成体脳のニューロン再生における死細胞の貪食過程とその意義

• Project Area: Homeostatic Regulation by Various Types of Cell Death
• Project/Area Number: 17H05512
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Nagoya City University
• Principal Investigator: 澤本 和延 名古屋市立大学, 大学院医学研究科, 教授 (90282350)
• Project Period (FY): 2017-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥9,620,000 (Direct Cost: ¥7,400,000、Indirect Cost: ¥2,220,000); Fiscal Year 2018: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2017: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
• Keywords: 再生医学 / 神経科学 / 脳・神経

Outline of Annual Research Achievements

脳に存在するニューロン（神経細胞）の大部分は、胎生期あるいは生後の発達期に作られる。しかし、多くの動物種において、脳内で外界からの嗅覚情報を処理する嗅球においては、成体においても継続的にニューロンの細胞死とニューロン再生が起こっていることが知られている。以前研究代表者らは、二光子顕微鏡を用いて、嗅球ニューロンが細胞死によって除去され、同じ場所に同じ種類のニューロンが再生するという嗅覚入力依存的なニューロン再生メカニズムを見出した。
本課題では、二光子イメージング等を用いて、ミクログリアが死んだニューロンを除去し、新しいニューロンの再生を助ける生理的なメカニズムの詳細を明らかにすることを目的としている。平成30年度は以下の実験を実施した。
1) ミクログリアの貪食動態と機能：ミクログリアは死細胞を貪食することが知られているが、生後脳における新生ニューロンの貪食動態とその意義については不明な点が多い。昨年度に引き続き、嗅球内の死細胞とその貪食過程を、電子顕微鏡等で解析し、細胞死の特徴などを明らかにした。前年度検討した貪食抑制効果を有する分子を応用した実験方法を確立した。
2)ミクログリアが誘導するニューロン再生機構の解明：ミクログリアが単に死んだニューロンを除去するのみならず、積極的にニューロン再生を促進しているという仮説を検討するため、昨年度に引き続き、ミクログリアがニューロン再生を促進する可能性のある分子機構についての解析を行った。

Research Progress Status

平成30年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

平成30年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] バレンシア大学(スペイン)
• [Int'l Joint Research] バレンシア大学/Achucarro Basque Center for Neuroscience(スペイン)
• [Journal Article] Ventricular-subventricular zone fractones are speckled basement membranes that function as a neural stem cell niche. (2019)
  • Author(s): Sato Y, Kiyozumi D, Futaki S, Nakano I, Shimono C, Kaneko N, Ikawa M, Okabe M, Sawamoto K, Sekiguchi K.
  • Journal Title: Molecular Biology of the Cell Volume: 30 Issue: 1 Pages: 56-68
  • DOI: 10.1091/mbc.e18-05-0286
  • NAID: 120006626421
  • Peer Reviewed / Open Access
• [Journal Article] New neurons use Slit-Robo signaling to migrate through the glial meshwork and approach a lesion for functional regeneration. (2018)
  • Author(s): Kaneko N, Herranz-Perez V, Otsuka T, Sano H, Ohno N, Omata T, Nguyen HB, Thai TQ, Nambu A, Kawaguchi Y, Garcia-Verdugo JM, Sawamoto K.
  • Journal Title: Science Advances Volume: 4 Issue: 12 Pages: 0618-0618
  • DOI: 10.1126/sciadv.aav0618
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Heterogeneous distribution of doublecortin-expressing cells surrounding the rostral migratory stream in the juvenile mouse (2018)
  • Author(s): Aoyagi Yuka、Hibi Terumasa、Kimori Yoshitaka、Sawada Masato、Kawakami Ryosuke、Sawamoto Kazunobu、Nemoto Tomomi
  • Journal Title: Journal of Comparative Neurology Volume: 526 Issue: 16 Pages: 2631-2646
  • DOI: 10.1002/cne.24521
  • NAID: 120006734165
  • Peer Reviewed
• [Journal Article] Shootin1b Mediates a Mechanical Clutch to Produce Force for Neuronal Migration. (2018)
  • Author(s): Minegishi T, Uesugi Y, Kaneko N, Yoshida W, Sawamoto K, Inagaki N.
