Analysis of the mechanism that controls vertebrates'' inner ear/semicircular canal morphogenesis

• Project/Area Number: 18570192
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Developmental biology
• Research Institution: Tohoku University
• Principal Investigator: FUNAHASHI Jun-ichi Tohoku University, Institute of Development, Aging & Cancer, Dept. of Mol Neurobiol, Associate Proffessor (00270827)
• Project Period (FY): 2006 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥4,110,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥510,000); Fiscal Year 2007: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2006: ¥1,900,000 (Direct Cost: ¥1,900,000)
• Keywords: zebrafish / gallery / mutant / transgeneic / 3D time-lapse / positional cloning / splicing valiant / 内耳 / 三半規管 / ニワトリ / BMP / Nogin / GFP

Research Abstract

Vertebrate's semicircular canals in inner ear are intriguing model systems to study the mechanism of morphogenesis, Because their function and conserved among vertebrates.
In the semicircular canals of chick embryo, we made a precise immunohistochemical analysis, and got results suggesting that epithelial-mesenchymal transition plays a roll though it had been thought to be the least plausible mechanism.
By screening zebrafish mutants, we have isolated a mutant line named gallery that has disorganized semicircular canal. In the wild-type, the formation of semicircular canals begins with the development of protrusions that extend inward form the wall of otic vesicle into the lumen. While in gallery, only one protrusion is formed. We are mapping the responsible gene in genome and will be able to clone the gene in coning year.
To study semicircular canal morphogenesis with the resolution of cellular level, we have attempted isolate the fish that specifically express GFP in the otic vesicle. By screening Gal4 gene trap lines, we have identified the line that expresses GFP at the tip of the growing protrusions. We made three-dimensional time-lapse analysis, along with the identification of Gal4 insertion point and cloning the neighboring gene. The cloned gene is a know gene, although it consist of upstream exons. This splicing form might have different expression pattern form conventional splicing form, and we now comparing their expression patterns. We are planning to knock down this specific splicing form and study its function in inner ear morphogenesis.

