Local Connections and in Vivo Physiology of Inhibitory Cortical Neurons.

描述（由申请人提供）：这是我的NIH独立之路奖（4 R 00 DA 023700）倾斜“抑制性皮质神经元的局部连接和体内生理学”的竞争性修订申请，以响应NIH资助通知（NOT-OD-09-058，NIH宣布恢复法案资金的可用性竞争性修订申请）。尽管对皮层回路的基本蓝图有广泛的了解，但对局部皮层回路、特定细胞类型的连接性及其功能的详细了解仍然有限。最初提出的研究调查了兴奋性和抑制性突触输入到大脑皮层中特定类型的抑制性神经元的层状和精细尺度特异性，并检查了不同抑制性细胞类型的体内生理学及其对皮层活动的参与和调节。我们已经完成的目的，检查层特异性的功能输入不同的细胞类型相结合的全细胞记录与扫描激光光刺激在脑切片。我们在了解特定抑制性细胞类型的体内生理学方面也取得了进展。最近，我们已经开发出一种技术，通过电压敏感染料成像和激光扫描光刺激的新组合，使高分辨率和快速功能成像的脑切片。这一创新将在皮层电路研究领域产生广泛的影响，因为它有助于快速映射和精确评估皮层组织和功能。该创新将能够通过检查抑制性细胞类型对光刺激诱发的神经元群体活动的影响并通过电压敏感染料成像检测来映射抑制性细胞类型的抑制输出。鉴于我最初的项目取得了令人兴奋的进展，我想修改我最初的目标，专注于进一步改进新技术，并将这种技术扩展到阐明皮层回路。新修订的具体目标是（1）完善和改进脑切片中的高分辨率和快速功能成像技术;（2）使用新技术绘制特定类型抑制性神经元的抑制性输出;（3）使用新技术表征多巴胺受体敲除（D2 R-/-）小鼠中的回路改变。这一修订要求支持购买更多设备和培训一名少数民族研究生。 公共卫生相关性：研究涉及特定抑制细胞类型的皮层回路的详细组织对于理解皮层功能是必要的。这些研究对人类健康具有重要意义，因为这些细胞类型及其活动涉及许多疾病模型，并且可以介导与药物成瘾和滥用相关的皮质机制。

DESCRIPTION (provided by applicant): This is a competitive revision application for my NIH Pathway to Independence Award (4 R00DA023700) tilted "Local Connections and In Vivo Physiology of Inhibitory Cortical Neurons", in response to the NIH funding notice (NOT-OD-09-058, NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications). Despite extensive knowledge of the basic blueprint of cortical circuits, detailed knowledge about local cortical circuits, the connectivity of specific cell types and how they function is still limited. The studies originally proposed investigate the laminar and fine-scale specificities of excitatory and inhibitory synaptic input to specific types of inhibitory neurons in the cerebral cortex, and examine in vivo physiology of distinct inhibitory cell types and their participation and regulation of cortical activities. We have accomplished the aims of examining the laminar specificity of functional input to distinct cell types by combining whole cell recordings with scanning laser photostimulation in brain slices. We also have progressed toward understanding in vivo physiology of specific inhibitory cell types. Recently, we have developed a technique enabling high-resolution and fast functional imaging in brain slices through a novel combination of voltage sensitive dye imaging and laser scanning photostimulation. This innovation will have broad impacts in the field of cortical circuitry studies, as it facilitates rapid mapping and precise evaluation of cortical organization and function. The innovation will enable the ability to map inhibition output from inhibitory cell types by examining their influence on neuronal population activities evoked by photostimulation and detected by voltage sensitive dye imaging. Given the exciting developments of my original projects, I would like to revise my original aims, and focus on further improving the new technique and extending such technology toward elucidating cortical circuitry. The newly revised Specific Aims are to (1) Refine and improve the high-resolution and fast functional imaging technique in brain slices; (2) Map inhibitory output of specific types of inhibitory neurons using the novel technique; (3) Characterize circuit alterations in dopamine receptor knockout (D2R-/-) mice with the novel technique. This revision requests support to purchase additional equipment and train one minority graduate student. PUBLIC HEALTH RELEVANCE: Studies of the detailed organization of cortical circuits involving specific inhibitory cell types are necessary toward understanding cortical function. These studies have important implications for human health, as these cell types and their activities are involved in many disease models and can mediate the cortical mechanisms related to drug addiction and abuse.