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

摘要 纹状体是大脑皮质下的一个区域，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 在调节纹状体回路中起着重要作用。因此，在许多情况下，CHI是不受监管的 涉及纹状体功能障碍的神经精神障碍，如帕金森氏症、阿尔茨海默氏症 疾病和肌张力障碍。尽管它们很重要，但人们对CHI的结构和功能知之甚少。池子是 一种稀疏的细胞类型，仅占所有纹状体神经元的1%，但具有巨大的地域影响力 通过发出密集的投射。保留了CHI的完整形态和局部网络解剖。 未知。此外，最近的一项研究表明，CHI并不像之前认为的那样是同质的，但 根据形态和分子标记可分为亚型。然而，这种关联性 CHI的解剖结构和分子图谱之间的关系尚不清楚。 这项建议旨在通过以下方式扩大我们对CHI解剖学和分子图谱的理解 开发新的工具来回答以前无法解决的问题。尽管之前 据了解，这是一个志同道合的群体，我的初步数据证明了 一种新的气形态亚型：多刺智齿。我将描述多刺的完整形态和 通过将Brain弓与新的清算和扩展技术相结合，实现了精明的CHIS。然后我就会发展 有序地同时可视化形态和查询CHI分子轮廓的技术 将两者联系起来。这些研究将增加我们对CHI细胞类型的理解，并揭示 它们的解剖结构和分子特征如何影响其功能的基本原理。新奇的工具 还将为神经科学界的利益而传播在该项目中开发的信息。最后， 完成这项提议将有助于我朝着成为一名内科科学家的长期目标前进 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.