In vivo calcium imaging analyses during angiogenesis and vessel stabilization

• Project/Area Number: 15K18976
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: General physiology
• Research Institution: National Cardiovascular Center Research Institute
• Principal Investigator: Nakajima Hiroyuki 国立研究開発法人国立循環器病研究センター, 研究所, 室長 (10467657)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2016: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 細胞内カルシウム / 血管新生 / メカニカルストレス / 血管形成 / 血流 / カルシウム / 血管内皮増殖因子

Outline of Final Research Achievements

In this study, we visualized endothelial Ca2+ dynamics at single-cell resolution in zebrafish and found that endothelial Ca2+ oscillations completely depended upon VEGF-A/VEGFR2 signaling during intersomitic vessel (ISV) formation. Thus, visualizing Ca2+ oscillations allowed us to monitor indirectly how VEGF-A/VEGFR2 signaling is spatio-temporally regulated at single cell level. We also observed blood flow-dependent Ca2+ oscillations in perfused vessels. By analyzing endothelial responses to blood flow, we newly found that YAP, a　final effector of the Hippo pathway, responds to flow-mediated mechanical stimuli.

Research Products

• [Journal Article] Flow-dependent endothelial YAP regulation that contributes to vessel maintenance. (2017)
  • Author(s): Nakajima H, Yamamoto K, Agarwala S, Terai K, Fukui H, Fukuhara S, Ando K, Miyazali T, Yokota Y, Sxhmelzer E, Belting H, Affolter M, Lecaudey, Mochizuki N
  • Journal Title: Developmental Cell Volume: 40 Issue: 6 Pages: 523-536
  • DOI: 10.1016/j.devcel.2017.02.019
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Polydom is an extracellular matrix protein involved in lymphatic vessel remodeling. (2017)
  • Author(s): Morooka, N., Futaki, S., Sato-Nishiuchi, R., Nishino, M., Totani, Y., Shimono, C., Nakano, I., Nakajima, H., Mochizuki, N., and Sekiguchi, K.
  • Journal Title: Circ. Res. Volume: 120 Issue: 8 Pages: 1276-1288
  • DOI: 10.1161/circresaha.116.308825
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] In vivo visualization of cardiomyocyte apicobasal polarity reveals epithelial to mesenchymal-like transition during cardiac trabeculation. (2016)
  • Author(s): Vanesa Jimenez-Amilburu, S. Javad Rasouli, David W. Staudt, Hiroyuki Nakajima, Ayano Chiba, Naoki Mochizuki, Didier Y.R. Stainier
  • Journal Title: Cell Repot Volume: 17 Issue: 10 Pages: 2687-2699
  • DOI: 10.1016/j.celrep.2016.11.023
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Clarification of mural cell coverage of vascular endothelial cells by live imaging of zebrafish. (2016)
  • Author(s): Ando K, Fukuhara S, Izumi N, Nakajima H, Fukui H, Kelsh RN, Mochizuki N.
  • Journal Title: Development Volume: 143 Pages: 1328-1339
  • DOI: 10.1242/dev.132654
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Endothelial Ca 2+ oscillations reflect VEGFR signaling-regulated angiogenic capacity in vivo (2015)
  • Author(s): Yokota Y*, Nakajima H*(Co-first author), Wakayama Y, Muto A, Kawakami K, Fukuhara S, Mochizuki N.
  • Journal Title: eLife Volume: 4
  • DOI: 10.7554/elife.08817
  • Peer Reviewed / Open Access
• [Journal Article] Recent advances in understanding of cardiovascular development by imaging of zebrafish. (2015)
  • Author(s): Fukuhara S, Fukui H, Wakayama Y, Ando K, Nakajima H, Mochizuki N.
  • Journal Title: Dev Growth Differ. Volume: 未定 Issue: 4 Pages: 109-122
  • DOI: 10.1111/dgd.12210
  • Peer Reviewed / Open Access
• [Presentation] Endothelial YAP is regulated by flow-mediated mechanical stimuli through the regulation of F-actin and contributes to vessel maintenance （口頭発表） (2016)
  • Author(s): 〇中嶋洋行、望月直樹
  • Organizer: 第24回日本血管生物医学会学術集会
  • Place of Presentation: 長崎ブリックホール・長崎
  • Year and Date: 2016-12-08
• [Presentation] 血管新生過程における内皮細胞応答のin vivoイメージング解析/ In vivo imaging analyses of endothelial responses to signaling inputs during angiogenesis （若手招聘講演） (2016)
  • Author(s): 〇中嶋洋行、横田泰宏、望月直樹
  • Organizer: 第15回日本心臓血管発生研究会
  • Place of Presentation: 国立循環器病研究センター・大阪
  • Year and Date: 2016-10-14
  • Invited
• [Presentation] Visualization of endothelial VEGFR2-dependent Ca2+ dynamics during sprouting angiogenesis （ポスター発表) (2016)
  • Author(s): 〇中嶋洋行、横田泰宏、福原茂朋、望月直樹
  • Organizer: 9th International Kloster Seeon Meeting
  • Place of Presentation: Kloster Seeon, Germany
  • Year and Date: 2016-09-17
  • Int'l Joint Research
• [Presentation] VEGFR2 活性化の間接的可視化による血管新生メカニズムの解明/Analysis of sprouting angiogenesis by indirectly visualizing VEGFR2 activity in vivo （口頭発表） (2016)
  • Author(s): 〇中嶋洋行、横田泰宏、福原茂朋、望月直樹
  • Organizer: 第68回日本細胞生物学会
  • Place of Presentation: 京都テルサ・京都
  • Year and Date: 2016-06-15
• [Presentation] VEGFR2 活性化の間接的可視化による血管新生メカニズムの解明【口頭発表】 (2016)
  • Author(s): 中嶋洋行、横田泰宏、福原茂朋、望月直樹
  • Organizer: 第68回日本細胞生物学会
  • Place of Presentation: 京都
  • Year and Date: 2016-06-15
• [Presentation] Flow-dependent mechanical stress induces nuclear translocation of YAP1 by regulating actin cytoskeleton【ポスター発表】 (2015)
  • Author(s): Hiroyuki Nakajima, Kimiko Yamamoto, Hajime Fukui, Shigetomo Fukuhara and
  • Organizer: Gordon Research Conference
  • Place of Presentation: Newport
  • Year and Date: 2015-08-02
  • Int'l Joint Research
• [Presentation] Flow-dependent mechanical stress induces nuclear translocation of YAP1 by regulating actin cytoskeleton【口頭発表】 (2015)
  • Author(s): Hiroyuki Nakajima, Kimiko Yamamoto, Hajime Fukui, Shigetomo Fukuhara and
  • Organizer: Gordon Research Seminer
  • Place of Presentation: Newport
  • Year and Date: 2015-08-01
  • Int'l Joint Research
• [Presentation] 血流による機械的刺激に対するYAP1の応答機構【口頭発表】 (2015)
  • Author(s): 中嶋洋行、山本希美子、福井一、福原茂朋、望月直樹
  • Organizer: 第67回日本細胞生物学会
  • Place of Presentation: 東京
  • Year and Date: 2015-06-30