公募シンポジウム

公募シンポジウム多様な神経疾患に対する組織培養研究の新展開

7月6日（木）　16:30-18:30　Room F 1SY⑩-1 培養細胞から学ぶ生命原理と神経変性原理 Life and neurodegeneration fundamentals revealed by cell culture 岡澤　均東京医科歯科大学　神経病理学 Hitoshi Okazawa Dept. of Neuropathol., Tokyo Medical and Dental University, Tokyo, Japan Cell culture is an essential technique for researches in basic and clinical sciences. We have been using cell lines, primary neurons, primary microglia and iPS cell-derived neurons for our researches that is aimed for understanding and therapeutic development of neurodegenerative diseases. In this talk, I would like to introduce you four typical cases in our research topics (if time allows). The first one is discovery of an essential factor for nucleolus, the key cell organella for ribosomal RNA production and for all cell functions in life. In this case, we used cell line to show fine structure of nucleolus by super resolution microscopy, electron microscopy and CLEM. The second one is discovery of a new mechanism for recognition of Tau, a neurodegenerative protein by microglia culture, in which we used primary culture of microglia. The third case is discovery of Oct-3/4, the transcription factor at the top of the molecular cascade for development. In this case that lead to Nobel prize of Prof Yamanaka, we used pluripotent stem cells. The final case is discovery of a new prototype of neurodegenerative necrosis, in which we used iPSC-derived neurons. I hope our experiences would become some hints for young neuroscientists and neuropathologists.		7月6日（木）　16:30-18:30　Room F 1SY⑩-2 ヒト脳発生と神経発達障害を研究するためのヒト組織培養法 Human tissue culture methods for studying human brain development and neurodevelopmental disorders 難波　隆志ヘルシンキ大学　HiLIFE Takashi Namba Neuroscience Center, HiLIFE, University of Helsinki Human has a neocortex that has characteristically expanded and convoluted. Several approaches have been utilized to examine the developmental process of human neocortex.In this symposium, we describe three neocortical tissue culture methods that can be combined with genetic manipulation by electroporation and treatment with specific inhibitors:(i) organotypic slice culture; (ii) free-floating tissue culture hemisphere rotation culture; and (iii) cerebral organoid culture. Each of these three culture methods offers distinct features with regard to the analyses to be performed. These three culture methods are therefore powerful techniques to examine the function of genes involved in human neocortical development.

7月6日（木）　16:30-18:30　Room F 1SY⑩-3 Differentiation dynamics of oligodendrocyte progenitor cells in culture and in vivo 竹林　浩秀新潟大学　神経解剖 Hirohide Takebayashi Division of Neurobiol and Anatomy, Niigata Univ, Niigata, Japan Oligodendrocytes are glial cells of the central nervous system that form myelin. Oligodendrocytes are formed through terminal differentiation from oligodendrocyte progenitor cells (OPCs) to mature oligodendrocytes. Cell culture experiments have revealed many aspects of oligodendrocyte differentiation mechanisms using primary cultures of OPCs. In addition, lineage tracing experiments using Olig2-CreER mice were performed to investigate in vivo differentiation of OPCs in various conditions. We will discuss the usefulness and points to consider for both culture and cell lineage tracing experiments.		7月6日（木）　16:30-18:30　Room F 1SY⑩-4 新規ヒト血液脳関門in vitroモデル Novel human in vitro blood-brain barrier model 神田　隆山口大学　神経・筋難病治療学 Takashi Kanda Dept. of Neurotherapeutics, Yamaguchi Univ. School of Medicine, Ube, Japan Destruction of blood-brain barrier (BBB) is the initial key step in the development of neuroimmunological disorders of the central nervous system such as multiple sclerosis and meuromyelitis optica. Several laboratory developed cell lines of BBB composing cells and in vitro models; however, to address this pertinent issue, we propose some important properties that will be necessary for an in vitro BBB model that is more robust than those currently available. To construct in vitro BBB models which retain in vivo properties, we used human ECs (TY10), human astrocytes with AQP4 expression, and human pericytes, each of which was conditionally immortalized by transfection with temperature-sensitive SV40 (simian virus 40) large T antigen (ts-SV40-LT) and human telomerase gene which retains both their physiologic and morphologic BBB properties. Further, we constructed functional in vitro static and ex vivo flow-based models using the newly established triple coculture system. The new static in vitro model allowed long-term measurement of transepithelial electrical resistance (TEER) and measurement of microvolumes of IgG translocation through the BBB. The new ex vivo flow-based model enabled us to evaluate leukocyte transmigration across the BBB. In this symposium, recent achievements using this system will be presented.

