Study on the molecular mechanisms underlying CAPS-mediated exocytosis of dense-core vesicles containing BDNF and catecholamine

• Project/Area Number: 23300137
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Neurochemistry/Neuropharmacology
• Research Institution: Tokyo University of Science
• Principal Investigator: FURUICHI Teiichi 東京理科大学, 理工学部, 教授 (50219094)
• Project Period (FY): 2011-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥20,150,000 (Direct Cost: ¥15,500,000、Indirect Cost: ¥4,650,000); Fiscal Year 2013: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2012: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2011: ¥8,970,000 (Direct Cost: ¥6,900,000、Indirect Cost: ¥2,070,000)
• Keywords: CAPS1 / CAPS2 / 有芯小胞 / BDNF / トランスゴルジ網 / 自閉症 / CAPS2-dex3 / NT-3 / 社会行動 / Cadps2 / Arf4/5 / ゴルジ体 / 発達障害

Research Abstract

We showed that CAPS1 and CAPS2 interacts with Arf4/5, which is possibly involved in dense-core vesicle (DCV) trafficking in the trans-Golgi network (TGN). Dex3 (deletion of exon3) is a rare alternative splicing CAPS2 variant which was identified to be increased in some of patients with autism. We generated a dex3-expressing mice and verified an impairment in axonal dex3 localization, contributing to a reduction in BDNF release from axons and thereby resulting in abnormal synapses. Dex3 mice also showed deficits in social and anxiety behaviors, suggesting a possible association of dex3 with brain development and behavior related to autism. Although CAPS1 regulates insulin secretion, its function in the brain is essentially unknown because of neonatal death of CAPS1 KO mice. We generated CAPS1 conditional knockout mice and suggested that loss of CAPS1 disrupts the TGN-DCV pathway and BDNF release, which may be associated with a mechanism underlying diabetes and comorbid depression.

Research Products

• [Journal Article] CAPS1 deficiency perturbs dense-core vesicle trafficking and Golgi structure and reduces presynaptic release probability in the mouse brain (2013)
  • Author(s): Sadakata T, Kakegawa W, Shinoda Y, Hosono M, Katoh-SembaR, Sekine Y, Sato Y, Tanaka M, Iwasato T, Itohara S, Furuyama K, Kawaguchi Y, Ishizaki Y, Yuzaki M and Furuichi T
  • Journal Title: J. Neurosci Volume: 33 Pages: 17326-17334
• [Journal Article] Activation of cultured astrocytes by amphotericin B : Stimulation of NO and cytokines production and changes in neurotrophic factors (2013)
  • Author(s): Motoyoshi-Yamashiro A, Tamura M, Moriyama M, Takano K, Kawabe K, Nakajima H, Katoh-Semba R, Furuichi T and Nakamura Y
  • Journal Title: Neurochem. Int Volume: 63 Pages: 93-100
• [Journal Article] Astrocytic Ca^<2+> signals are required for the functional integrity of tripartite synapses (2013)
  • Author(s): Tanaka, M., Shih, PY., Gomi, H., Yoshida, H., Nakai, J., Ando, R., Furuichi, T., Mikoshiba, K., Semyanov, A., and Itohara, S
  • Journal Title: Mol. Brain Volume: 6 Pages: 6-6
• [Journal Article] Autistic-like behavioral phenotypes in a mouse model with copy number variation of the CAPS2/CADPS2 gene (2013)
  • Author(s): Sadakata, T., Shinoda, Y., Oka, M., Sekine, Y., and Furuichi, T
  • Journal Title: FEBS Lett Volume: 587 Pages: 54-59
• [Journal Article] Animal Models of Autism Spectrum Disorder (ASD) : A Synaptic-level Approach to Autistic-like Behavior in Mice (2013)
  • Author(s): Shinoda, Y., Sadakata, T., and Furuichi, T
  • Journal Title: Exp. Anim Volume: 62(2) Pages: 71-78
  • NAID: 10031163545
• [Journal Article] Mouse models of mutations and variations in autism spectrum disorder-associated genes : mice expressing caps2/cadps2 copy number and alternative splicing variants (2013)
  • Author(s): Sadakata, T., Shinoda, Y., Sato, A., Iguchi, H., Ishii, C., Matsuo, M., Yamaga, R., Furuichi, T
  • Journal Title: Int J Environ Res Public Health Volume: 10(12) Pages: 6335-6353
• [Journal Article] CAPS1 deficiency perturbs dense-core vesicle trafficking and Golgi structure and reduces presynaptic release probability in the mouse brain (2013)
  • Author(s): Sadakata T, Kakegawa W, Shinoda Y, Hosono M, Katoh-Semba R, Sekine Y, Sato Y, Tanaka M, Iwasato T, Itohara S, Furuyama K, Kawaguchi Y, Ishizaki Y, Yuzaki M, Furuichi T.
