Department of Bioengineering, Rice University;
Department of Bioengineering, Rice University.
J Vis Exp. 2020 Dec 22(166). doi: 10.3791/61352.
Acoustically Targeted Chemogenetics (ATAC) allows for the noninvasive control of specific neural circuits. ATAC achieves such control through a combination of focused ultrasound (FUS) induced blood-brain barrier opening (FUS-BBBO), gene delivery with adeno-associated viral (AAV) vectors, and activation of cellular signaling with engineered, chemogenetic, protein receptors and their cognate ligands. With ATAC, it is possible to transduce both large and small brain regions with millimeter precision using a single noninvasive ultrasound application. This transduction can later allow for a long-term, noninvasive, device-free neuromodulation in freely moving animals using a drug. Since FUS-BBBO, AAVs, and chemogenetics have been used in multiple animals, ATAC should also be scalable for the use in other animal species. This paper expands upon a previously published protocol and outlines how to optimize the gene delivery with FUS-BBBO to small brain regions with MRI-guidance but without a need for a complicated MRI-compatible FUS device. The protocol, also, describes the design of mouse targeting and restraint components that can be 3D-printed by any lab and can be easily modified for different species or custom equipment. To aid reproducibility, the protocol describes in detail how the microbubbles, AAVs, and venipuncture were used in ATAC development. Finally, an example data is shown to guide the preliminary investigations of studies utilizing ATAC.
声靶向化学遗传学（ATAC）允许对特定的神经回路进行非侵入性控制。ATAC 通过组合聚焦超声（FUS）诱导的血脑屏障开放（FUS-BBBO）、腺相关病毒（AAV）载体的基因传递以及工程化的、化学遗传学的、蛋白受体及其同源配体的细胞信号激活来实现这种控制。使用 ATAC，可以使用单次非侵入性超声应用以毫米精度转导大脑的大区域和小区域。这种转导以后可以在自由移动的动物中使用药物进行长期、非侵入性、无设备的神经调节。由于 FUS-BBBO、AAVs 和化学遗传学已在多种动物中使用，因此 ATAC 也应该可以扩展到其他动物物种的使用。本文扩展了以前发表的方案，并概述了如何在没有复杂的 MRI 兼容 FUS 设备的情况下，使用 MRI 引导优化 FUS-BBBO 对小大脑区域的基因传递。该方案还描述了鼠标靶向和约束组件的设计，这些组件可以由任何实验室 3D 打印，并且可以轻松修改为不同的物种或定制设备。为了提高可重复性，该方案详细描述了如何在 ATAC 开发中使用微泡、AAV 和静脉穿刺。最后，展示了一个示例数据，以指导使用 ATAC 的研究的初步调查。
