用于狨猴大脑精确兴奋和抑制的特定投射交叉光遗传学

Projection-Specific Intersectional Optogenetics for Precise Excitation and Inhibition in the Marmoset Brain.

作者信息

Shaw Luke, Padmanabhan Krishnan, Buckleaw Amy, Mitchell Jude F, Wang Kuan Hong

机构信息

Department of Neuroscience, Del Monte Institute for Neuroscience, University of Rochester Medical Center, Rochester, NY 14642, USA.

Department of Brain and Cognitive Sciences, University of Rochester, Rochester, NY 14611, USA.

出版信息

bioRxiv. 2025 Jun 22:2025.06.18.660378. doi: 10.1101/2025.06.18.660378.

Abstract

The primate cerebral cortex relies on long-range connections to integrate information between functionally specialized areas. Investigating these processes requires tools that can selectively modulate specific projection pathways. While cell-class-specific optogenetics can modulate local circuits, these approaches often lack pathway specificity. Projection-specific optogenetics offers greater precision, especially in primates, where cortical areas are spatially and functionally well-separated. To address challenges in translating this approach from rodents to primates, we developed a mouse-to-marmoset pipeline. We first validated that optogenetic targeting of inhibitory neurons (AAV9-Dlx-ChR2) effectively silenced local cortical areas in marmosets. We then tested selective excitation and inhibition of defined projection pathways. By intersecting retrogradely delivered Cre-recombinase (AAVretro-Cre) with locally injected Cre-dependent opsins (AAV8-FLEx-ChR2 or Jaws), we achieved efficient, direction-specific labeling of both callosal and longitudinal projections. This intersectional strategy enabled precise excitatory and inhibitory control of cortical activity using distinct light wavelengths, advancing projection-specific optogenetics for investigating primate brain circuit function.

摘要

灵长类动物的大脑皮层依靠长距离连接来整合功能特化区域之间的信息。研究这些过程需要能够选择性调节特定投射通路的工具。虽然细胞类型特异性光遗传学可以调节局部回路,但这些方法往往缺乏通路特异性。投射特异性光遗传学具有更高的精确性,尤其是在灵长类动物中,其皮层区域在空间和功能上分隔良好。为了解决将这种方法从啮齿动物转化到灵长类动物过程中遇到的挑战,我们开发了一套从小鼠到狨猴的流程。我们首先验证了对抑制性神经元进行光遗传学靶向(AAV9-Dlx-ChR2)能有效沉默狨猴的局部皮层区域。然后我们测试了对特定投射通路的选择性兴奋和抑制。通过将逆行递送的Cre重组酶(AAVretro-Cre)与局部注射的Cre依赖性视蛋白(AAV8-FLEx-ChR2或Jaws)相交,我们实现了对胼胝体投射和纵向投射的高效、方向特异性标记。这种交叉策略能够使用不同的光波长对皮层活动进行精确的兴奋性和抑制性控制,推动了用于研究灵长类动物脑回路功能的投射特异性光遗传学的发展。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/94e8/12360341/489f254dc22c/nihpp-2025.06.18.660378v2-f0002.jpg

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