Dissection of Cell Type Specific Contributions to Motor Learning Circuits

项目摘要 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，我们的许多人 日常动作需要熟练准确的动作。然而，为了达到熟练程度，这些运动技能必须 首先是通过运动学习的过程来学习。关于这一主题的许多工作都集中在动力学上 在运动系统的不同部分有不同种类的神经元。然而，具体的类型是否 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在功能上发生变化 彻底调查。回答这些问题是理解马达电路如何进化的关键一步 过度学习以提高运动能力。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 想想运动疾病，特别是那些有运动学习障碍的人，比如自闭症谱系障碍。 在这项建议中，我首先研究第一个领域--运动学习的细胞决定因素。通过专注于 初级运动皮质，运动系统的中央协调器，运动技能所必需的区域 学习，我首先问不同的细胞类型是否比运动学习更丰富，这项研究与 《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， 在前肢任务中标记活跃的细胞，并允许分离它们和单细胞转录图谱。这 使我能够比学习更好地检验细胞类型的特异性丰富。在目标2中，也是在指导阶段 (K99)，我也开始探索第二个领域，通过检查特定细胞类型内的潜在变化，我 在学习后期，FoxP2在M1的第VI层表达细胞。这些细胞投射到马达 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 学习。我绘制了这些细胞的全脑投影模式，检查了它们的动态和相互作用 学习，扰乱他们的活动。最后，在目标3的独立阶段(R00)，我继续讨论这两个问题 关于细胞类型、特定贡献和细胞内在变化的问题，但在全脑水平上。通过使用Fos，我将 确定在学习和经历转录变化时参与的区域。我将探讨几个角色 通过功能操作和通过学习识别活跃的细胞类型来识别区域。那我会的 通过在体内进行钙动力学的单细胞成像，将细胞类型和分子变化结合起来 和Fos的表达，以探讨神经元活动与诱导转录变化的关系。 鉴于我在分子生物学和神经科学方面的培训，我处于一个独特的位置来进行这项工作 两个字段的交集。我很幸运地在哥伦比亚大学的扎克曼研究所，一个合作的 以及身临其境的神经科学中心。与我的导师和共同导师鲁伊·科斯塔博士和伊丽莎白·希尔曼博士一起， 他在系统神经科学和行为学方面的专长与我在分子生物学方面的背景相辅相成，我计划 继续我在系统神经科学方法、转录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.