  • Journal Title: Cell Reports Volume: 25 Issue: 3 Pages: 624-639
  • DOI: 10.1016/j.celrep.2018.09.068
  • NAID: 120006537407
  • Peer Reviewed / Open Access
• [Journal Article] Adult neurogenesis and its role in brain injury and psychiatric diseases (2018)
  • Author(s): Hayashi Yoshitaka、Jinnou Hideo、Sawamoto Kazunobu、Hitoshi Seiji
  • Journal Title: Journal of Neurochemistry Volume: 147 Issue: 5 Pages: 584-594
  • DOI: 10.1111/jnc.14557
  • Peer Reviewed / Open Access
• [Journal Article] Go with the Flow: Cerebrospinal Fluid Flow Regulates Neural Stem Cell Proliferation. (2018)
  • Author(s): Kaneko N, Sawamoto K.
  • Journal Title: Cell Stem Cell Volume: 22 Issue: 6 Pages: 783-784
  • DOI: 10.1016/j.stem.2018.05.015
• [Journal Article] Detachment of Chain-Forming Neuroblasts by Fyn-Mediated Control of cell-cell Adhesion in the Postnatal Brain. (2018)
  • Author(s): Fujikake K, Sawada M, Hikita T, Seto Y, Kaneko N, Herranz-Perez V, Dohi N, Homma N, Osaga S, Yanagawa Y, Akaike T, Garcia-Verdugo JM, Hattori M, Sobue K, Sawamoto K.
  • Journal Title: Journal of Neuroscience Volume: 38 Issue: 19 Pages: 4598-4609
  • DOI: 10.1523/jneurosci.1960-17.2018
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] PlexinD1 signaling controls morphological changes and migration termination in newborn neurons (2018)
  • Author(s): Sawada Masato、Ohno Nobuhiko、Kawaguchi Mitsuyasu、Huang Shih‐hui、Hikita Takao、Sakurai Youmei、Bang Nguyen Huy、Quynh Thai Truc、Ishido Yuri、Yoshida Yutaka、Nakagawa Hidehiko、Uemura Akiyoshi、Sawamoto Kazunobu
  • Journal Title: The EMBO Journal Volume: 37 Issue: 4
  • DOI: 10.15252/embj.201797404
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Radial glial fibers support neuronal migration and regeneration after neonatal brain injury. (2018)
  • Author(s): Jinnou H, Sawada M, Kawase K, Kaneko N, Herranz-Perez V, Miyamoto T, Kawaue T, Miyata T, Tabata Y, Akaike T, Garcia-Verdugo JM, Ajioka I, Saitoh S, Sawamoto K.
  • Journal Title: Cell Stem Cell Volume: 22 Issue: 1 Pages: 128-137
  • DOI: 10.1016/j.stem.2017.11.005
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Daple Coordinates Planar Polarized Microtubule Dynamics in Ependymal Cells and Contributes to Hydrocephalus (2017)
  • Author(s): Maki Takagishi, Masato Sawada, Shinya Ohata, Naoya Asai, Atsushi Enomoto, Kunihiko Takahashi, Liang Weng, Kaori Ushida, Hosne Ara, Shigeyuki Matsui, Kozo Kaibuchi,3Kazunobu Sawamoto, and Masahide Takahashi.
  • Journal Title: Cell Reports Volume: 20 Issue: 4 Pages: 960-972
  • DOI: 10.1016/j.celrep.2017.06.089
  • Peer Reviewed / Open Access
• [Journal Article] Unique Organization of the Nuclear Envelope in the Post-natal Quiescent Neural Stem Cells. (2017)
  • Author(s): Arantxa Cebrian-Silla, Clara Alfaro-Cervello, Vicente Herranz-Perez, Naoko Kaneko, Dae Hwi Park, Kazunobu Sawamoto, Arturo Alvarez-Buylla, Daniel A.Lim, Jose Manuel Garcia-Verdugo.
