公募シンポジウム

公募シンポジウム10【最先端技術をつかって神経系細胞の生涯を理解する】

2021/10/1　10:00～12:00　ZOOM B会場 S10-1 神経幹細胞運命決定機構を明らかにする1 細胞遺伝子発現・エピゲノム解析 Single cell RNA-seq and ATAC-seq analysis identified the mechanism underlying neural stem cell fate decisions ^○松田泰斗,松原周蔵,中島欽一九州大学医学研究院基盤幹細胞学分野 ^○Taito Matsuda, Shuzo Matsubara, Kinichi Nakashima Department of Stem Cell Biology and Medicine, Graduate School of Medical Sciences, Kyushu University Neural stem cells (NSCs) in the adult hippocampus generate new neurons via a process referred to as neurogenesis, supporting cognitive functions. Since altered neurogenesis has been reportedly associated with several diseases, understanding the molecular basis of NSC activity is an important focus for developing a therapeutic strategy. During aging, the ability of NSCs to proliferate and give rise to new neurons decreases dramatically. The decrease in neurogenesis with age is accompanied by poorer performance on learning and memory tasks. However, the mechanisms involved in this age-dependent reduction of NSC activity remain elusive. We surveyed the single-cell transcriptomic and chromatin landscape of hippocampal NSCs during the aging process using Nestin-EGFP mice and identified the alterations of NSC-specific gene expression signatures and accessible chromatin state. We further revealed that the age-dependent downregulation of SETD8, a methyltransferase that catalyzes the mono-methylation of histone H4 lysine 20, underlies age-related alterations in adult hippocampal NSC proliferation and neurogenesis. In this talk, we will introduce these recent findings of the mechanism underlying neural stem cell fate decisions during the aging process and discuss the application of single-cell technology to assess behavioral changes of the brain cells, including NSCs.		2021/10/1　10:00～12:00　ZOOM B会場 S10-2 質量分析を基盤とした神経幹細胞のマルチオミクス解析 Multi-omics analysis of neural stem cells based on mass spectrometry ^○和泉自泰九州大学生体防御医学研究所メタボロミクス分野 ^○Yoshihiro Izumi Division of Metabolomics, Medical Institute of Bioregulation, Kyushu University Each omics analysis is used in various research fields, and it is expected that integrated analysis of genome, transcriptome, proteome, and metabolome will lead to a true understanding of cell function. Proteins and metabolites are important molecules in complex biological systems. For example, proteins have many molecules that are activated by post-translational modifications. Similarly, molecules that control the activity of DNA, RNA, and proteins have been found in metabolites. Therefore, at present, it is not possible to predict changes in proteome and metabolome from gene expression information. On the other hand, since proteins and metabolites have various physicochemical properties, mass spectrometry is an essential tool in proteome and metabolome analyses due to its selectivity and sensitivity. We have developed metabolome and proteome analytical techniques based on chromatography mass spectrometry. In this talk, I will introduce "search for factors involved in the dormant state of neural stem cells by multi-omics analysis" and "current status of single-cell metabolome and proteome analyses".

2021/10/1　10:00～12:00　ZOOM B会場 S10-3 光照射した領域に限定した高深度空間トランスクリプトーム法の開発 High-depth spatial transcriptomics technology via photo-isolation chemistry ^○本田瑞季¹,木村龍一¹,原田哲仁²,前原一満²,田中かおり²,大川恭行²,沖真弥¹ 1.京都大学大学院医学研究科創薬医学講座,2.九州大学生体防御医学研究所トランスクリプトミクス分野 ^○Mizuki Honda¹,Ryuichi Kimura¹,Akihito Harada²,Kazumitsu Maehara²,Kaori Tanaka²,Yasuyuki Ohkawa²,Shinya Oki¹ 1. Department of Drug Discovery Medicine, Kyoto University Graduate School of Medicine,2. Division of Transcriptomics, Medical Institute of Bioregulation, Kyushu University To gain insights into tissue-specific gene expression in multicellular systems, spatial information is required and precisely linked with gene expression profiles. Here, we have developed a novel high-depth spatial transcriptomics technology, photo-isolation chemistry (PIC), which is able to isolate gene expression profiles only from photo-irradiated region out of whole tissues. The most important key to this technology is caged oligo DNA modified with photo-cleavable blocker. We performed reverse transcription on tissue section of mouse embryo using caged oligo DNA, and then photo-irradiated on to three narrow regions (φ75 μm spotlight onto ~80 cells in ventral, dorsal, or medial regions). As a result, more than 10,000 genes were detected from photo-irradiates region. Hundreds of genes were detected to be expressed in a manner specific for photo-irradiated regions, some of which were validated by in situ hybridization. Furthermore, to apply PIC for free-form ROIs, we performed PIC RNA-seq by photo-irradiating the CA1 region, CA3 region, and dentate gyrus of the adult mouse hippocampus using a multi-pattern illumination system with a digital mirror device, and successfully detected region-specific genes. Thus, this technology enables us to isolate transcriptomic profiles from region of interest with high spatial resolution and detection sensitivity.		2021/10/1　10:00～12:00　ZOOM B会場 S10-4 神経伝達物質の量的変化を質量分析で可視化する試み Department of Biomedical Sciences, School of Biological and Environmental Sciences, Kwansei Gakuin University ^○矢尾育子関西学院大学生命環境学部生命医科学科 ^○Ikuko Yao Department of Biomedical Sciences, School of Biological and Environmental Sciences, Kwansei Gakuin University Neurotransmitters play crucial roles in the brain functions and regulate various biological processes. Abnormal concentrations of neurotransmitters and consequent dysfunction are linked to various central nervous system disorders. Especially, glutamate and gamma-aminobutyric acid (GABA) have pivotal roles in central nervous system and closely involves severe neurological disease such as epilepsy. Visualization of the concentrations of neurotransmitters is thought to be essential in understanding their role in various neurophysiological processes in different regions of the brain. The detection of low-molecular weight components such as amino neurotransmitters by matrix-assisted laser desorption/ionization-imaging mass spectrometry (MALDI-IMS) has been tried and developed by some groups. For example, several chemical derivatization methods have been made it possible to detect them on tissues in recent years. In this session, we will introduce some examples visualization of neurotransmitters by IMS and discuss the application and future development of this method. We applied this technique to investigate Scrapper-knockout (SCR-KO) mice. SCRAPPER is a synaptic protein which we have identified as an ubiquitin E3 ligase. SCRAPPER is involved in the ubiquitination of RIM (Rab3-interacting molecule) 1, an important regulator of synaptic plasticity, and thus regulates synaptic transmissions. SCR-KO has the defect in neurotransmission via excessive secretion of neurotransmitters due to the upregulation of the release probability. IMS with on tissue derivatization revealed that SCR-KO mice had significant upregulation of glutamate and GABA levels in multiple regions. The alteration visualized would reflect the defect in neurotransmission in the SCR-KO mouse brain.