  • Journal Title: Journal of Neuroscience. Volume: 33 Pages: 17326-17334
  • DOI: 10.1523/jneurosci.2777-13.2013
  • Peer Reviewed
• [Journal Article] Autistic-like behavioral phenotypes in a mouse model with copy number variation of the CAPS2/CADPS2 gene. (2013)
  • Author(s): Sadakata T, Shinoda Y, Oka M, Sekine Y, Furuichi T.
  • Journal Title: FEBS Letter. Volume: 587 Pages: 54-59
  • DOI: 10.1016/j.febslet.2012.10.047
  • Peer Reviewed
• [Journal Article] Mouse models of mutations and variations in autism spectrum disorder-associated genes: mice expressing Caps2/Cadps2 copy number and alternative splicing variants. (2013)
  • Author(s): Sadakata T, Shinoda Y, Sato A, Iguchi H, Ishii C, Matsuo M, Yamaga R, Furuichi T
  • Journal Title: International Journal of Environmental Research and Public Health. Volume: 10 Pages: 6335-53
  • DOI: 10.3390/ijerph10126335
  • Peer Reviewed
• [Journal Article] Animal Models of Autism Spectrum Disorder (ASD): A Synaptic-Level Approach to Autistic-Like Behavior in Mice (2013)
  • Author(s): Shinoda Y, Sadakata T, Furuichi T
  • Journal Title: Exp. Anim. Volume: 62 Issue: 2 Pages: 71-78
  • DOI: 10.1538/expanim.62.71
  • NAID: 10031163545
  • Peer Reviewed
• [Journal Article] Reduced axonal localization of a Caps2 splice variant impairs axonal release of BDNF and causes autistic-like behavior in mice (2012)
  • Author(s): Sadakata, T., Shinoda, Y., Oka, M., Sekine, Y., Sato, Y., Saruta, C., Miwa, H., Tanaka, M., Itohara, S., and Furuichi, T
  • Journal Title: Proc. Natl. Acad. Sci. U.S.A Volume: 109 Pages: 21104-21109
• [Journal Article] Clustered fine compartmentalization of the mouse embryonic cerebellar cortex is rearranged into the postnatal striped organization (2012)
  • Author(s): Fujita, H., Morita, N., Unno, T., Furuichi, T., and Sugihara, I
  • Journal Title: J. Neurosci Volume: 32 Pages: 15688-15703
• [Journal Article] Calcium-dependent activator protein for secretion 2 interacts with the class II ARF small GTPases and regulates dense-core vesicle trafficking (2012)
  • Author(s): Sadakata, T., Sekine, Y., Oka, M., Itakura, M., and Furuichi, T
  • Journal Title: FEBS J. Volume: 279 Pages: 384-394
• [Journal Article] Reduced axonal localization of a Caps2 splice variant impairs axonal release of BDNF and causes autistic-like behavior in mice. (2012)
  • Author(s): Sadakata, T., Shinoda, Y., Oka, M., Sekine, Y., Sato, Y., Saruta, C., Miwa, H., Tanaka, M., Itohara, S., and Furuichi, T.