  • Journal Title: Stem Cell Report Volume: 9 Issue: 1 Pages: 203-216
  • DOI: 10.1016/j.stemcr.2017.05.024
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Angiogenic Biomaterials for Brain Ischemia Using Affinity-Immobilized VEGF on Laminin Porous Sponge. (2017)
  • Author(s): Oshikawa M, Okada K, Kaneko N, Sawamoto K, Ajioka I.
  • Journal Title: Advanced Healthcare Materials Volume: 6 Issue: 11
  • DOI: 10.1002/adhm.201700183
  • Peer Reviewed
• [Journal Article] Mechanisms of neuronal migration in the adult brain (2017)
  • Author(s): Kaneko N, Sawada M, Sawamoto K
  • Journal Title: J Neurochem Volume: - Issue: 6 Pages: 835-847
  • DOI: 10.1111/jnc.14002
  • NAID: 10018687194
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] 生後脳における新生ニューロンの移動・分化機構とその操作技術 (2017)
  • Author(s): 澤本和延
  • Journal Title: 科研費NEWS Volume: 3
• [Presentation] Enhanced neuronal migration through activated glia promotes post- stroke neuronal regeneration (2019)
  • Author(s): Kaneko N, Herranz-Perez V, Otsuka T, Sano H, Ohno N, Omata T, Nguyen HB, Thai TQ, Garcia-Verdugo JM, Sawamoto K
  • Organizer: 9th FAOPS 2019
  • Int'l Joint Research
• [Presentation] 脳梗塞部位への新生ニューロンの移動を支持する足場としての血管の形態解析 (2019)
  • Author(s): 岩田英敏、金子奈穂子、大竹杏佳、戸室幸太郎、柴田帝式、村上英樹、澤本和延
  • Organizer: 第18回日本再生医療学会総会
• [Presentation] 新生ニューロンの移動・分布の制御による脳梗塞後の神経再生の促進 (2019)
  • Author(s): 金子奈穂子、澤本和延
  • Organizer: 第18回日本再生医療学会総会
• [Presentation] Neuronal migration: strategies for development, maintenance and repair of the postnatal brain (2019)
  • Author(s): 澤本和延
  • Organizer: Japan-UK Neuroscience Symposium 2019
  • Int'l Joint Research / Invited
• [Presentation] 成体脳内を移動する新生ニューロンの三次元微細構造 (2019)
  • Author(s): 松本真実、澤田雅人、金子奈穂子、Nguyen HB、Thai TQ、大野伸彦、澤本和延
  • Organizer: 第15回成体脳ニューロン新生懇談会
• [Presentation] 脳梗塞部へのニューロンの移動と血管・アストロサイト (2019)
  • Author(s): 金子奈穂子、澤本和延
  • Organizer: The 3rd Neuro-Vascular meeting
• [Presentation] Morphological changes and migration termination in newborn neurons controlled by Sema3E-PlexinD1 signaling (2018)
  • Author(s): Sawada M, Sawamoto K
  • Organizer: The 3rd NCU&HALLYM International Joint Symposium 2018
  • Int'l Joint Research / Invited
• [Presentation] 生後脳における新生ニューロンの移動機構と意義 (2018)
  • Author(s): 澤本和延
  • Organizer: 第41回日本分子生物学会年会
  • Invited
• [Presentation] 成体脳における新生ニューロンの傷害部への移動制御機構とニューロン再生 (2018)
  • Author(s): 金子奈穂子、澤本和延
  • Organizer: 第41回日本分子生物学会年会
  • Invited
• [Presentation] 神経再生過程におけるニューロンの移動 (2018)
  • Author(s): 澤本和延
  • Organizer: 第40回神経組織培養研究会
  • Invited
