Molecular anatomy of photoreceptor ribbon synapses with special attention to the ADP ribosylation factor 6 (Arf6) pathway

• Project/Area Number: 15H04672
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General anatomy (including histology/embryology)
• Research Institution: Kitasato University
• Principal Investigator: SAKAGAMI HIROYUKI 北里大学, 医学部, 教授 (90261528)
• Research Collaborator: FUKAYA Masahiro ; HARA Yoshinobu
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000); Fiscal Year 2018: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2017: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2016: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2015: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
• Keywords: リボンシナプス / 視覚 / 筋ジストロフィー / 小胞輸送 / シナプス / 低分子量GTP結合タンパク質 / 網膜 / 光感受細胞 / 視細胞 / 光受容 / 低分子量Gタンパク質 / ジストロフィン

Outline of Final Research Achievements

The aim of this study is to provide the functional importance of the ADP ribosylation factor 6 (Arf6) pathway in the photoreceptor ribbon synapse. By immunofluorescence and immunoelectron microscopy using a novel anti-BRAG2a antibody, we found that BRAG2a, a guanine nucleotide exchange factor specific for Arf6, colocalized to the plasma membrane of lateral walls and processes of photoreceptor terminals within the synaptic cavity with dystroglycan and dystrophin. Pull-down and immunoprecipitation assays using the retinal lysate demonstrated the complex formation of BRAG2a with the dystrophin and beta-dystroglycan. The present study provides evidence for BRAG2a as a novel component of the photoreceptor dystrophin-associated glycoprotein complex (DGC), suggesting the possible functional involvement of the BRAG2a-Arf6 pathway downstream of the DGC in synaptic organization of photoreceptors.

Academic Significance and Societal Importance of the Research Achievements

ヒトの外界からの情報の大半は視覚に依存しており、視覚障害が生活の質(QOL)に与える影響は著しく大きい。網膜の視細胞は、明暗・色調を鋭敏に感知して神経伝達物質を持続的に放出する特殊なシナプス「リボンシナプス」を持つ。しかしながら、リボンシナプスの生理学的特性を支える分子制御機序は未だ不明である。本研究は、低分子量G蛋白質Arf6経路の網膜リボンシナプスでの機能を明らかにするために実施した.
その結果、視細胞リボンシナプスにおいてArf6活性化制御因子BRAG2aがジストロフィン複合体を形成することを見出し、リボンシナプスの形成・維持に関わる新たな制御経路である可能性を示した。

Research Products

• [Journal Article] Involvement of SRF coactivator MKL2 in BDNF-mediated activation of the synaptic activity-responsive element in the Arc gene. (2019)
  • Author(s): Kikuchi K, Ihara D, Fukuchi M, Tanabe H, Ishibashi Y, Tsujii J, Tsuda M, Kaneda M, Sakagami H, Okuno H, Bito H, Yamazaki Y, Ishikawa M, Tabuchi A.
  • Journal Title: Journal of Neurochemistry Volume: 148 Issue: 2 Pages: 204-218
  • DOI: 10.1111/jnc.14596
  • Peer Reviewed / Open Access
• [Journal Article] CMT type 2N disease-associated AARS mutant inhibits neurite growth that can be reversed by valproic acid (2019)
  • Author(s): Tatsumi Yuriko、Matsumoto Naoto、Iibe Noriko、Watanabe Natsumi、Torii Tomohiro、Sango Kazunori、Homma Keiichi、Miyamoto Yuki、Sakagami Hiroyuki、Yamauchi Junji
  • Journal Title: Neuroscience Research Volume: 139 Pages: 69-78
  • DOI: 10.1016/j.neures.2018.09.016
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Hyperactivation of mTORC1 disrupts cellular homeostasis in cerebellar Purkinje cells. (2019)
  • Author(s): Sakai Y, Kassai H, Nakayama H, Fukaya M, Maeda T, Nakao K, Hashimoto K, Sakagami H, Kano M, Aiba A.