Research Products

• [Journal Article] Epithelial-mesenchymal transition as a possible mechanism of semicircular canal morphogenesis in chick inner ear (2008)
  • Author(s): Kobayashi, Y., Nakamura, H.&Funahashi, J.
  • Journal Title: The Tohoku Journal of Experimental Medicine (印刷中)
  • NAID: 10024168349
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Dan is required for normal morphogenesis and patterning in the developing chick inner ear. (2007)
  • Author(s): Takahiro Yamanishi, Kenjiro Katsu, Jun-ichi Funahashi, Eiji Yumoto and Yuji Yokouchi
  • Journal Title: Develop. Growth Differ. 49 Pages: 13-26
  • NAID: 10018521158
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Role of Bone morphogenetic protein 4 in zebrafish semicircular canal development. (2007)
  • Author(s): Omata, Y., Nojima, Y., Nakayama, S., Okamoto, H., Nakamura, H.&Funahashi, J.
  • Journal Title: Dev. Growth Differ 49 Pages: 711-719
  • NAID: 10019984157
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Role of Bone morphogenetic protein 4 in zebrafish semicircular canal development (2007)
  • Author(s): Omata, Y., Nojima, Y., Nakayama, S., Okamoto, H., Nakamura, H. & Funahashi, J.
  • Journal Title: Dev. Growth Differ 49 Pages: 711-719
  • NAID: 10019984157
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Dan is required for normal morphogenesis and patterning in the developing chick inner ear (2007)
  • Author(s): Takahiro Yamanishi, Kenjiro Katsu, Jun-ichi Funahashi, Eiji Yumoto and Yuji Yokouchi
  • Journal Title: Dev. Growth Differ 49 Pages: 13-26
  • NAID: 10018521158
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Role of Bone morphogenetic protein 4 in zebrafish semicircular canal development. (2007)
  • Author(s): Omata, Y., Nojima, Y., Nakayama, S., Okamoto, H., Nakamura, H. & Funahashi, J.
  • Journal Title: Dev. Growth Differ 49 Pages: 711-719
  • NAID: 10019984157
  • Peer Reviewed
• [Journal Article] Dan is required for normal morphogenesis and patterning in the developing chick inner ear. (2007)
  • Author(s): Takahiro Yamanishi, Kenjiro Katsu, Jun-ichi Funahashi, Eiji Yumoto, Yuji Yokouchi
  • Journal Title: Develop. Growth Differ. 49 Pages: 13-26
  • NAID: 10018521158
• [Journal Article] Role of Gbx2 and Otx2 in the formation of cochlear ganglionハand endolymphatic duct (2006)
  • Author(s): Hiromitsu Miyazaki, Toshimitsu Kobayashi, Harukazu Nakamura and Jun-ichi Funahashi
  • Journal Title: Develop. Growth Differ. 48 Pages: 429-438
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Role of Gbx2 and Otx2 in the formation of cochlear ganglion and endolymphatic duct (2006)
  • Author(s): Hiromitsu Miyazaki, Toshimitsu Kobayashi, Hamkazu Nakamura and Jun-ichi Funahashi
  • Journal Title: Dev. Growth Differ 48 Pages: 429-438
  • NAID: 10020141479
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Role of Gbx2 and Otx2 in the formation of cochlear ganglion and endolymphatic duct (2006)
  • Author(s): Hiromitsu Miyazaki, Toshimitsu Kobayashi, Harukazu Nakamura, Jun-ichi Funahashi
  • Journal Title: Develop. Growth Differ. 48 Pages: 429-438
  • NAID: 10020141479
• [Journal Article] Epithelial-mesenchymal transition as a possible mechanism of semicircular canal morphogenesis in chick inner ear
  • Author(s): Kobayashi, Y., Nakamura, H. & Funahashi. J
  • Journal Title: Tohoku J. Exp. Med.
  • NAID: 10024168349
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 遺伝子トラップ系統を用いたゼブラフィッシュ三半規管形態形成の解析 (2007)
  • Author(s): 舟橋淳一、川上浩一、仲村春和
  • Organizer: 第30回日本分子生物学会年会
  • Place of Presentation: 横浜(パシフィコ横浜)
  • Year and Date: 2007-12-14
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Analysis of zebrafish semicircular canal morphogenesis by gene trap line (2007)
  • Author(s): Funahashi, J., Kawakamai, K. & Nakamura, H
  • Organizer: 30th Annual Meeting of the Molecular Biology Society of Japan
  • Place of Presentation: Yokohama
  • Year and Date: 2007-12-14
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 三次元タイムラプスによるゼブラフィッシュ三半規管形態形成の解析 (2007)
  • Author(s): 舟橋淳一、川上浩一、仲村春和
  • Organizer: 第13回小型魚類研究会
  • Place of Presentation: 東京大学(小柴ホール)
  • Year and Date: 2007-09-17
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 3D time-lapse analysis of zebrafish semicircular canal morphogenesis (2007)
  • Author(s): Funahashi, J., Kawakamai, K. & Nakamura, H
  • Organizer: 13th Medaka and Zebrafish Meeting
  • Place of Presentation: Tokyo
  • Year and Date: 2007-09-17
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 遺伝子トラップ系統を用いたゼブラフィッシュ三半規管形態形成の解析 (2007)
  • Author(s): 舟橋 淳一, 川上 浩一, 仲村 春和
  • Organizer: 第40回日本発生生物学会
  • Place of Presentation: 福岡(国際会議場)
  • Year and Date: 2007-05-29
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Analysis of zebrafish semicircular canal morphogeneisi by gene trap line (2007)
  • Author(s): Funahashi, J., Kawakamai, K. & Nakamura, H.
  • Organizer: 40th Annual Meeting of the Japanese society of Developmental Biologisits
  • Place of Presentation: Fukuoka
  • Year and Date: 2007-05-29
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 三半規管形成に異常を示すゼブラフィッシュ突然変異体galleryとBMPシグナルについて (2006)
  • Author(s): 小又 尉広, 野島 康弘, 中山 里実, 岡本 仁, 仲村 春和, 舟橋 淳一
  • Organizer: 第39回日本発生生物学会
  • Place of Presentation: 広島(国際会議場)
  • Year and Date: 2006-06-01
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 三半規管発生の組織学的検討 (2006)
  • Author(s): 小林 友理子, 仲村 春和, 舟橋 淳一
  • Organizer: 第39回日本発生生物学会
  • Place of Presentation: 広島(国際会議場)
  • Year and Date: 2006-06-01
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] BMP signaling and zebrafish mutant gallery that has semicircular canal defects (2006)
  • Author(s): Omata, Y., Nojima, Y., Nakayama, S., Okamoto, H., Nakamura, H., & Funahashi, J.
  • Organizer: 39th Annual Meeting of the Japanese society of Development Biologisits
  • Place of Presentation: Hiroshima
  • Year and Date: 2006-06-01
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Immunohisochemical analysis of the semicircular canal development (2006)
  • Author(s): Kobayashi, Y., Nakamura, H. & Funahashi, J.
  • Organizer: 39th Annual Meeting of the Japanese society of Developmental Biologisits
  • Place of Presentation: Hiroshima
  • Year and Date: 2006-06-01
  • Description: 「研究成果報告書概要(欧文)」より