• [Presentation] Morphological changes and migration termination of newborn neurons controlled by PlexinD1 signaling (2018)
  • Author(s): Sawada M, Ohno N, Kawaguchi M, Huang SH, Hikita T, Sakurai Y, Nguyen HB, Thai TQ, Ishido Y, Yoshida Y, Nakagawa H, Uemura A, Sawamoto K
  • Organizer: Neuroscience 2018
  • Int'l Joint Research
• [Presentation] 生後脳組織の維持・再生における新生ニューロンの移動機構と意義 (2018)
  • Author(s): 澤本和延
  • Organizer: 第91回日本生化学会大会
  • Invited
• [Presentation] Ezrin controls morphology and proliferation of neural stem cells in the postnatal ventricular-subventricular zone (2018)
  • Author(s): Kaneko N, Ishizaki T, Tamura A, Higuchi K, Tsukita S, Sawamoto K
  • Organizer: 第61回日本神経化学会大会
• [Presentation] Reelin-Dab1 signaling controls detachment of chain-forming new neurons by regulating cell-cell adhesion (2018)
  • Author(s): Sawada M, Fujikake K, Kaneko N, Herranz-Perez V, Homma N, Yanagawa Y, Garcia-Verdugo JM, Hattori M, Sawamoto K
  • Organizer: 第61回日本神経化学会大会
• [Presentation] The role of newly-generated astrocytes and blood vessels in scaffold formation for neuroblasts migrating toward the injured region after ischemic stroke (2018)
  • Author(s): Otake K, Iwata H, Shibata T, Kaneko N, Sawamoto K
  • Organizer: 第61回日本神経化学会大会
• [Presentation] 脳の発達・再生過程における新生ニューロンの移動の足場 (2018)
  • Author(s): 澤本和延
  • Organizer: 第61回日本神経化学会大会
  • Invited
• [Presentation] Endogenous and artificial scaffolds for neuronal migration and regeneration in the injured brain (2018)
  • Author(s): 澤本和延
  • Organizer: 5thTERMIS-WC
  • Int'l Joint Research / Invited
• [Presentation] Mechanisms for the proper migration and positioning of newborn neurons in postnatal brain circuits (2018)
  • Author(s): Sawada M, Sawamoto K
  • Organizer: 21thKSBNS Annual Meeting
  • Int'l Joint Research / Invited
• [Presentation] A transient postnatal peak of neurogenesis supports neuronal migration to the non-human primate neocortex (2018)
  • Author(s): Akter M, Kaneko N, Sawamoto K
  • Organizer: 第41回日本神経科学大会
• [Presentation] PlexinD1シグナルは新生ニューロンの形態変化と移動停止を調節する (2018)
  • Author(s): Sawada M, Ohno N, Kawaguchi M, Huang SH, Hikita T, Sakurai Y, Nguyen HB, Thai TQ, Ishido Y, Yoshida Y, Nakagawa H, Uemura A, Sawamoto K
  • Organizer: 第41回日本神経科学大会
• [Presentation] Scaffolds for brain development, maintenance and repair (2018)
  • Author(s): Sawamoto K
  • Organizer: The 5th Eastern Asia Dermatology
  • Int'l Joint Research / Invited
• [Presentation] 放射状グリアを足場にした新生児期脳傷害後のニューロン再生 (2018)
  • Author(s): 神農英雄、澤田雅人、川瀬恒哉、金子奈穂子、Herranz-Perez V、宮本拓哉、川上巧、宮田卓樹、田畑泰彦、赤池敏弘、Garcia-Verdugo JM、味岡逸樹、齋藤伸治、澤本和延.