  • Journal Title: Scientific Reports Volume: Feb 26;9(1) Issue: 1 Pages: 2799-2799
  • DOI: 10.1038/s41598-019-38730-4
  • Peer Reviewed / Open Access
• [Journal Article] Pallidin is a novel interacting protein for cytohesin-2 and regulates the early endosomal pathway and dendritic formation in neurons (2018)
  • Author(s): Ito Akiko、Fukaya Masahiro、Saegusa Shintaro、Kobayashi Emi、Sugawara Takeyuki、Hara Yoshinobu、Yamauchi Junji、Okamoto Hirotsugu、Sakagami Hiroyuki
  • Journal Title: Journal of Neurochemistry Volume: 147 Issue: 2 Pages: 153-177
  • DOI: 10.1111/jnc.14579
  • Peer Reviewed
• [Journal Article] FIP4/Arfophilin-2 plays overlapping but distinct roles from FIP3/Arfophilin-1 in neuronal migration during cortical layer formation (2018)
  • Author(s): Hara Yoshinobu、Fukaya Masahiro、Sugawara Takeyuki、Sakagami Hiroyuki
  • Journal Title: European Journal of Neuroscience Volume: 48 Issue: 9 Pages: 3082-3096
  • DOI: 10.1111/ejn.14199
  • Peer Reviewed
• [Journal Article] New Features on the Expression and Trafficking of mGluR1 Splice Variants Exposed by Two Novel Mutant Mouse Lines. (2018)
  • Author(s): Rika Naito, Hidetoshi Kassai, Yusuke Sakai, Sabine Schönherr, Masahiro Fukaya, Christoph Schwarzer, Hiroyuki Sakagami, Kazuki Nakao, Atsu Aiba, Francesco Ferraguti
  • Journal Title: Frontiers in Molecular Neuroscience Volume: 11 Pages: 439-439
  • DOI: 10.3389/fnmol.2018.00439
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] BRAG2a, a guanine nucleotide exchange factor for Arf6, is a component of the dystrophin-associated glycoprotein complex at the photoreceptor terminal. (2017)
  • Author(s): Sakagami H, Katsumata O, Hara Y, Sasaoka T, Fukaya M
  • Journal Title: Investigative Ophthalmology and Visual Science (IOVS) Volume: 58 Issue: 9 Pages: 3795-3803
  • DOI: 10.1167/iovs.17-21746
  • Peer Reviewed / Open Access
• [Journal Article] Cellular and subcellular localization of ADP-ribosylation factor 6 in mouse peripheral tissues (2017)
  • Author(s): Katsumata Osamu、Mori Momoko、Sawane Yusuke、Niimura Tomoko、Ito Akiko、Okamoto Hirotsugu、Fukaya Masahiro、Sakagami Hiroyuki
  • Journal Title: Histochemistry and Cell Biology Volume: 148 Issue: 6 Pages: 577-596
  • DOI: 10.1007/s00418-017-1599-8
  • Peer Reviewed / Open Access
• [Journal Article] CDKL5 controls postsynaptic localization of GluN2B-containing NMDA receptors in the hippocampus and regulates seizure susceptibility. (2017)
  • Author(s): Okuda K, Kobayashi S, Fukaya M, Watanabe A, Murakami T, Hagiwara M, Sato T, Ueno H, Ogonuki N, Komano-Inoue S, Manabe H, Yamaguchi M, Ogura A, Asahara H, Sakagami H, Mizuguchi M, Manabe T, Tanaka T
  • Journal Title: Neurobiology of Diseases Volume: 106 Pages: 158-170
  • DOI: 10.1016/j.nbd.2017.07.002
  • Peer Reviewed / Open Access
• [Journal Article] Data on the effect of knockout of cytohesin-1 in myelination-related protein kinase signaling. (2017)
  • Author(s): Ruri Tsuneishi, Naoto Matsumoto, Misa Itaoka, Yuki Urai, Minami Kaneko, Natsumi Watanabe, Shou Takashima, Yoichi Seki, Takako Morimoto, Hiroyuki Sakagami, Yuki Miyamoto, and Junji Yamauchi
  • Journal Title: Data Brief. Volume: 15 Pages: 234-239
  • DOI: 10.1016/j.dib.2017.09.024
  • Peer Reviewed / Open Access
• [Journal Article] Co-localization of endogenous Arf6 and its activator EFA6D in the granular convoluted tubule cells of mouse submandibular glands under normal conditions and when stimulated by isoproterenol, noradrenaline and carbachol. (2017)
  • Author(s): Tachow A, Thoungseabyoun W, Phuapittayalert L, Petcharat K, Sakagami H, Kondo H, Hipkaeo W.
