MOLECULAR BASIS OF STRIATAL D1 SUPERSENSITIVITY

纹状体 D1 超敏性的分子基础

• 批准号: 2259895
• 负责人: DOUGLAS GENE COLE
• 金额: $ 7.66万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-07-01 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2259895
• 关键词: Parkinson's disease; antisense nucleic acid; binding proteins; biological signal transduction; calcium channel blockers; calcium flux; calmodulin dependent protein kinase; cyclic AMP; dopamine receptor; gene expression; immunocytochemistry; in situ hybridization; laboratory rat; messenger RNA; neuropharmacology; northern blottings; oligonucleotides; phosphorylation; receptor sensitivity; stereotaxic techniques; substantia nigra; tissue /cell culture

Dopamine receptor supersensitivity may underlie the dyskinesias that limit levodopa treatment for Parkinson's disease (PD). Its mechanism is unknown. Following destruction of the substantia nigra in human and rat, D1 receptor supersensitivity develops despite unchanged receptor number and affinity. our preliminary data indicate that nigral lesions alter D1- mediated regulation of cAMP- or Ca++-regulated signal transduction pathways, or both, suggesting the hypothesis that altered Intracellular signal transduction underlies D1 receptor supersensitivity. This application proposes to test this hypothesis. Aim 1 will determine whether D1 supersensitivity following nigral destruction is associated with altered expression of molecules in the cAMP signal transduction pathway, using Northern analyses and in situ hybridization. Aim 2 will determine if Ca++-regulated signal transduction pathways could be important for D1 supersensitivity by examining the effects of blocking different steps in Ca++-regulated pathways on D1- mediated CREB phosphorylation and Fos expression in primary striatal cell culture. Aim 3 will determine whether antisense phosphorothioate oligonucleotides can specifically inactivate signal transduction molecules important for D1 function in primary striatal cell culture. Aim 4 will test the hypothesis that upregulation of factors identified in earlier experiments plays a causal role in D1 supersensitivity by attempting to block rotational behavior in 6-OHDA-lesioned rats with intracerebral infusions of antisense phosphorothioate oligonucleotides. Successful completion of the proposed experiments will provide insight into the mechanisms of striatal adaptation to dopaminergic denervation and, in particular, D1 supersensitivity. The results may suggest new avenues for prevention or treatment of levodopa-induced dyskinesias in PD.

多巴胺受体超敏感性可能是限制运动的运动障碍的基础 左旋多巴治疗帕金森病（PD）。其机制尚不清楚。 人和大鼠的黑质遭到破坏后，D1 尽管受体数量不变，但受体超敏感性仍会发展 亲和力。我们的初步数据表明黑质病变改变了 D1- cAMP 或 Ca++ 调节信号转导的介导调节 途径，或两者，表明改变细胞内的假设 信号转导是 D1 受体超敏性的基础。这 应用程序建议检验这一假设。 目标 1 将确定黑质后 D1 是否超敏 破坏与 cAMP 中分子表达的改变有关 信号转导途径，使用 Northern 分析和原位 杂交。目标 2 将确定 Ca++ 是否调节信号转导 通过检查 D1 超敏感性途径可能很重要 阻断 Ca++ 调节途径中不同步骤对 D1- 的影响 原代纹状体细胞中介导的 CREB ​​磷酸化和 Fos 表达 文化。目标3将确定反义硫代磷酸酯是否 寡核苷酸可以特异性地灭活信号转导分子 对于原代纹状体细胞培养中的 D1 功能很重要。目标4将 检验早期发现的因子上调的假设 实验通过尝试在 D1 超敏感性中发挥因果作用 脑内阻断 6-OHDA 损伤大鼠的旋转行为 反义硫代磷酸寡核苷酸的输注。 成功完成拟议的实验将提供见解 纹状体适应多巴胺能去神经的机制 特别是 D1 超敏感性。结果可能会提出新的建议 预防或治疗 PD 中左旋多巴引起的运动障碍的途径。