Dissection of Cell Type Specific Contributions to Motor Learning Circuits

工程摘要 无论是在狭窄的小路上骑自行车，还是在小盒子里拿你最喜欢的饼干， 日常动作需要熟练和准确的动作。然而，要达到熟练程度，这些运动技能必须 首先是通过运动学习的过程来学习。关于这个问题的许多工作都集中在动力学上， 在运动系统的不同部分的异质神经元群体。但是，具体类型 的神经元在学习过程中被招募和/或神经元在学习过程中是否发生功能性变化还没有被证实。 彻底探索。回答这些问题是理解电动机电路如何演变的关键一步 以提高运动能力。因此，我的长期目标是专注于（1）细胞 类型特异性贡献和（2）运动学习过程中细胞类型的内在变化。这将有助于指导我们的 思考运动疾病，特别是那些运动学习缺陷的人，比如自闭症谱系障碍。 在这个建议中，我开始检查第一个领域-运动学习的细胞决定因素。通过关注 初级运动皮层，运动系统的中央协调器和运动技能所必需的区域 学习，我开始问是否不同类型的细胞丰富的运动学习，一项研究符合 《大脑2025》报告的第一个目标。在目标1中，在指导阶段（K99），我提出了一种新的方法， 在前肢任务期间标记活性细胞，并允许它们的分离和单细胞转录谱分析。这 让我能够研究细胞类型特异性富集在学习过程中的作用。在目标2中，也是在辅导阶段 (K99)，我开始也通过检查特定细胞类型内的潜在变化来探索第二个领域， 鉴定为在后期学习时富集的，M1的第VI层中的FoxP 2表达细胞。这些细胞投射到发动机 丘脑，另一个与运动学习密切相关的区域，使它们成为进一步研究的有趣候选者。 study.我绘制了这些细胞的全脑投射模式，检查了它们的动力学和参与度， 学习，扰乱他们的活动。最后，在目标3中，在独立阶段（R 00），我继续解决这两个问题。 细胞类型特异性贡献和细胞内在变化的问题，但在全脑水平。通过使用Fos，我将 识别在学习过程中参与并经历转录变化的区域。我将探讨几个角色 通过功能操作和识别活动细胞类型来识别区域。然后我将 通过进行钙动力学和细胞形态学的体内单细胞成像来整合细胞类型和分子变化 和Fos表达，探讨神经元活性与诱导的转录变化之间的关系。 鉴于我在分子生物学和神经科学方面的训练，我处于一个独特的位置，可以在 两个领域的交集。我很幸运能在哥伦比亚大学的朱克曼研究所工作， 和神经科学的中心沿着我的导师和共同导师，鲁伊·科斯塔博士和伊丽莎白·希尔曼博士， 他在系统神经科学和行为学方面的专业知识补充了我在分子生物学方面的背景，我计划 继续我在系统神经科学方法，转录组数据分析和实验室管理方面的培训。

Project Abstract Whether riding your bike down a narrow path or reaching for your favorite cookie in a small box, many of our daily actions require skilled and accurate movements. However, to achieve proficiency, these motor skills must first be learned through the process of motor learning. Much work on this subject has focused on the dynamics of heterogeneous populations of neurons in various parts of the motor system. However, whether specific types of neurons are recruited over learning and/or whether neurons change functionally over learning has not been thoroughly explored. Answering these questions is a key step in understanding how the motor circuit evolves over learning to allow for increased motor proficiency. Therefore, my long-term objective is to focus on (1) cell type specific contributions and (2) cell type intrinsic changes during motor learning. This will help instruct our thinking about motor diseases, especially those with motor learning deficits like autism spectrum disorder. In this proposal, I begin by examining the first area - the cellular determinants of motor learning. By focusing on the primary motor cortex, a central coordinator of the motor system and a region necessary for motor skill learning, I begin by asking whether different cell types are enriched over motor learning, a study in line with the first aim of the BRAIN 2025 report. In aim 1, during the mentored phase (K99), I present a novel approach that tags active cells during a forelimb task and allows for their isolation and single-cell transcriptional profiling. This has allowed me to examine cell type specific enrichment over learning. In aim 2, also during the mentored phase (K99), I begin to also explore the second area by examining potential changes within a specific cell type which I identified as enriched at late learning, FoxP2 expressing cells in layer VI of M1. These cells project to the motor thalamus, another region strongly implicated in motor learning, making them an interesting candidate for further study. I map the brain-wide projection pattern of these cells, examine their dynamics and engagement over learning, and perturb their activity. Finally, in aim 3, in the independent phase (R00), I continue to address both questions of cell type specific contribution and cell intrinsic changes but at a brain-wide level. By using Fos, I will identify regions engaged during learning and undergoing transcriptional changes. I will explore the role of several identified regions with functional manipulations and by identifying the active cell types over learning. I will then integrate both cell type and molecular changes by conducting in vivo single cell imaging of both calcium dynamics and Fos expression to explore the relationship between neuronal activity and induced transcriptional changes. Given my training in molecular biology and neuroscience, I am in a unique position to conduct this work at the intersection of both fields. I am fortunate to be at the Zuckerman Institute at Columbia University, a collaborative and immersive hub of neuroscience. Along with my mentor and co-mentor, Drs. Rui Costa and Elizabeth Hillman, whose expertise in systems neuroscience and behavior complement my background in molecular biology, I plan to continue my training in systems neuroscience approaches, transcriptomic data analysis, and lab management.