  • Journal Title: Arch Oral Biol. Volume: 82 Pages: 153-159
  • DOI: 10.1016/j.archoralbio.2017.06.007
  • Peer Reviewed / Open Access
• [Journal Article] Distinct subcellular localization of alternative splicing variants of EFA6D, a guanine nucleotide exchange factor for Arf6, in the mouse brain. (2016)
  • Author(s): Fukaya M, Ohata S, Hara Y, Tamaki H, Sakagami H.
  • Journal Title: Journal of Comparative Neurology Volume: 477 Issue: 13 Pages: 868-873
  • DOI: 10.1002/cne.24048
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Expression of β subunit of Ca2+/calmodulin-dependent protein kinase I in the developing rat retina. (2016)
  • Author(s): Jusuf AA, Sakagami H, Kikkawa S, Terashima T.
  • Journal Title: Kobe Journal of Medical Sciences Volume: 61
  • Peer Reviewed
• [Journal Article] Polyhydramnios in Lrp4 knockout mice with bilateral kidney agenesis: Defects in the pathways of amniotic fluid clearance. (2016)
  • Author(s): Tanahashi, H.;Tian, Q.B.; Hara, Y.;Sakagami, H.;Endo, S.;Suzuki, T.
  • Journal Title: Sci Rep Volume: 6 Issue: 1 Pages: 20241-20241
  • DOI: 10.1038/srep20241
  • NAID: 120007100143
  • Peer Reviewed / Open Access
• [Journal Article] Interaction of serologically defined colon cancer antigen-3 with Arf6 and its predominant expression in the mouse testis. (2016)
  • Author(s): Sakagami H, Hara Y, Fukaya M.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 477 Issue: 4 Pages: 868-873
  • DOI: 10.1016/j.bbrc.2016.06.150
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] ADP ribosylation factor 6 regulates neuronal migration in the developing cerebral cortex through FIP3/Arfophilin-1-dependent endosomal trafficking of N-cadherin. (2016)
  • Author(s): Yoshinobu Hara, Masahiro Fukaya, Kanehiro Hayashi, Takeshi Kawauchi, Kazunori Nakajima, and Hiroyuki Sakagami
  • Journal Title: eNeuro Volume: 3 Issue: 4 Pages: 1-20
  • DOI: 10.1523/eneuro.0148-16.2016
  • Peer Reviewed / Open Access
• [Journal Article] Targeted disruption of the mouse protein phosphatase ppm1l gene leads to structural abnormalities in the brain. (2016)
  • Author(s): Kusano, R., Kousuke Fujita, K., Yasuharu Shinoda, Y., Nagaura, Y., Kiyonari, H., Abe, T, Watanabe, T., Matsui, Y., Fukaya, M., Sakagami, H., Sato, T., Funahashi, J.-I., Ohnishi, M., Tamura, S., Kobayashi, T.
