Establishment of the mothod for RNA labelling in a neuron subtype-specific manner in vivo

2020 Fiscal Year Final Research Report

• Project/Area Number: 19K22580
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 51:Brain sciences and related fields
• Research Institution: Osaka University
• Principal Investigator: Kawahara Yukio 大阪大学, 医学系研究科, 教授 (80542563)
• Co-Investigator(Kenkyū-buntansha): 加藤 有己 大阪大学, 医学系研究科, 助教 (10511280)
• Project Period (FY): 2019-06-28 – 2021-03-31
• Keywords: 遺伝子発現 / RNAラベリング / マウス / 中枢神経

Outline of Final Research Achievements

Selective degeneration of certain specific neurons is a common feature found in neurodegenerative diseases. However, the causative mechanisms remain elusive, which hampers the understanding of disease pathogenesis and the establishment of therapeutic strategies. In this context, it is crucial to characterize individual neuronal subtypes in terms of their RNA expression profiles. In this study, we challenged ourselves to develop a novel RNA labeling method to collect RNAs from certain specific neurons with high accuracy. We chose cerebellar Purkinje cells as a control and examined various labeling conditions and RNA recovery methods. As a result, we were able to confirm the presence of marker genes expressed in Purkinje cells. However, the background noise caused by contaminated non-specific RNAs remains high, which needs further improvement. We will continue to devise sequencing methods and bioinformatics analyses for establishing the corresponding method with higher accuracy.

Free Research Field

神経変性疾患

Academic Significance and Societal Importance of the Research Achievements

近年、単一細胞や単一核の遺伝子発現を網羅的に解析できるようになった。その一方で、分離せずに生体内の特定の細胞だけの遺伝子発現情報を得ることは依然として困難である。また、神経細胞については軸索やシナプスに分布するRNAの情報を得ることはできない。本手法が実現されれば、それぞれの細胞種の遺伝子発現プロファイルを特徴付けることができる。また、疾患モデルマウスに適用すれば、パーキンソン病やALS発症の鍵を握る遺伝子を同定できる可能性もある。その結果として治療法の確立へと繋がることも期待できることから、高い意義をもった内容だと考える。