Developing innovative tools to investigate the anatomy and molecular profile of striatal cholinergic interneurons

摘要 纹状体是大脑皮层下的一个区域，对动作选择、学习和运动行为至关重要。 纹状体胆碱能中间神经元（ChI）是局部乙酰胆碱的主要来源，并在神经元中起作用。 在调节纹状体回路中的重要作用。因此，在许多情况下， 涉及纹状体功能障碍的神经精神障碍，如帕金森病、阿尔茨海默病、 疾病和肌张力障碍。尽管其重要性，知之甚少ChI的结构和功能。Chi是 一种稀疏细胞类型，仅占所有纹状体神经元的~1%，但具有巨大的领土影响力 通过发出密集的投影。ChIs的完整形态和局部网络解剖仍然存在 未知此外，最近的一项研究表明，ChI并不像以前认为的那样是同质的， 根据形态学和分子标记可分为亚型。然而，相关性 ChI解剖学和分子图谱之间的关系还有待阐明。 这项建议旨在扩大我们的理解ChI解剖和分子概况， 开发新的工具来回答以前无法解决的问题。虽然以前 我的初步数据表明， 一种新的ChI形态亚型：棘状ChI。我将描述多刺的完整形态， 将Brainbow与新的清理和扩展技术相结合。然后我将开发 技术，同时可视化形态和查询ChIs的分子谱， 把两者联系起来这些研究将增加我们对ChI细胞类型的理解，并揭示 它们的解剖学和分子概况如何有助于功能的基本原理。新工具 本项目中开发的软件也将传播给神经科学界。最后， 完成这份建议书将有助于推动我实现成为一名医生科学家的长期目标 通过为我提供严格的科学、技术和临床培训。

Abstract The striatum is a subcortical brain region critical for action selection, learning, and motor behavior. Striatal cholinergic interneurons (ChIs) are the primary source of local acetylcholine, and play an important role in modulating striatal circuits. Consequently, ChIs are dysregulated in many neuropsychiatric disorders that involve striatal dysfunction, such as Parkinson's disease, Alzheimer's disease, and dystonia. Despite their importance, little is known about ChI structure and function. ChIs are a sparse cell type, comprising only ~1% of all striatal neurons, yet have tremendous territorial influence by sending out dense projections. The complete morphology and local network anatomy of ChIs remain unknown. Furthermore, a recent study suggested ChIs are not homogenous as previously believed, but may be divided into subtypes based on morphology and molecular markers. However, the correlation between ChI anatomy and molecular profiles has yet to be elucidated. This proposal seeks to expand our understanding of ChI anatomy and molecular profiles by developing new tools to answer questions that were impossible to address before. Although previously understood to be an aspiny, homogenous population, my preliminary data demonstrates the existence of a new ChI morphological subtype: spiny ChIs. I will characterize the complete morphologies of spiny and aspiny ChIs by combining Brainbow with new clearing and expansion technology. I will then develop technology to simultaneously visualize morphology and interrogate the molecular profile of ChIs in order to correlate the two. These studies will increase our understanding of ChI cell types, and reveal fundamental principles of how their anatomy and molecular profiles contribute to function. The novel tools developed in this project will also be disseminated for the benefit of the neuroscience community. Finally, completing this proposal will help drive me towards my long-term goal of becoming a physician scientist by providing me with rigorous scientific, technical, and clinical training.