Elucidation of the molecular mechanism of the paracellular barrier at triellular junctions

• Project/Area Number: 18H02440
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 44010:Cell biology-related
• Research Institution: National Institute for Physiological Sciences
• Principal Investigator: Furuse Mikio 生理学研究所, 生体機能調節研究領域, 教授 (90281089)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000); Fiscal Year 2020: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2019: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2018: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
• Keywords: タイトジャンクション / トリセルラータイトジャンクション / ３細胞結合 / アンギュリン / トリセルリン / 上皮バリア機能 / 3細胞結合 / クローディン / 細胞間接着 / アンギュリンファミリー

Outline of Final Research Achievements

Tricellular tight junctions (tTJs) are intercellular junctions that seal the intercellular space at tricellular contacts, where the vertices of three epithelial cells meet, and contribute to the epithelial barrier function. Tricellulin and angulin family of membrane proteins are known constituents of tTJs, but the roles of these proteins in tTJ formation remains elusive. In this study, we have demonstrated that angulin-1, not tricellulin, is responsible for the plasma membrane contact at tricellular contacts.

Academic Significance and Societal Importance of the Research Achievements

私たちの体の最も基本的は防御機能である上皮バリア機能がはたらくためには、上皮細胞同士の隙間における物質の漏れを防ぐ必要がある。上皮の細胞間隙には、隣り合う２細胞間の隙間と、３つの上皮細胞の角に挟まれる点状の３細胞間の隙間が存在する。前者をシールする細胞間結合タイトジャンクションについては多くの知見の蓄積があるのに対して、後者を塞ぐ細胞間結合トリセルラータイトジャンクションの仕組みについては不明な点が多かった。本研究はトリセルラータイトジャンクション形成の分子機構をこれまでになく詳細かつ明快に示したといえる。上皮バリア機能に関わる本研究は、医学分野への波及も期待される。