  • Journal Title: Proc Natl Acad Sci U S A. Volume: 109 Pages: 21104-21109
  • DOI: 10.1073/pnas.1210055109
  • Peer Reviewed
• [Journal Article] Clustered Fine Compartmentalization of the Mouse Embryonic Cerebellar Cortex and Its Rearrangement into the Postnatal Striped Configuration. (2012)
  • Author(s): Fujita H, Morita N, Furuichi T, Sugihara I
  • Journal Title: The Journal of Neuroscience Volume: 32 Pages: 15688-15703
  • DOI: 10.1523/jneurosci.1710-12.2012
  • Peer Reviewed
• [Journal Article] BDNFの分泌制御と脳神経回路の形成～CAPS2によるBDNF分泌促進とその機能的役割～ (2012)
  • Author(s): 篠田陽
  • Journal Title: 生化学 Volume: 84 Pages: 106-111
  • NAID: 10030346386
  • Peer Reviewed
• [Journal Article] Calcium-dependent activator protein for secretion 2 interacts with the class II ARF small GTPases and regulates dense-core vesicle trafficking. (2012)
  • Author(s): Sadakata T
  • Journal Title: FEBS JOURNAL Volume: 279 Pages: 384-394
  • DOI: 10.1111/j.1742-4658.2011.08431.x
  • Peer Reviewed
• [Journal Article] 自閉症の発症要因と動物モデル (2012)
  • Author(s): 篠田陽
  • Journal Title: 実験医学増刊号 Volume: 30 Pages: 236-241
  • Peer Reviewed
• [Journal Article] PLD4 is involved in phagocytosis of microglia : expression and localization changes of PLD4 are correlated with activation state of micrglia (2011)
  • Author(s): Otani, Y., Yamaguchi, Y., Sato, Y., Furuichi, T., Ikenaka, K., Kitani, H., and Baba, H
  • Journal Title: PLoS One Volume: 6(11)
• [Journal Article] Phase advance of the light-dark cycle perturbs diurnal rhythms of brain-derived neurotrophic factor and neurotrophin-3 protein levels, which reduces synaptophysin-positive presynaptic terminals in the cortex of juvenile rats (2011)
  • Author(s): Hamatake, M., Miyazaki, N., Sudo, K., Matsuda, M., Sadakata, T., Furuya, A., Ichisaka, S., Hata, Y., Nakagawa, C., Nagata, K., Furuichi, T., Katoh-Semba, R
  • Journal Title: J. Biol. Chem Volume: 286 Pages: 21478-21487
• [Journal Article] Interaction between very-KIND RasGEF and MAP2, and its role in dendrite growth : structure and function of the second kinase non-catalytic C-lobe domain (2011)
  • Author(s): Huang, J., Furuya, A., Hayashi, K., and Furuichi, T
  • Journal Title: FEBS J Volume: 278 Pages: 1651-1661
• [Journal Article] Calcium-dependent activator protein for secretion 2 (CAPS2) promotes BDNF secretion and is critical for the development of GABAergic interneuron network (2011)
  • Author(s): Shinoda, Y., Sadakata, T., Nakao, K., Katoh-Semba, R., Kinameri, E., Furuya, A., Yanagawa, Y., Hirase, H., and Furuichi, T
  • Journal Title: Proc. Natl. Acad. Sci. U.S.A Volume: 108 Pages: 373-378
  • NAID: 120002771000
• [Journal Article] Systematizing and cloning of genes involved in the cerebellar cortex circuit development (2011)