  • Organizer: 第162回名古屋市立大学医学会例会
• [Presentation] PlexinD1 signaling controls morphological changes and migration termination in newborn neurons (2018)
  • Author(s): Sawada M, Ohno N, Kawaguchi M, Huang SH, Hikita T, Sakurai Y, Nguyen HB, Thai TQ, Ishido Y, Yoshida Y, Nakagawa H, Uemura A, Sawamoto K
  • Organizer: 日韓神経発生学共同シンポジウム2018
  • Int'l Joint Research / Invited
• [Presentation] Aberration of the Soluble protein Tsukushi leads alteration of adult neurogenesis resulting lateral ventricule expansion with neuronal disease (2018)
  • Author(s): Ito N, Riyadh MA, Ito A, Ishtiyaq Ahmad SA, Anam MB, Kanemura Y, Shinmyo Y, Abdulhaleem M. FA, Hatakeyama J, Kiyonari H, Shimamura K, Takahashi Y, Sawamoto K, Ohta K
  • Organizer: 第70回細胞生物学会・第51回日本発生生物学会 合同大会
• [Presentation] 脳の発達・再生過程における新生ニューロンの移動の足場 (2018)
  • Author(s): 澤本和延
  • Organizer: 第123 回日本解剖学会総会・全国学術集会
  • Int'l Joint Research / Invited
• [Presentation] 脳質下帯由来オリゴデンドロサイト前駆細胞の傷害部への移動・分化過程の解析 (2018)
  • Author(s): 山本誠也, 金子奈穂子, 橋本真耶佳, 宮本拓哉, 竹林浩秀, 池中一裕, 澤本和延
  • Organizer: 第17回日本再生医療学会総会
  • Int'l Joint Research
• [Presentation] PlexinD1は新生ニューロンの形態変化と移動停止を調節する (2018)
  • Author(s): Sawada M, Ohno N, Kawaguchi M, Huang SH, Hikita T, Sakurai Y, Nguyen HB, Thai TQ, Ishido Y, Yoshida Y, Nakagawa H, Uemura A, Sawamoto K
  • Organizer: 第11回神経発生討論会
• [Presentation] Scaffolds for neuronal migration and brain repair. (2018)
  • Author(s): Sawamoto K
  • Organizer: Joint-Symposium between HYU and NCU
  • Int'l Joint Research / Invited
• [Presentation] Cellular adhesion between chain-forming new neurons in the normal and injured adult brain. (2017)
  • Author(s): Matsumoto M, Sawada M, Kaneko N, Nguyen HB, Thai TQ, Kumamoto N, Ugawa S, Ohno N, Sawamoto K
  • Organizer: 第１３回成体脳のニューロン新生懇談会
• [Presentation] Netrin-5 is upregulated in the peri-infarct region after middle cerebral artery occlusion. (2017)
  • Author(s): Yamagishi S, Takarada M, Sawada M, Sawamoto K, Hori O, Sato K
  • Organizer: Neuroscience 2017
  • Int'l Joint Research
• [Presentation] 脳に内在する再生機構 (2017)
  • Author(s): 澤本和延
  • Organizer: 第３回川島カンファレンス
  • Invited
• [Presentation] Migration and differentiation of V-SVZ-derived oligodendrocyte progenitors in the injured cortex. (2017)
  • Author(s): Hashimoto M, Kaneko N, Yamamoto S, Miyamoto T, Takebayashi H, Ikenaka K, Sawamoto K
  • Organizer: 第60回日本神経化学大会
  • Int'l Joint Research
• [Presentation] Fyn promotes detachment of chain-forming new neurons by controlling N-cadherin-mediated adherens junction in the postnatal brain. (2017)
  • Author(s): Seto Y, Fujikake K, Sawada M, Hikita T, Kaneko N, Sobue K, Sawamoto K
  • Organizer: 第60回日本神経化学大会
  • Int'l Joint Research
• [Presentation] Three dimensional ultrastructure of new neurons migrating in the normal and injured adult brain. (2017)
  • Author(s): Matsumoto M, Sawada M, Kaneko N, Nguyen HB, Thai TQ, Kumamoto N, Ugawa S, Ohno N, Sawamoto K
  • Organizer: 第60回日本神経化学大会
  • Int'l Joint Research
• [Presentation] Microglial phagocytosis of dying neurons promotes new neuron addition in the adult olfactory bulb. (2017)
  • Author(s): Sawada M, Inada H, Kohsaka S, Nabekura J, Sawamoto K
  • Organizer: 第60回日本神経化学大会
  • Int'l Joint Research
