Intact Circuit Assessment of Aging Dopamine Neurons vis Optogenetics and CLARITY

老化多巴胺神经元的完整电路评估与光遗传学和清晰度

• 批准号: 8712256
• 负责人: Viviana Gradinaru
• 金额: $ 37.46万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8712256
• 关键词: Accounting; Affect; Age; Aging; Alzheimer' s Disease; Anatomy; Area; Behavior; Behavioral; Brain; Brain Diseases; Brain Mapping; Cell Nucleus; Cell physiology; Cells; Cognition; Dependence; Development; Disease; Dopaminergic Cell; Electrophysiology (science); Frequencies; Genetic; Glutamates; Grouping; Heterogeneity; Immunohistochemistry; Individual; Laboratories; Life Expectancy; Light; Link; Literature; Location; Maps; Measures; Methods; Midbrain structure; Motor; Movement; Nerve Degeneration; Neuroanatomy; Neurobiology; Neurons; Neurosciences; Nutritional; Opsin; Parkinson Disease; Pathway interactions; Pattern; Phenotype; Positioning Attribute; Predisposition; Process; Property; Proteins; Quality of life; Rattus; Reporting; Research; Resistance; Rodent; Role; Signal Transduction; Slice; Specific qualifier value; Specificity; Speed; Structure of subthalamic nucleus; Substantia nigra structure; Synapses; System; Techniques; Testing; Time; Transgenic Organisms; Tyrosine 3-Monooxygenase; Ventral Tegmental Area; Work; age related; aged; aging brain; base; cell type; dopaminergic neuron; effective therapy; in vivo; innovation; microbial; millisecond; motor deficit; neural circuit; neuronal excitability; neurotransmission; normal aging; optogenetics; pars compacta; promoter; protein expression; public health relevance; relating to nervous system; research study; response; stem; targeted delivery; tool

DESCRIPTION (provided by applicant): During aging, motor function declines, with deficits in fine and fast movement and coordination. Experimental studies associate age-dependent motor deficits with the malfunction of dopaminergic (DA) pathway, which originates in the substantia nigra pars compacta (SNc). However we do not understand how the activity of DA neurons varies throughout aging in the different tiers of nigral neurons in vivo, what type of activity changes precede neurodegeneration, how these activity changes affect behavior, and whether restoring perturbed activity can delay neurodegeneration and/or behavioral deficits. To characterize, for the first time in the intact circuit, the function and anatomy of aging nigral dopaminergic circuits, we propose to use two powerful technological advances in neuroscience: one for cell-type specific bidirectional control of neuronal activity in vivo with high temporal precision (optogenetics); and one for intact brain circuit mapping and phenotyping, slicing-free (CLARITY). Optogenetics uses microbial opsins, light-sensitive proteins that can be expressed in specified cells via targeting promoters and turned on/off with millisecond speed, thus providing control of cell function with high spatial, temporal, and genetic specificity. Their abilty to control the electrical activity of neural circuits and confer reversible gain and loss of functin of specific neuronal phenotypes allows us to study neural systems and diseases in unprecedented manner. To target subsets of SNc DA neurons we will take advantage of the TH- Cre transgenic lines as well as localized stereotaxic opsin delivery and targeted light application We hypothesize that throughout aging, DA neurons in different SNc tiers have distinct behavioral contributions (Aim 1), which is due to differences in their intrinsic excitability (Aim ) and changes in synaptic inputs (Aim 3). This proposal combines powerful complementary techniques (optogenetics, electrophysiology, and neuroanatomy by CLARITY) to advance our understanding of dopaminergic function and contribution to behavior throughout aging by performing studies in the intact circuit. The PI has been involved in the development of both techniques and our laboratory is ideally positioned to apply these techniques to the aging brain with a focus on the DA system. A better understanding of the properties of DA neurons in the aging SNc can aid in identifying circuit targets and/or behavioral/nutritional methods to delay/reverse age-related alterations in these neurons and in motor functions.

描述（由申请人提供）：在老化期间，运动功能会降低，并且缺陷在罚款和快速运动和协调中。实验研究将依赖年龄的运动缺陷与多巴胺能（DA）途径的故障相关联，该途径起源于Nigra pars compacta（SNC）。但是，我们不了解DA神经元的活性在体内不同层次的不同层次的整个衰老中如何变化，哪种类型的活性在神经变性之前发生了变化，这些活动的变化如何影响行为以及恢复扰动的活性是否会延迟神经变性和/或行为赤字。为了表征完整电路中的第一次，衰老的nigral多巴胺能回路的功能和解剖结构，我们建议在神经科学中使用两种强大的技术进步：一种用于细胞型特异性双向对体内神经元活性的双向控制，具有高颞叶精度（optogenetics）；一个用于完整的脑电路映射和表型，无切片（清晰度）。光遗传学使用微生物蛋白，光敏蛋白可以通过靶向启动子在指定的细胞中表达并以毫秒速度打开/关闭，从而以高空间，时间和遗传特异性提供对细胞功能的控制。他们对控制神经回路的电活动的痛苦，并赋予特定神经元表型功能的可逆增益和功能丧失，使我们能够以前所未有的方式研究神经系统和疾病。为了靶向SNC DA神经元的子集，我们将利用Thcre cree转基因线以及局部的立体定位Opsin递送和靶向光应用，我们假设在整个衰老中，不同SNC层中的DA神经元具有独特的行为贡献（AIM 1），这是由于其内在兴奋性差异（AIMINIC兴奋性差异（AIM）（AIM）和AIMATS PASTICT（AIM PASTICT）（AIM PASTICT 3）。 该提案结合了强大的互补技术（通过清晰度的光遗传学，电生理学和神经解剖学），以提高我们对多巴胺能功能的理解，并通过在完整电路中进行整个衰老。 PI参与了这两种技术的开发，我们的实验室的理想位置可以将这些技术应用于衰老的大脑，重点是DA系统。更好地了解衰老SNC中DA神经元的性质可以帮助识别电路靶标和/或行为/营养方法，以延迟这些神经元和运动功能中与年龄相关的变化。