  • Author(s): Furuichi, T., Shiraishi-Yamaguchi, Y., Sato, A., Sadakata, T., Huang, J., Shinoda, Y., Hayashi, K., Mishima, Y., Tomomura, M., Nishibe, H., and Yoshikawa, F
  • Journal Title: Neurochem Res Volume: 36 Pages: 1241-1252
• [Journal Article] Interaction between very-KIND RasGEF and MAP2, and its role in dendrite growth: structure and function of the second kinase non-catalytic C-lobe domain. (2011)
  • Author(s): Huang J
  • Journal Title: FEBS JOURNAL Volume: 278 Pages: 1651-1661
  • DOI: 10.1111/j.1742-4658.2011.08085.x
  • Peer Reviewed
• [Journal Article] Phase advance of the light dark cycle perturbs diurnal rhythms of brain derived neurotrophic factor and neurotrophin 3 levels, which reduces synaptophysin positive presynaptic terminals in the cortex of juvenile rats. (2011)
  • Author(s): Hamatake M, Miyazaki N, Sudo K, Matsuda M, Sadakata T, Furuya A, Ichisaka S, Hata Y, Nakagawa C, Nagata K, Furuichi T, Katoh-Semba R
  • Journal Title: JOURNAL OF BIOLOGICAL CHEMISTRY Volume: 286 Pages: 21478-21487
  • DOI: 10.1074/jbc.m110.195859
  • Peer Reviewed
• [Journal Article] Systematizing and cloning of genes involved in the cerebellar cortex circuit development. (2011)
  • Author(s): Teiichi Furuichi, Yoko Shiraishi-Yamaguchi, Akira Sato, Tetsushi Sadakata, Jinhong Huang, Yo Shinoda, Kanehiro Hayashi, Yuriko Mishima, Mineko Tomomura, Hirozumi Nishibe, Fumio Yoshikawa
  • Journal Title: NEUROCHEMICAL RESEARCH Volume: 36(7) Pages: 1241-1252
  • DOI: 10.1007/s11064-011-0398-1
  • Peer Reviewed
• [Journal Article] PLD4 Is Involved in Phagocytosis of Microglia: Expression and Localization Changes of PLD4 Are Correlated with Activation State of Microglia (2011)
  • Author(s): Otani O
  • Journal Title: PLOS ONE Volume: 6 Pages: 27544-27544
  • DOI: 10.1371/journal.pone.0027544
  • Peer Reviewed
• [Journal Article] 自閉症感受性候補遺伝子CAPS2/CADPS2における遺伝的多型とノックアウトマウスの行動障害 (2011)
  • Author(s): 定方哲史
  • Journal Title: 医学のあゆみ／精神発達遅延・自閉症の分子医学 Volume: 239 Pages: 701-707
  • Peer Reviewed
• [Presentation] 脳の正常な発達とその障害-分泌関連因子CAPS ファミリーの機能 (2013)
  • Author(s): 古市貞一
  • Organizer: 都医学研セミナー
  • Year and Date: 2013-01-11
• [Presentation] Ca^<2+>依存性分泌関連タンパク質CAPS ファミリーのマウス胚脳神経系における遺伝子発現 (2013)
  • Author(s): 三島百合子, 須田翔子, 猿田千尋, 定方哲史, 篠田陽, 和田直之, 小島正巳, 若菜茂晴, 古市貞一
  • Organizer: 第86回日本生化学会大会
• [Presentation] CAPS1 コンディショナル・ノックアウトマウスにおけるシナプス伝達 (2013)
  • Author(s): 石井千晶, 篠田陽, 定方哲史, 古市貞一
  • Organizer: Neuro2013
• [Presentation] マウスドーパミン作動性神経の初代培養系における大型有芯小胞からのドーパミン分泌とCAPS2 の関係 (2013)
  • Author(s): 井口大壽, 小林翔太, 定方哲史, 篠田陽, 古市貞一
  • Organizer: Neuro2013
• [Presentation] Ca2+依存性分泌関連タンパク質CAPSファミリーのマウス胚脳神経系における遺伝子発現 (2013)
  • Author(s): 三島百合子、須田翔子、猿田千尋、定方哲史、篠田陽、和田直之、小島正巳、若菜茂晴、古市貞一
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜
• [Presentation] ナプトタグミン３：ポストシナプスタンパク質の表面発現制御因子 (2013)
  • Author(s): 篠田陽、阿部麗実、古市貞一
  • Organizer: Neuro2013