7月6日（木）　16:30-18:30　Room F 1SY⑩-5 神経再生促進と神経変性抑制における神経回路形成因子LOTUSのシナプス形成制御 Regulation of synapse formation by LOTUS in promoting nerve regeneration and suppressing neurodegeneration 竹居　光太郎^1,2 1. 横浜市立大学　医学部　臓器再生医学教室　神経再生医学講座, 2. 横浜市立大学大学院　生命医科学研究科 Kohtaro Takei^1,2 1. Lab. for Neural Reg. Med., Dept. of Reg. Med., Yokohama City Univ. Sch. of Med., Yokohama, Japan, 2. Dept. of Med. Life Sci., Yokohama City Univ. Grad. Sch. of Med. Life Sci., Yokohama, Japan Nogo and amyloid beta (Aβ) proteins have been reported to reduce memory via a decreased synaptic density. The lateral olfactory tract usher substance (LOTUS) is an endogenous antagonist of Nogo receptor-1(NgR1) and paired immunoglobulin-like receptor B (PirB). We found that loss of LOTUS impairs memory via decreased synaptic density in the hippocampus. We examined if LOTUS overexpression induced synapse formation and memory. An increased synaptic density was found in the hippocampus of LOTUS-overexpressing transgenic (LOTUS-Tg) mice, and the LOTUS-Tg mice showed enhanced hippocampus-dependent memory. Furthermore, we found that LOTUS inhibited the binding of Aβ to PirB and the activation of downstream signaling of PirB, thereby giving that LOTUS inhibited Aβ-induced decrease in spine density in cultured hippocampal neurons. These findings suggest that LOTUS may promote synapse formation by antagonistic action to Nogo protein and suppress synapse elimination by blockade of Aβ-induced neurodegeneration.		7月6日（木）　16:30-18:30　Room F 1SY⑩-6 人為的シナプスコネクトによる慢性期脊髄損傷回復への試み Recovery from the chronic stage spinal cord injury by synthetic synaptic organizer. 武内　恒成^1,2, 笹倉　寛之¹, 池野　正史¹, 柚崎　通介³ 1. 愛知医科大学医学部細胞生物学, 2. 愛知医科大学　研究創出センター, 3. 慶応義塾大学医学部生理学 Kosei Takeuchi^1,2, Hiroyuki Sasakura¹, Masashi Ikeno¹, Michisuke Yuzaki³ 1. Dept.Cell Biol.,Aichi Medical Univ. Japan, 2. Medical Research Creation Center, Aichi Medical University, 3. Dept. fo Physiology, Keio Univ Sch.of Med. Spinal cord injury(SCI) causes permanent dysfunction in the body movement, and the effective treatment to cure the chronic stage is not well known. We previously showed that the synthesized synapse organizer, CPTX, restored the locomotion of acute phase SCI in mice (Science, 2020). CPTX is a synthetic chimeric protein, which connects the pre-synaptic neurexin and the post-synaptic AMPA receptors in vitro and in vivo. We next asked if the CPTX is effective to chronic stage. The single injection of CPTX on chronic phase activated the hind leg movement, which otherwise showed the permanently poor movement. To achieve more robust recovery, we are combining with rehabilitation. We are also developing the AI-based behavioral evaluation systems and the single nuclear RNA-seq gene expression profiling to gain insights into CPTX induced recovery.