Two-photon optical control of astrocytic function

描述（由申请人提供）：星形胶质细胞不仅是回路的被动成员，而且它们也可能直接参与神经元计算。事实上，星形胶质细胞非常适合在回路中发挥潜在的广泛作用：它们以近乎完整的覆盖CNS，通过间隙连接连接成广泛的合胞体，并且来自单个星形胶质细胞的过程可以接触多达数万个突触。事实上，我们最近已经证明，即使是单个星形胶质细胞的刺激也可以控制皮层中神经元的回路同步。研究星形胶质细胞的工具比研究神经元功能的工具要少得多，因此，大多数关于星形胶质细胞信息处理的研究都是有限和不完整的。星形胶质细胞研究人员需要有选择地测量和操纵体内星形胶质细胞功能的方法。双光子激发，穿透活组织，同时提供单细胞分辨率，结合兼容的光化学或光遗传学探针，可能是未来星形胶质细胞研究的理想技术平台。我们建议开发新的双光子和计算工具，以提供亚细胞和电路水平的分析星形胶质细胞的功能在体内的神经回路，使用nvel星形胶质细胞特异性Brainbow小鼠线。这些工具包括双光子光遗传学和笼状化合物，在3D中成像星形胶质细胞群的方法，以及揭示星形胶质细胞与回路中神经元相互作用的计算算法。有了这个奖项，我们将提供 本发明提供了一种利用广泛的光学和计算工具来研究星形胶质细胞群落的方法，所述工具可以容易地适应于在体内使用。这项工作将使调查的作用，星形胶质细胞在塑造神经元活动的皮层微电路和超越，从而可能有一个根本性的影响，我们如何看待神经胶质细胞的处理，以及巩固的作用，一个被忽视的电路组成-星形胶质细胞-在皮层同步和计算。最后，这项工作可以引入一种新的潜在手段来检测或控制神经元功能，用于潜在的治疗目的：通过特异性激活星形胶质细胞。

DESCRIPTION (provided by applicant): Astrocytes are not merely passive members of the circuit, but that they may also be directly involved with neuronal computation. Indeed, astrocytes are well suited to a potential widespread role in the circuit: they tile the CNS with near- complet coverage, are connected into an extensive syncytium via gap junctions, and processes from a single astrocyte can contact up to tens of thousands of synapses. In fact, we have recently shown how even the stimulation of single astrocyte can control circuit-wide neuronal synchronizations in the cortex. Tools to study astrocytes are significantly less developed than those available to investigate neuronal function and, consequently, most research on astrocytic information processing has been limited and incomplete. Astrocyte researchers need methods to selectively measure and manipulate the function of astrocytes in vivo. Two-photon excitation, which penetrates living tissue while affording single-cell resolution, combined with compatible optochemical or optogenetic probes, could be an ideal technical platform for future astrocyte research. We propose to develop novel two-photon and computational tools to provide subcellular and circuit-level analysis of astrocytic function in neural circuits in vivo, using a nvel astrocyte-specific Brainbow mouse line. These tools include two- photon optogenetic and caged compounds, methods to image groups of astrocytes in 3D, and computational algorithms to reveal interactions of astrocytes with neurons in the circuit. With this award, we will provide the astroglia community with a wide range of optical and computational tools that can be readily adapted for use in vivo. This work will enable the investigation of the roles of astrocytes in shaping neuronal activity in the cortical microcircuit and beyond, and thus could have a fundamental impact on how we view neuro-glia processing, as well as on solidifying the role of an overlooked circuit constituent-the astrocyte-in cortical synchrony and computation. Finally, this work could introduce a novel potential means to assay or control neuronal function for potential therapeutic purposes: through specific activation of astrocytes.