  • Journal Title: FEBS Letters Volume: 590 Issue: 20 Pages: 3603-3615
  • DOI: 10.1002/1873-3468.12429
  • Peer Reviewed
• [Journal Article] Arf6 guanine-nucleotide exchange factor cytohesin-2 regulates myelination in nerves. (2015)
  • Author(s): Tomohiro Torii, Nobuhiko Ohno, Yuki Miyamoto, Kazuko Kawahara, Yurika Saitoh, Kazuaki Nakamura, Shou Takashima, Hiroyuki Sakagami, Akito Tanoue, and Junji Yamauchi
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 460 Issue: 3 Pages: 819-825
  • DOI: 10.1016/j.bbrc.2015.03.113
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Arf6 mediates Schwann cell differentiation and myelination. (2015)
  • Author(s): Tomohiro Torii, Yuki Miyamoto, Masahiro Yamamoto, Katsuya Ohbuchi, Hideki Tsumura, Kazuo Kawahara, Akito Tanoue, Hiroyuki Sakagami, and Jyunji Yamauchi
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 465 Issue: 3 Pages: 450-457
  • DOI: 10.1016/j.bbrc.2015.08.038
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Novel splice variants in the 5’UTR of Gtf2i expressed in the rat brain: alternative 5’UTRs and differential expression in the neuronal dendrites. (2015)
  • Author(s): Yoshinori Shirai, Masahiko Watanabe, Hiroyuki Sakagami and Tatsuo Suzuki
  • Journal Title: J. Neurochemistry Volume: 134 Issue: 3 Pages: 578-589
  • DOI: 10.1111/jnc.13136
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Stimulation of Sigma-1 Receptor Ameliorates Depressive-like Behaviors in CaMKIV Null Mice. (2015)
  • Author(s): Moriguchi, S.;Sakagami, H.;Yabuki, Y.;Sasaki, Y.;Izumi, H.;Zhang, C.;Han, F.;Fukunaga, K.
  • Journal Title: Mol Neurobiol Volume: 52 Issue: 3 Pages: 1210-1222
  • DOI: 10.1007/s12035-014-8923-2
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Thromboxane A2 induces blood flow recovery via platelet adhesion to ischaemic regions. (2015)
  • Author(s): Amano H, Ito Y, Eshima K, Kato S, Ogawa F, Hosono K, Oba K, Tamaki H, Sakagami H, Shibuya M, Narumiya S, Majima M.
  • Journal Title: Cardiovasc Res. Volume: 107 Issue: 4 Pages: 509-521
  • DOI: 10.1093/cvr/cvv139
  • Peer Reviewed / Open Access
• [Journal Article] The Role of Vascular Endothelial Growth Factor Receptor-1 Signaling in the Recovery from Ischemia. (2015)
  • Author(s): Amano H, Kato S, Ito Y, Eshima K, Ogawa F, Takahashi R, Sekiguchi K, Tamaki H, Sakagami H, Shibuya M, Majima M.
  • Journal Title: PLoS One Volume: 10(7) Issue: 7 Pages: e0131445-e0131445
  • DOI: 10.1371/journal.pone.0131445
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] 低分子量Gタンパク質Arf6活性化因子EFA6Cの小脳シナプス形成における役割 (2019)
  • Author(s): 三枝信太郎、深谷 昌弘 、阪上 洋行
  • Organizer: 第124回日本解剖学会総会・全国学術集会
• [Presentation] Arf6活性化制御因子cytohesin 2 の局在解析と疼痛制御機構における役割 (2019)
  • Author(s): 伊藤諭子、深谷昌弘、岡本浩嗣、阪上洋行
