Sonogenetic control of neurons in a large volume of the rodent brain

摘要 神经科学中的一个关键挑战是开发非侵入性操作特定神经元的方法 活体内的细胞类型。虽然最近的光、化学和磁遗传方法已经彻底改变了我们的能力 为了同时控制神经细胞和非神经细胞类型，它们各自都存在严重的缺陷，包括 无法将光线投射到大脑深处的目标或大脑的大量区域(光敏)，以及缺乏 对化学和磁成因方法进行精确的时间控制。Chalasani实验室最近 演示了一种使用超声波控制神经元活动的非侵入性方法，他们称之为 声学遗传学。他们已经证明了线虫的机械敏感性Trp-N通道同源物， Hydractinia和Hydra Magipapillata可用于体外和体外无创激活哺乳动物细胞 在活体内。他们假设表达这些Trp-N通道的靶细胞对 非侵入性超声波产生的机械变形。这项建议旨在延长 通过声发生技术控制大量小鼠脑内特定神经元群体的研究 该系统将有助于扭转癫痫患者的电生理和行为缺陷， 举个例子。他们将通过测试变种和 Hydra Trp-N通道的嵌合体在高通量成像和切片培养电生理分析中的应用 在体外，以及在喂养和体内肌电分析(目标1)。他们还计划开发一种新的 基于铌酸锂的换能器，将把超声波传输到小鼠的整个大脑。具体来说，他们将 在将产生时空非相干超声的设备中使用施罗德的最佳漫射器设计 在反射时，避免了超声波中的干扰和局部尖峰(目标2)。最后，他们计划启动 GABA能抑制中间神经元广泛分布于大脑以减轻行为和电生理 癫痫和瑞德综合征小鼠模型的缺陷。光遗传学、化学遗传学和药理学 以前曾使用各种方法来控制这些细胞群，为比较提供了基准。 这些研究将开发一种非侵入性的方法来操纵大鼠体内特定细胞的活动 啮齿动物大脑或身体的体积。此外，这些方法可以被翻译到人类系统中来定向 用于治疗目的的特定细胞群。

Abstract A key challenge in neuroscience is the development of methods to non-invasively manipulate specific neuronal cell types in vivo. While recent opto-, chemo- and magneto-genetic approaches have revolutionized our ability to control both neuronal and non-neuronal cell types, they each suffer from critical drawbacks, including the inability to deliver light to targets deep within the brain or to large volumes of the brain (opto-), and the lack of precise temporal control for both chemo- and magneto-genetic approaches. The Chalasani lab has recently demonstrated a noninvasive method for controlling the activity of neurons using ultrasound, a system they call sonogenetics. They have demonstrated that mechanosensitive TRP-N channel homologs from C. elegans, Hydractinia, and Hydra magnipapillata can be used to non-invasively activate mammalian cells both in vitro and in vivo. They hypothesize that target cells expressing these TRP-N channels are rendered sensitive to mechanical deformations generated by non-invasive ultrasound waves. This proposal aims to extend the sonogenetic approach to control specific neuronal populations throughout large volumes of the mouse brain, a system that would be useful for reversing electrophysiological and behavioral deficits seen in epilepsy, for example. They will identify channels with non-overlapping ultrasound stimulus ranges by testing variants and chimeras of the Hydra TRP-N channels in high-throughput imaging and slice culture electrophysiology assays in vitro, as well as in feeding and electromyography assays in vivo (Aim 1). They also plan to develop a new lithium niobate-based transducer that will deliver ultrasound throughout the mouse brain. Specifically, they will use Schroeder’s optimal diffuser design in a device that will generate spatiotemporally incoherent ultrasound that upon reflection, avoids interference and localized spikes in ultrasound (Aim 2). Finally, they plan to activate GABAergic inhibitory interneurons broadly throughout the brain to alleviate behavioral and electrophysiological deficits in mouse models of epilepsy and Rhett’s syndrome. Optogenetic, chemogenetic, and pharmacological methods have been previously used to control these cell populations, providing benchmarks for comparison. These studies will develop a noninvasive method for manipulating the activity of specific cells within large volumes of the rodent brain or body. Further, these methods can be translated into the human system to target specific cell populations for therapeutic purposes.