Selective modulation of basal ganglia circuits

基底神经节回路的选择性调制

• 批准号: 6625940
• 负责人: SCOTT J SHERMAN
• 金额: $ 18.94万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-15 至 2004-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6625940
• 关键词: Adenoviridae; Herpesviridae; antisense nucleic acid; basal ganglia; gamma aminobutyrate; gene expression; green fluorescent proteins; immunocytochemistry; laboratory rat; nerve threshold; neurogenetics; potassium channel; protein structure function; putamen; recombinant proteins; transfection; transfection /expression vector; voltage /patch clamp; voltage gated channel

Parkinson's Disease (PD) is a neuro-degenerative disorder of the basal ganglia in which there is a loss of dopamine-containing neurons in the substantia nigra, which project to the putamen and caudate (striatum). When the loss of dopaminergic innervation in the striatum reaches a critical level, motor symptoms such as tremor, rigidity, and brady kinesia become manifest. The availability of animal models of PD have led to a heightened understanding of the basal circuitry, and underscored the importance of neuronal activity in two parallel striatal outflow pathways termed the "direct" and "indirect" pathway. The relative activity of the pathways is disturbed in the Parkinsonian state. This proposal explores novel techniques aimed at differentially controlling activity in these two pathways using gene transfer vectors that alter neuronal excitability. Viral vectors based on herpes virus or adenovirus will be constructed and used to augment the voltage-gated potassium channel activity in cultured striatal neurons from rat. Single-cell electrophysiology and immunocytochemistry will be used to monitor the effects of this channel modulation on defined cell types corresponding to the direct or indirect pathway. The specific aims of this proposal are: (1) to determine whether of a useful model system of basal ganglia circuitry can be developed using dispersed primary cultures of rat putaminal neurons; (2) to determine whether viral gene transfer vectors using cell-specific promoters are capable of selectively transducing cells belonging to the indirect or direct putaminal output pathways, and (3) to develop and test a gene transfer vector designed to selectively augment K+ channel activity in basal ganglia neurons belonging to either the direct or indirect output pathways. Successful completion of this proposal would allow further the development of a gene therapy for the symptomatic treatment of PD.

帕金森病 (PD) 是一种基底神经节神经退行性疾病，其中黑质中投射到壳核和尾状核（纹状体）的含多巴胺神经元缺失。当纹状体多巴胺能神经支配的丧失达到临界水平时，震颤、僵硬和运动缓慢等运动症状就会变得明显。 PD动物模型的可用性提高了对基底回路的了解，并强调了神经元活动在称为“直接”和“间接”途径的两条平行纹状体流出途径中的重要性。在帕金森病状态下，通路的相对活动受到干扰。该提案探索了旨在使用改变神经元兴奋性的基因转移载体差异控制这两条途径的活性的新技术。将构建基于疱疹病毒或腺病毒的病毒载体，并用于增强培养的大鼠纹状体神经元中的电压门控钾通道活性。单细胞电生理学和免疫细胞化学将用于监测该通道调节对与直接或间接途径相对应的特定细胞类型的影响。该提案的具体目标是：（1）确定是否可以使用大鼠壳核神经元的分散原代培养物开发有用的基底神经节电路模型系统； (2) 确定使用细胞特异性启动子的病毒基因转移载体是否能够选择性转导属于间接或直接壳核输出途径的细胞，以及 (3) 开发和测试设计用于选择性增强属于直接或间接输出途径的基底神经节神经元中 K+ 通道活性的基因转移载体。该提案的成功完成将有助于进一步开发用于帕金森症对症治疗的基因疗法。