  • Place of Presentation: 京都・国立京都国際会館
• [Presentation] CAPS1コンディショナル・ノックアウトマウスにおけるシナプス伝達 (2013)
  • Author(s): 石井千晶、篠田陽、定方哲史、古市貞一
  • Organizer: Neuro2013
  • Place of Presentation: 京都・国立京都国際会館
• [Presentation] マウスドーパミン作動性神経の初代培養系における大型有芯小胞からのドーパミン分泌とCAPS2の関係 (2013)
  • Author(s): 井口大壽、小林翔太、定方哲史、篠田陽、古市貞一
  • Organizer: Neuro2013
  • Place of Presentation: 京都・国立京都国際会館
• [Presentation] 脳の正常な発達とその障害-分泌関連因子CAPSファミリーの機能 (2013)
  • Author(s): 古市貞一
  • Organizer: 都医学研セミナー
  • Place of Presentation: 東京都医学総合研究所
  • Invited
• [Presentation] 胎児期低栄養曝露された申請仔の遺伝子発現 (2013)
  • Author(s): 若菜茂晴、古瀬民生、幸田尚、古市貞一、三宅邦夫、久保田建夫
  • Organizer: 第83回日本衛生学会学術総会
  • Place of Presentation: 金沢大学鶴間・宝町キャンパス
• [Presentation] マウス初代培養ドーパミンニューロンにおけるCAPSタンパク質の発現と大型有芯小胞開口放出との関係 (2013)
  • Author(s): 井口大壽、小林翔太、定方哲史、篠田陽、古市貞一
  • Organizer: 第90回日本生理学会大会
  • Place of Presentation: 東京
• [Presentation] CADPSタンパク質は視床下部オキシトシン産生ニューロンで発現し、下垂体後葉と共局在する (2013)
  • Author(s): 山鹿亮祐、長江彩那、定方哲史、篠田陽、古市貞一
  • Organizer: 第90回日本生理学会大会
  • Place of Presentation: 東京
• [Presentation] CAPS2によるBDNFの放出の制御 (2013)
  • Author(s): 松尾誠、中川直紀、篠田陽、古市貞一
  • Organizer: 第90回日本生理学会大会
  • Place of Presentation: 東京
• [Presentation] BDNFによって増強される興奮性シナプス小胞分泌のメカニズム (2013)
  • Author(s): 土井樹、古市貞一、篠田陽、Camin Dean
  • Organizer: 第90回日本生理学会大会
  • Place of Presentation: 東京
• [Presentation] 刺激の違いによるBDNF放出動態の比較とCAPS2活性に関わるCa2+チャネルの特定 (2013)
  • Author(s): 中川直紀、篠田陽、古市貞一、松尾誠
  • Organizer: 第90回日本生理学会大会
  • Place of Presentation: 東京
• [Presentation] Synaptotagmin3：小胞分泌－取り込み制御分子としての役割 (2013)
  • Author(s): 阿部麗実、篠田陽、古市貞一、Camin Dean
  • Organizer: 第90回日本生理学会大会
  • Place of Presentation: 東京
• [Presentation] CAPS1 cKO マウスの海馬におけるシナプス伝達 (2013)
  • Author(s): 石井千晶、篠田陽、定方哲史、古市貞一
  • Organizer: 第90回日本生理学会大会
  • Place of Presentation: 東京
• [Presentation] PC12におけるVMAT2-pHluorin利用した大型有芯小胞の開口放出イメージング (2013)
  • Author(s): 小林翔太、佐藤友美、篠田陽、定方哲史、井口大壽、古市貞一
  • Organizer: 第90回日本生理学会大会
  • Place of Presentation: 東京
• [Presentation] 脳特異的BAD-LAMPの発現と機能 (2013)
  • Author(s): 齊藤由弦、藤原真奈、篠田陽、古市貞一
  • Organizer: 第90回日本生理学会大会
  • Place of Presentation: 東京
• [Presentation] 脳の正常な発達とその障害-分泌関連因子CAPS2 の役割について (2012)
  • Author(s): 古市貞一
  • Organizer: 埼玉大学・研究機構 脳科学融合研究センター シンポジウム(III) 脳の発生・発達とその破綻の理解をめざして
  • Year and Date: 2012-10-06
• [Presentation] Co-localization of Ca^<2+>-dependent activator protein for secretion (CAPS) proteins with oxytocin in the mouse pituitary (2012)
  • Author(s): Ayana Nagae, Yumi Sato, Tetsushi Sadakata, Yo Shinoda, and Teiichi Furuichi
  • Organizer: 第35回日本神経科学大会
• [Presentation] 有芯小胞分泌調節分子CAPS2 遺伝子欠損マウスにおける行動障害 (2012)
  • Author(s): 古市貞一
  • Organizer: 日本実験動物科学・技術
  • Place of Presentation: 九州
• [Presentation] 有芯小胞分泌調節分子CAPS2遺伝子欠損マウスにおける行動障害 (2012)
  • Author(s): 古市貞一
  • Organizer: 日本実験動物科学・技術 九州 2012
  • Place of Presentation: 別府、大分県
  • Invited