Research Products

• [Journal Article] Recent advances in understanding tight junctions (2021)
  • Author(s): Furuse Mikio、Takai Yoshimi
  • Journal Title: Faculty Reviews Volume: 10 Pages: 18-18
  • DOI: 10.12703/r/10-18
  • Peer Reviewed
• [Journal Article] The novel membrane protein Hoka regulates septate junction organization and stem cell homeostasis in the Drosophila gut (2021)
  • Author(s): Izumi Yasushi、Furuse Kyoko、Furuse Mikio
  • Journal Title: Journal of Cell Science Volume: 134 Issue: 6 Pages: 257022-257022
  • DOI: 10.1242/jcs.257022
  • Peer Reviewed
• [Journal Article] The septate junction protein Tetraspanin 2A is critical to the structure and function of Malpighian tubules in Drosophila melanogaster. (2020)
  • Author(s): Klaus W Beyenbach, Frederike Schone, Leonhard F Breitsprecher, Felix Tiburcy, Mikio Furuse, Yasushi Izumi, Heiko Meyer, Sima Jonusaite, Aylin R Rodan, Achim Paululat
  • Journal Title: American journal of physiology. Cell physiology Volume: 318 Issue: 6 Pages: C1107-C1122
  • DOI: 10.1152/ajpcell.00061.2020
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Angulin-2/ILDR1, a tricellular tight junction protein, does not affect water transport in the mouse large intestine (2020)
  • Author(s): Hempstock Wendy、Sugioka Shiori、Ishizuka Noriko、Sugawara Taichi、Furuse Mikio、Hayashi Hisayoshi
  • Journal Title: Scientific Reports Volume: 10 Issue: 1 Pages: 10374-10374
  • DOI: 10.1038/s41598-020-67319-5
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Physiological functions of junctional adhesion molecules (JAMs) in tight junctions (2020)
  • Author(s): Hartmann Christian、Schwietzer Ysabel Alessa、Otani Tetsuhisa、Furuse Mikio、Ebnet Klaus
  • Journal Title: Biochimica et Biophysica Acta (BBA) - Biomembranes Volume: 1862 Issue: 9 Pages: 183299-183299
  • DOI: 10.1016/j.bbamem.2020.183299
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Tight Junction Structure and Function Revisited (2020)
  • Author(s): Otani Tetsuhisa、Furuse Mikio
  • Journal Title: Trends in Cell Biology Volume: 30 Issue: 10 Pages: 805-817
  • DOI: 10.1016/j.tcb.2020.08.004
  • Peer Reviewed
• [Journal Article] The septate junction protein Mesh is required for epithelial morphogenesis, ion transport, and paracellular permeability in the Drosophila Malpighian tubule. (2020)
  • Author(s): Sima Jonusaite, Klaus W. Beyenbach, Heiko Meyer, Achim Paululat, Yasushi Izumi, Mikio Furuse, and Aylin R. Rodan
  • Journal Title: American Journal of Physiology-Cell Physiology Volume: 318 Issue: 3 Pages: C675-C694
  • DOI: 10.1152/ajpcell.00492.2019
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] The extracellular domain of angulin-1 and palmitoylation of its cytoplasmic region are required for angulin-1 assembly at tricellular contacts (2020)
  • Author(s): Yukako Oda, Taichi Sugawara, Yuko Fukata, Yasushi Izumi, Tetsuhisa Otani, Tomohito Higashi, Masaki Fukata, Mikio Furuse
  • Journal Title: Journal of Biological Chemistry Volume: 295 Issue: 13 Pages: 4289-4302
  • DOI: 10.1074/jbc.ra119.010491
  • Peer Reviewed / Open Access
• [Journal Article] Claudins and JAM-A coordinately regulate tight junction formation and epithelial polarity (2019)
  • Author(s): Tetsuhisa Otani, Thanh Phuong Nguyen, Shinsaku Tokuda, Kei Sugihara, Taichi Sugawara, Kyoko Furuse, Takashi Miura, Klaus Ebnet, Mikio Furuse
  • Journal Title: Journal of Cell Biology Volume: 218 Issue: 10 Pages: 3372-3396
  • DOI: 10.1083/jcb.201812157
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Septate junctions regulate gut homeostasis through regulation of stem cell proliferation and enterocyte behavior in Drosophila. (2019)
  • Author(s): Yasushi Izumi, Kyoko Furuse and Mikio Furuse
  • Journal Title: Journal of Cell Science Volume: 132 Issue: 18
  • DOI: 10.1242/jcs.232108
  • Peer Reviewed
• [Journal Article] Claudin-3-deficient C57BL/6J mice display intact brain barriers (2018)
  • Author(s): Castro DM、Coisne C、Lazarevic I、Baden P、Hata M、Iwamoto N、Francisco DMF、Vanlandewijck Mi、He L、Baier FA.、Stroka D、Bruggmann R、Lyck R、Enzmann G、Deutsch U、Betsholtz C、Furuse M、Tsukita Sh、Engelhardt B
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 36731-3
  • DOI: 10.1038/s41598-018-36731-3
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 高分子に対する透過性バリアの形成機構 (2020)
  • Author(s): 大谷哲久, Phuong Nguyen Thanh, 菅原太一, 古瀬幹夫
  • Organizer: 第72回日本細胞生物学会大会
• [Presentation] Angulin-1 regulates the plasme membrane sealing and epithelial barrier function at tricellular contacts independent of tricellulin and claudins (2020)
  • Author(s): 菅原太一, 古瀬京子, 古瀬幹夫
  • Organizer: 第72回日本細胞生物学会大会
• [Presentation] アンギュリンはトリセルリンに依存せず3細胞結合の細胞膜密着構造を形成する (2020)
  • Author(s): 菅原太一, 古瀬京子, 大谷哲久, 若山友彦, 古瀬幹夫
  • Organizer: 第43回日本分子生物学会年会
• [Presentation] 上皮のトリセルラーコンタクトにおけるバリア形成機構 (2020)
  • Author(s): 菅原太一, 古瀬京子, 大谷哲久, 若山友彦, 古瀬幹夫
  • Organizer: 第52回臨床分子形態学会総会・学術集会
• [Presentation] 上皮細胞の隙間をシールする分子機構 (2020)
  • Author(s): 古瀬幹夫
  • Organizer: 第30回日本耳科学会総会・学術講演会
  • Invited
• [Presentation] Epithelial barrier homeostasis by cell competition. (2019)
  • Author(s): Otani T, Furuse M.
  • Organizer: 52回日本発生生物学会大会
• [Presentation] 多様な膜貫通タンパク質により構成されるショウジョウバエのセプテートジャンクション (2019)
  • Author(s): 泉 裕士、古瀬幹夫
  • Organizer: 第19回日本蛋白質化学会年会・第71回日本細胞生物学会大会合同年次大会
• [Presentation] Genetic dissection of tight junctions (2019)
  • Author(s): Otani T, Nguyen TP, Furuse M.
  • Organizer: 第19回日本蛋白質科学会年会・第71回日本発生生物学会大会合同年次大会
• [Presentation] Assembly of angulin-1/LSR in vertices of three epithelial cells is determined by the cytoplasmic palmitoylation. (2019)
  • Author(s): Sugawara T, Oda Y, Fukata Y, Higashi T, Fukata M, Furuse M.
  • Organizer: 第19回日本蛋白質科学会年会・第71回日本発生生物学会大会合同年次大会
• [Presentation] Establishment of a tight junction-deficient epithelial cell line by genome editing of claudin genes (2019)
  • Author(s): Mikio Furuse, Tetsuaki Otani, Taichi Sugawara, Shinsaku Tokuda, Mika Watanabe, Kyoko Furuse, Osamu Nagata
  • Organizer: 9th Federation of the Asian and Oceanian Physiological Sciences
  • Int'l Joint Research
• [Presentation] ZO family proteins regulate epithelial polarity independent of tight junction strand assembly (2019)
  • Author(s): Tetsuhisa Otani, Shinsaku Tokuda, Mikio Furuse
  • Organizer: 9th Federation of the Asian and Oceanian Physiological Sciences
  • Int'l Joint Research
• [Presentation] Angulin-1 regulates vertical elongation of tricellular tight junction by interacting with scaffolding proteins (2018)
  • Author(s): Taichi Sugawara, Mikio Furuse
  • Organizer: 第70回日本細胞生物学会大会
• [Presentation] ZO family proteins regulate epithelial polarity independent of Tight Junction strand assembly (2018)
  • Author(s): Tetsuhisa Otani, Shinsaku Tokuda, Mikio Furuse
  • Organizer: 第70回日本細胞生物学会大会
• [Remarks] 生理学研究所細胞構造研究部門
  • URL: http://www.nips.ac.jp/dcs/
• [Patent(Industrial Property Rights)] クローディン欠損上皮細胞株の製造方法、クローディン欠損上皮細胞株 (2019)
  • Inventor(s): 古瀬幹夫
  • Industrial Property Rights Holder: 大学共同利用機関法人自然科学研究機構
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2019-226655
  • Filing Date: 2019