• [Presentation] Netrin-5 is upregulated in the peri-infarct region after middle cerebral artery occlusion. (2017)
  • Author(s): Yamagishi S, Takarada M, Sawada M, Sawamoto K, Hori O, Sato K
  • Organizer: 第60回日本神経化学大会
  • Int'l Joint Research
• [Presentation] 脳に内在する再生メカニズムとその操作技術 (2017)
  • Author(s): 澤本和延
  • Organizer: 第４回包括的緩和医療科学学術研究会/第５回Tokyo疼痛緩和次世代研究会 合同研究会
  • Invited
• [Presentation] Neuronal migration for maintenance and repair of adult brain. (2017)
  • Author(s): Sawamoto K
  • Organizer: 2017ISN-ESN MEETING
  • Int'l Joint Research / Invited
• [Presentation] Blood vessels work as a scaffold for neuronal migration in the post-stroke non-human primate. (2017)
  • Author(s): Akter M, Kaneko N, Hara K, Inoue K, Adachi K, Okano H, Sawamoto K
  • Organizer: 第40回日本神経科学大会
  • Int'l Joint Research
• [Presentation] Relationship between neuronal turnover and blood flow in the adult olfactory bulb. (2017)
  • Author(s): Ogino T, Sawada M, Inada H, Nabekura J, Sawamoto K
  • Organizer: 第40回日本神経科学大会
  • Int'l Joint Research
• [Presentation] Appropriate positioning of new neurons is critical for the functional rewiring of neuronal circuit in the post-stroke brain. (2017)
  • Author(s): Kaneko N, Sawamoto K
  • Organizer: 第40回日本神経科学大会
  • Int'l Joint Research
• [Presentation] Resting microglia eliminate dying neurons by phagocytosis in the adult olfactory bulb. (2017)
  • Author(s): Sawada M, Inada H, Kohsaka S, Nabekura J, Sawamoto K
  • Organizer: 第40回日本神経科学大会
  • Int'l Joint Research
• [Presentation] Migration of new neurons for maintenance and repair of adult brain. (2017)
  • Author(s): 澤本和延
  • Organizer: International Meeting on "Pathogenic mechanisms and emerging therapies for PD and other neurological disorders"
  • Int'l Joint Research / Invited
• [Presentation] Migration of new neurons for maintenance and repair of adult brain. (2017)
  • Author(s): 澤本和延
  • Organizer: コペンハーゲン大学招待講演
  • Invited
• [Presentation] 脳の老化・再生過程における繊毛の変化 (2017)
  • Author(s): 澤本和延
  • Organizer: 第69回日本細胞生物学会
  • Int'l Joint Research / Invited
• [Presentation] 生後の脳内を移動する新生ニューロンの三次元形態解析 (2017)
  • Author(s): 澤本和延, 松本真実, 澤田雅人, 金子奈穂子, 大野伸彦
  • Organizer: 第73回日本顕微鏡学会学術講演会
  • Invited
• [Book] ライフサイエンス新着論文レビュー (2018)
  • Author(s): 神農英雄, 澤本和延
  • Publisher: ライフサイエンス統合データベースセンター
• [Remarks] 澤本和延研究室ホームページ
  • URL: http://k-sawamoto.com/
• [Remarks] 神経化学トピックス 30
  • URL: http://www.neurochemistry.jp/mu1qe6y76-15/#_15
• [Remarks] 名古屋市立大学ホームページ 脳梗塞後の神経再生メカニズムを発見
  • URL: https://www.nagoya-cu.ac.jp/about/press/press/release/files/20181210/301213.pdf
• [Remarks] 生理学研究所ホームページ 脳梗塞後の神経再生メカニズムを発見
  • URL: https://www.nips.ac.jp/release/2018/12/post_381.html
• [Remarks] EurekAlert! 神経細胞が脳内を移動するための仕組みを解明
  • URL: https://www.eurekalert.org/pub_releases_ml/2018-10/nios-j101718.php
• [Remarks] 神経細胞の脳内の位置を決定するしくみ
  • URL: http://www.nagoya-cu.ac.jp/about/press/press/release/files/20180118/300119.pdf
• [Remarks] 神経細胞の脳内の位置を決定するしくみ
  • URL: http://www.nips.ac.jp/release/2018/01/post_355.html
• [Remarks] 新生児期に存在する脳障害後の神経再生メカニズムを発見
  • URL: http://www.nagoya-cu.ac.jp/about/press/press/release/files/20171222/291222.pdf
• [Remarks] 新生児期に存在する脳障害後の神経再生メカニズムを発見
  • URL: http://www.nips.ac.jp/release/2017/12/post_354.html