• [Presentation] 脳の正常な発達とその障害-分泌関連因子CAPS2の役割について (2012)
  • Author(s): 古市貞一
  • Organizer: 埼玉大学・研究機構 脳科学融合研究センター シンポジウム（III） 脳の発生・発達とその破綻の理解をめざして
  • Place of Presentation: 埼玉大学、埼玉県
  • Invited
• [Presentation] Co-localization of Ca2+-dependent activator protein for secretion (CAPS)proteins with oxytocin in the mouse pituitary (2012)
  • Author(s): Ayana Nagae, Yumi Sato, Tetsushi Sadakata, Yo Shinoda, and Teiichi Furuichi
  • Organizer: 第３５回日本神経科学大会
  • Place of Presentation: 名古屋市国際会議場
• [Presentation] Examining the effects of chronic corticosterone administration in Ca^<2+>-dependent activator protein for secretion 2 (Caps2) knockout mice : implication for anxiety/depression-like behaviors (2011)
  • Author(s): Mishima Y, Sadakata T, Semba-Katoh R, Saruta C and Furuichi T
  • Organizer: Annual Meeting of Society for Neuroscience
• [Presentation] Enhanced secretion of BDNF by CAPS2 is critical for proper brain development and behavior (2011)
  • Author(s): Furuichi T, Sadakata T and Shinoda Y
  • Organizer: The 32nd Naito Conference
• [Presentation] 長期的なコルチコステロン投与による不安・欝様行動への影響 : Ca^<2+>-dependent activator protein for secretion 2(Caps2)遺伝子欠損マウスを用いた実験. (2011)
  • Author(s): 三島百合子, 定方哲史, 仙波りつ子, 猿田千尋, 古市貞一
  • Organizer: 第34回日本神経科学大会
• [Presentation] 長期的なコルチコステロン投与による不安・欝様行動への影響：Ca2+-dependent activator protein for secretion 2（Caps2）遺伝子欠損マウスを用いた実験. (2011)
  • Author(s): 三島百合子
  • Organizer: 第34回日本神経科学大会
  • Place of Presentation: 横浜
• [Presentation] ミクログリアの活性化に伴うPLD4発現調節機構 (2011)
  • Author(s): 大谷嘉典
  • Organizer: 第54回日本神経化学会大会
  • Place of Presentation: 石川
• [Presentation] Enhanced secretion of BDNF by CAPS2 is critical for proper brain development and behavior. (2011)
  • Author(s): Furuichi T
  • Organizer: The 32nd Naito Conference
  • Place of Presentation: yamanashi, Japan
  • Invited
• [Presentation] Examining the effects of chronic corticosterone administration in Ca2+-dependent activator protein for secretion 2 (Caps2) knockout mice: implication for anxiety/depression-like behaviors. (2011)
  • Author(s): Mishima Y
  • Organizer: Annual Meeting of Society for Neuroscience
  • Place of Presentation: Washington DC, USA
• [Book] ゴルジ体における有芯小胞の生合成とCAPSタンパク質の役割,生体の科学 (2012)
  • Author(s): 定方哲史, 篠田 陽, 古市貞一
• [Book] 自閉症の発症要因と動物モデル 実験医学 (2012)
  • Author(s): 篠田 陽, 定方哲史, 三島百合子, 古市貞一
• [Book] BDNF の分泌制御と脳神経回路の形成～CAPS2 によるBDNF 分泌促進とその機能的役割～生化学 (2012)
  • Author(s): 篠田 陽, 古市貞一
• [Book] 自閉症感受性候補遺伝子CAPS2/CADPS2 における遺伝的多型とノックアウトマウスの行動障害 医学のあゆみ (2011)
  • Author(s): 定方哲史, 古市貞一
• [Remarks]
  • URL: http://www.lmn.bs.noda.tus.ac.jp/#!homej/c1lby
• [Remarks] CAPS1 cKOマウスの表現型の解析 糖尿病 とうつ病が併発するメカニズム との関連性を示唆
  • URL: http://www.lmn.bs.noda.tus.ac.jp/#!homej/c1lby
• [Remarks] CAPS2 dex3マウスの表現型の解析 自閉症モデルマウスの作製に成功
  • URL: http://www.lmn.bs.noda.tus.ac.jp/#!homej/c1lby