  • Organizer: 第124回日本解剖学会総会・全国学術集会
• [Presentation] 大脳皮質層形成におけるArf 結合タンパク質Arfophilin-1 と-2 の機能的差異 (2018)
  • Author(s): 原芳伸、阪上洋行
  • Organizer: 第106回日本解剖学会関東支部会
• [Presentation] 脊髄におけるCytohesin 2 の局在解析と疼痛制御の機能関与 (2018)
  • Author(s): 伊藤諭子、深谷昌弘、岡本浩嗣、阪上洋行
  • Organizer: 第106回日本解剖学会関東支部会
• [Presentation] 新規cytohesin-2結合分子Pallidinの初期エンドソーム輸送調節と樹状突起形成制御機構 (2018)
  • Author(s): 深谷 昌弘、伊藤 諭子、三枝信太郎 、原 芳伸、阪上 洋行
  • Organizer: 第123回日本解剖学会総会・全国学術集会(東京)
• [Presentation] 中枢神経系髄鞘形成におけるArf6活性調節因子cytohesin-2の機能 (2017)
  • Author(s): 加倉井俊也, 深谷昌弘, 山内淳司, 阪上洋行
  • Organizer: 第122回日本解剖学会総会・全国学術集会
  • Place of Presentation: 長崎大学坂本キャンパス(長崎県長崎市）
  • Year and Date: 2017-03-28
• [Presentation] 新規cytohesin-2結合分子Pallidinの生体マウス脳における細胞内局在解析 (2017)
  • Author(s): 深谷昌弘, 小林瑛未, 阪上洋行
  • Organizer: 第122回日本解剖学会総会・全国学術集会
  • Place of Presentation: 長崎大学坂本キャンパス(長崎県長崎市）
  • Year and Date: 2017-03-28
• [Presentation] 新規cytohesin-2結合分子Pallidinの細胞内局在とその機能 (2017)
  • Author(s): 深谷 昌弘、伊藤 諭子、岡本 浩嗣、阪上 洋行
  • Organizer: 第40回日本神経科学大会(千葉）
• [Presentation] 神経細胞の移動におけるRab11-FIP3，FIP4の機能解析 (2016)
  • Author(s): 原 芳伸, 阪上 洋行
  • Organizer: 第39回日本神経科学大会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-07-20
• [Presentation] Arf6活性制御因子BRAG2cはジストロフィン複合体との相互作用を介して網膜視細胞のシナプスに局在する (2016)
  • Author(s): 阪上 洋行, 深谷 昌弘
  • Organizer: 第39回日本神経科学大会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-07-20
• [Presentation] シナプス後部におけるBRAG2-Arf6シグナル経路を介したAMPA受容体輸送調節機構 (2016)
  • Author(s): 深谷 昌弘, 阪上 洋行
  • Organizer: 第121回日本解剖学会総会・全国学術集会
  • Place of Presentation: ビックパレットふくしま（郡山市）
  • Year and Date: 2016-03-30
• [Presentation] BRAG2-Arf6経路による新たなAMPA受容体シナプス発現調節機構 (2015)
  • Author(s): 阪上 洋行
  • Organizer: BMB2015(第86回日本生化学会大会・第38回日本分子生物学会）
  • Place of Presentation: 神戸国際会議場 (神戸市中央区）
  • Year and Date: 2015-11-07
• [Presentation] BRAG2はendophilin3との相互作用を介してシナプス後部でのArf6の活性化とAMPA型受容体のエンドサイトーシスを調節する (2015)
  • Author(s): 深谷 昌弘, 阪上 洋行
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸国際会議場 (神戸市中央区）
  • Year and Date: 2015-07-28
• [Presentation] ADPリボシル化因子6 (Arf6)は脳層形成において神経細胞の移動を制御する (2015)
  • Author(s): 原 芳伸, 阪上 洋行
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸国際会議場 (神戸市中央区）
  • Year and Date: 2015-07-28
• [Presentation] グリシン作動性ニューロンでCreを発現するマウス系統の開発 (2015)
  • Author(s): 柿崎 利和, 阪上 洋行, 崎村 建司, 柳川 右千夫
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸国際会議場 (神戸市中央区）
  • Year and Date: 2015-07-28
• [Remarks] 北里大学医学部解剖学（阪上）単位ホームページ
  • URL: http://www.med.kitasato-u.ac.jp/~sakagami/
• [Remarks] 北里大学医学部解剖学(阪上）単位ホームページ
  • URL: http://www.med.kitasato-u.ac.jp/~sakagami/
• [Remarks] 北里大学医学部解剖学（阪上)単位ホームページ
  • URL: http://www.med.kitasato-u.ac.jp/~sakagami/index.php?FrontPage
• [Remarks] 北里大学医学部解剖学(阪上)単位ホームページ
  • URL: http://www.med.kitasato-u.ac.jp/~sakagami/index.php?FrontPage