Optical control of neuromodulatory GPCRs

总结/摘要 神经科学的一个主要目标是了解神经调节系统如何调节大脑的核心过程 和行为，从运动功能和学习到奖励，厌恶，注意力和睡眠。这些系统 在精神分裂症和情绪、运动控制和认知障碍中出现偏差。这些疾病的治疗通常 转向神经调质及其受体的药理学操作。了解 由于光遗传学和化学遗传学，神经调节回路已经有了相当大的进步。但 神经调节很难破解一个主要的障碍是一个单一的神经调质可能发挥多种多样的作用， 因为它有多种受体，在不同的细胞位置和不同的细胞中具有不同的功能， 在一个电路中。不幸的是，药物和基因操作通常不能被靶向或控制， 足够的时空精度来解开这些不同的功能。我们正在开发两种方法 通过控制天然的全长神经调节受体来提供所需的精确度， 光可转换的栓系配体（PTL）。初步工作证明了这两种方法的可行性， 应用于两个核心神经调节受体家族：代谢型谷氨酸受体（mGluRs）和 多巴胺受体（DARs）。在本提案中，我们优化并扩展了一种直接 将PTL连接到特定的受体亚型上，并开发出一种不接触这些受体的新方法， 而是在膜锚上将PTL递送给它们。因为膜锚可以靶向特定的 在细胞内的位置，它可以选择性地控制受体只在该位置，它可以使用或广泛的 配体（其击中该类型的所有局部受体）或高选择性配体（以在一个时间点上仅控制一种亚型）。 时间）。这些工具将在小鼠中进行演示，并作为基因传递载体（PTL）提供给社区。 和转基因动物品系。

SUMMARY/ABSTRACT A major goal of neuroscience is to understand how neuromodulatory systems regulate core processes of brain and behavior, from motor function and learning to reward, aversion, attention, and sleep. These systems go awry in schizophrenia and disorders of mood, motor control and cognition. Treatment for these conditions often turns to pharmacological manipulation of neuromodulators and their receptors. Understanding of neuromodulatory circuits has advanced considerably thanks to optogenetics and chemogenetics. But neuromodulation is difficult to crack. A major obstacle is that a single neuromodulator may play many diverse roles because it has multiple receptors, with different functions in different cell locations and different cells within a circuit. Unfortunately, drugs and genetic manipulations cannot typically be targeted or controlled with sufficient spatio-temporal precision to unravel these different functions. We are developing two approaches that provide the needed precision by controlling native, full-length neuromodulatory receptors with photoswitchable tethered ligands (PTLs). Preliminary Iwork demonstrates feasibility of both approaches as applied to two core neuromodulatory receptor families: metabotropic glutamate receptors (mGluRs) and dopamine receptors (DARs). In this proposal, we optimize and expands an approach in which we directly attach PTLs to a specific receptor subtype and develop a new approach that does not touch those receptors, but deliver the PTL to them on a membrane anchor. Because the membrane anchor can be targeted to specific locations within the cell, it can selectively control receptors only at that location, and it can use either broad ligands (which hit all local receptors of that type) or highly selective ligands (to control only one subtype at a time). The tools will be demonstrated in mice and provided to the community as gene delivery vectors, PTLs and transgenic animal lines.