUROPATHY: THE MICROTUBULE

丙烯酰胺

• 批准号: 3253396
• 负责人: DALE William SICKLES
• 金额: $ 10.45万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-09-30 至 1992-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3253396
• 关键词: acrylamides; adenosinetriphosphatase; alkyl nitrile; crosslink; cytoskeleton; dosage; electron microscopy; environmental contamination; immunofluorescence technique; laboratory rat; microtubules; monoclonal antibody; neuronal transport; neurotoxins; occupational hazard; peripheral nervous system disorders; tissue /cell culture; tubulin; vinblastine

Peripheral neuropathy is a frequent consequence of exposure to toxic chemicals; neurofilamentous axonpathy is produced by exposure to acrylamide (ACR), hexacarbons (HXC), 3,3' iminodipropionitrile (IDPN). Determination of the doses and mechanism(s) of action of these compounds will allow accurate determination of permissable exposure limits, outline preventive measures, and possibly develop cures. This information is useful in the search for causes of other neuropathological conditions. We hypothesize that nerve degeneration observed in ACR and HXC neuropathies is due to block of fast axoplasmic transport, IDPN is used as a negative contorl. Since fast axoplasmic transport is dependent upon the microtubule, our focus is upon the effects of these toxicants upon this cytoskeletal element. We will determine the time frame and dose response of changes in axoplasmic transport following single injections of ACR,HXC and/or IDPN and determine whether these changes are caused by disassembly of microtubles in vivo and/or in vitro using immunofluorescence of cultured mouse Balb 3T3 cells with specific monoclonal antibodies, turbidity assays, biochemical analysis of polymerized vs non-polymerized tubulin and electron microscopy. Disruption of mitosis and axoplasmic transport in vivo and assembly of tubulin into microtubules in vitro by specific doses of the toxicants will be measured. Correlation of the altered axoplasmic transport rate and capacity with microtubule disruption will be conducted. We will determine whether or not these toxicants directly bind to the colchicine and/or vinblastine binding sites, whether they affect microsomal ATPase activity and whether tyey crosslink tubulin using western blots and specific monoclonal antibodies. The data gathered by these investigations will determine whether or not microtubule structural alteration or dissembly by the neurotoxicants is related to microtubule dysfunction and may determine the mechanism by which this action is mediated.

周围神经病是暴露的经常结果 有毒化学物质；神经丝轴突路径是通过暴露产生的 到丙烯酰胺（ACR），己缩（HXC），3,3'Iminodipropionitrile （IDPN）。 确定剂量和机制的作用 这些化合物将允许准确确定可允许的 暴露限制，概述预防措施，并可能发展 治疗。 此信息在搜索原因 其他神经病理条件。 我们假设在ACR和HXC中观察到神经变性 神经病是由于快速轴突运输的阻滞，IDPN为 用作负面的。 由于快速轴突运输是 取决于微管，我们的重点是 这些有毒物质在该细胞骨架元件上。 我们将确定 轴质转运变化的时间范围和剂量响应 在单次注射ACR，HXC和/或IDPN并确定 这些更改是否是由于微透明的拆卸而引起的 使用培养小鼠的免疫荧光体内和/或体外体外 BALB 3T3细胞具有特定的单克隆抗体，浊度 测定，聚合与非聚合的生化分析 微管蛋白和电子显微镜。 有丝分裂和 体内的轴质转运和小管蛋白组装到 特定剂量的有毒物质在体外的微管将是 测量。 轴质转运率改变的相关性和 将进行微管破坏的容量。 我们将 确定这些毒物是否直接与 秋水仙碱和/或vinblastine结合位点，无论它们是否影响 微粒体ATPase活性以及Tyey交叉链接小管是否使用 蛋白质印迹和特定的单克隆抗体。 这些调查收集的数据将确定是否 或者不是微管结构的改变或由 神经毒性剂与微管功能障碍有关，可能 确定该作用介导的机制。