Non-invasive Chemical Genetic Control of Neuronal Activity

神经元活动的非侵入性化学遗传控制

• 批准号: 7684412
• 负责人: MICHAEL D EHLERS
• 金额: $ 39万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7684412
• 关键词: Action Potentials; Acute; Afferent Neurons; Agonist; Animals; Area; Autistic Disorder; Behavior; Behavioral; Blood - brain barrier anatomy; Brain; Cannulas; Capsaicin; Cells; Charge; Data; Development; Dopamine; Dose; Engineering; Epilepsy; FOS gene; Fluorescent Dyes; Gated Ion Channel; Genetic; Glutamates; Goals; Implant; Infusion procedures; Injection of therapeutic agent; Knock-in Mouse; Laboratories; Lidocaine; Ligands; Maps; Membrane; Memory; Mental disorders; Metabolic; Metabolism; Methods; Microdialysis; Modeling; Monitor; Mus; Neurologic; Neurons; Neurosciences; Nociception; Operative Surgical Procedures; Optics; Pan Genus; Peripheral; Pharmaceutical Preparations; Physiological; Population; Public Health; Purkinje Cells; Reading; Regulation; Research; Resiniferatoxin; Response Latencies; Route; Schizophrenia; Slice; Sodium Channel; Sodium Channel Blockers; Specificity; Staining method; Stains; Stereotyped Behavior; System; TRPV1 gene; Techniques; Technology; Testing; Therapeutic; Time; Work; addiction; awake; base; behavior influence; brain cell; cell type; chemical genetics; depression; direct application; dopaminergic neuron; implantation; in vivo; interest; light gated; mouse model; neural circuit; neural stimulation; new technology; novel; programs; public health relevance; public health research; receptor; relating to nervous system; research study; response; sensory stimulus; small molecule; technology development

DESCRIPTION (provided by applicant): Mapping functional circuits is a major goal for both cellular and systems neuroscience. Current approaches for mapping neural circuits are limited by the lack of technologies for evoking cell-specific neural activity. Available methods of neural stimulation rely on either local application of undiscriminating fields of electrical currents, glutamate uncaging, or the presentation of artificial sensory stimuli. Although recent use of light- gated ion channels has provided optical control of neuronal activity on rapid time scales, such approaches are limited by the requirement for direct optical access to neuronal populations of interest, and are not currently suitable for activating large brain areas or disperse neuronal populations. A transformative technology for neuroscience would be non-invasive control over neural activity in genetically defined populations of neurons in the mammalian brain. Such a goal requires combining genetic sensitization of neuronal subsets with a means to manipulate their electrical activity remotely without surgery or intracranial implants. To create such a technology, my laboratory has initiated a program of in vivo chemical genetic and physiological studies to engineer a mouse model suitable for precise non-invasive manipulation of neural activity in genetically defined populations of neurons in vivo. We have developed a conditional mouse model that sensitizes genetically defined neurons to an artificial ligand (capsaicin) by cell type-specific expression of a heterologous receptor (TRPV1). We have found that application of capsaicin to neurons expressing TRPV1 induces strong inward currents, triggers robust firing of action potentials, and activates stereotyped behaviors. Taking advantage of these preliminary data, and the extensive pharmacological and biophysical characterization of TRPV1, we propose to extend and modify this model to enable peripheral administration of agonists for central activation of defined neuronal subsets. Moreover, because the large TRPV1 channel pore is permeable to small molecules, including the membrane-impermeant sodium channel blocker QX-314, we propose to test this novel mouse model to enable both activation and inhibition of neuronal activity. This work will allow for the development of a novel in vivo technology for chemical genetic regulation of neuronal activity that is (1) orthogonal to optical and optogenetic strategies, (2) based on the only current Cre/lox-based model for neuronal activation, (3) may allow for fully non-invasive CNS activation by drug injection, and (4) may enable targeted small molecule delivery to defined neuronal subsets. PUBLIC HEALTH RELEVANCE: The proposed research will develop a novel technology for non-invasive control over electrical activity in genetically defined populations of brain cells. Abnormal electrical activity in the brain contributes to epilepsy, memory decline, depression, autism, schizophrenia, and addiction. By developing a crucial new technology for targeted manipulation of brain cell activity and metabolism, the proposed research will define novel brain circuits and therapeutic strategies for treating these devastating neurological and psychiatric disorders, which currently have a profound negative impact on public health.

描述（由申请人提供）：映射功能电路是细胞和系统神经科学的主要目标。当前的映射神经回路方法受到唤起细胞特异性神经活动的技术的限制。可用的神经刺激方法依赖于局部应用电流，谷氨酸渗透的不歧视场或人造感觉刺激的呈现。尽管最近使用的光离子通道已在快速时间尺度上提供了神经元活动的光学控制，但这种方法受到直接光学访问感兴趣的神经元群体的要求限制，目前不适合激活大脑区域或分散神经元种群。神经科学的变革性技术将是对哺乳动物大脑中神经元种群中神经活动的非侵入性控制。这样的目标需要将神经元亚群的遗传敏化与无需手术或颅内植入物操纵其电活动的手段。为了创建这样的技术，我的实验室启动了一个体内化学遗传和生理研究程序，以设计一种小鼠模型，适用于体内神经元遗传定义的神经活动的精确非侵入性操纵。我们开发了一种条件小鼠模型，该模型通过异源受体（TRPV1）的细胞类型特异性表达将遗传学定义的神经元敏感到人工配体（辣椒素）。我们发现，辣椒素在表达TRPV1的神经元中的应用会诱导强大的内向电流，触发动作电位的强大发射并激活陈规定型行为。利用这些初步数据以及TRPV1的广泛药理和生物物理表征，我们建议扩展和修改该模型，以使激动剂的外围给药以中心激活定义的神经元亚群。此外，由于较大的TRPV1通道孔可渗透到小分子中，包括膜 - 覆盖钠通道阻滞剂QX-314，我们建议测试这种新型小鼠模型以启用神经元活性的激活和抑制。这项工作将允许开发一种新型的体内技术，用于（（1）基于唯一基于CRE/LOX的神经元激活的CRE/LOX模型，（2）神经元活动的化学遗传调节（（1）（1），（2）可以通过药物注射和较小的分子来启动非恒星CNS激活（4）。公共卫生相关性：拟议的研究将开发一种新的技术，用于在遗传定义的脑细胞种群中非侵入性控制电活动。大脑的异常电活动有助于癫痫，记忆力下降，抑郁，自闭症，精神分裂症和成瘾。通过开发针对脑细胞活性和代谢的针对性操纵的至关重要的新技术，拟议的研究将定义新的脑回路和治疗策略，以治疗这些毁灭性的神经系统和精神疾病，目前对公共卫生产生了深远的负面影响。