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2000 Fiscal Year Final Research Report Summary

Physiome and pathophysiome research projects for studying cooperative functions of interstitial space and lymphatic system.

Research Project

Project/Area Number 11470010
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section一般
Research Field General physiology
Research InstitutionShinshu University Graduate School of Medicine

Principal Investigator

OHHASHI Toshio  Shinshu University Graduate School of Medicine, Professor, 大学院・医学研究科, 教授 (80020832)

Co-Investigator(Kenkyū-buntansha) MIZUNO Risuke  Shinshu University School of Medicine, Assistant Professor, 医学部, 助手 (30273080)
IKOMI Fumitaka  Shinshu University School of Medicine, Associate Professor, 医学部, 助教授 (50262704)
Project Period (FY) 1999 – 2000
KeywordsMouse / Intravital Mirosscope / Image Analyzing Appratus / Mesenteric Lymp Circulation / Lymphatic endothelial cell / cell culture / VEGF receptors / NO synthase
Research Abstract

We have designed and constructed a modified intravital microscope system for investigating murine lymphatic pump activity in vivo and developed a custom organ chamber with a semi-circular channel, being suitable for superfusion of murine mesentery in vivo. Using the new experimental appratus, we have evaluated whether or not thore is rhythmic pumping activity of murine mesenteric lymphatic vessels in vivo. A marked lymphatic pumping activity was observed in the mesenteries of DDY mice, The maximal and minimal diameter and frequency in the pumping activity were 60.9±1.0μm, 53.7±1.8μm and 12.8 min, respectively. Physiological roles of endogenous nitric oxide (NO) in the lymphatic pump activity in vivo were studied using the new appratus. A 15-min superfusion of 30μM-NAME in the mesenteries caused significant increasing the pumping frequency and the pump flow index and a significant decrease of the endo-diastoric and end-systolic diameters. Simultaneous superfusion of 1mM L-arginine with … More 30μM L-NAME produced a signficant reversal of the L-NAME-mediated increase of the frequency and decreace of the end-systolic diameter.
We have attempted to examine whether the cultured lymphatic emdothelial cells, similar to angiogenesis of the blood vessels, have an ability to induce in vitro neovascularization of lymph vessels in response to basic fibroblast growth factor (bFGF). The effects of bFGF on the proliferation and migration of the cultured lymphatic endothelial cells isolated from dog thoracic ducts were evaluated by changing the number of the subconfluent cells and by wound migration assay, respectively. The effects of bFGF on invasion and tube formation of the cultured lymphatic endothelial cells into a three-dimensional collagen gel were also investigated by a phase-contrast microscope and an electron microscope. The 10ng/ml bFGF caused a significant induction of proferation and migration of the cultured lymphatic endothelial cells. The bFGF produced significantly invasion and tube formation of the cultured cells into a three-dimensional collagen gel. The bFGF also facilitated to form capillary-like tubes of the cultured cells between twe layers of collagen gets. These findings suggest that the cultured lymphatic endothelial cells have an ability to form lymphatic capillary-like tubes in response to bFGF. Less

  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Nojiri,H.,Ohhashi,T.: "Immunolocalization of nitric oxide synthase and VEGF receptors in cultured lymphatic endothelial cells."Microcirculation. 6. 75-78 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sakai,H.,Ikomi,F.,Ohhashi,T.: "Effects of endothelin on spontaneous contractions in lymph vessels."American Journal of Physiology. 277. H459-H466 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Mizuno,R.,Ono,N.,Ohhashi,T.: "Involvement of ATP-sensitive K^+ channels in spontaneous activity of isolated lymph microvessels in rats."American Journal of Physiology. 277. H1453-H1456 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shirasawa,Y.,Ikomi,F.,Ohhashi,T.: "Physiological roles of endogenous nitric oxide in lymphatic pump activity of rat mesentery in vivo."American Journal of Physiology. 278. G551-G556 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ono,N.,Mizuno,R.,Nojiri,H.,Ohhashi,T.: "Development of experimental apparatus for investigating lymphatic pumping activity of murine mesentery in vivo."Japanese Journal of Physiology. 50. 25-31 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Arai,F.,Mizuno,R.,Ohhashi,T.: "Effects of VEGF on Ca^<2+>-transient in cultured lymphatic endothelial cells and mechanical activity of isolated lymph vessels."Japanese Journal of Physiology. 50. 343-355 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 大橋俊夫(井上正康編): "活性酵素と運動"共立出版株式会社. 301 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 大橋俊夫(日本自律神経学会編): "自律神経機能検査第3版"文光堂. 414 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nojiri, H., Ohhashi, T.: "Immunolocalization of nitric oxide synthase and VEGF receptors in cultured lymphatic endothelial cells."Microcirculation. 6. 75-78 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sakai, H., Ikomi, F., Ohhashi, T.: "Effects of endothelin on spontaneous contractions in lymph vessels."American Journal of Physiology. 277. H459-H466 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Mizuno, R., Ono, N., Ohhashi, T.: "Involvement of ATP-sensitive K^+ channels in spontaneous activity of isolated lymph microvessels in rats."American Journal of Physiology. 277. H1453-H1456 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Shirasawa, Y., Ikomi, F., Ohhashi, T.: "Physiological roles of endogenous nitric oxide in lymphatic pump activity of rat mesentery in vivo."American Journal of Physiology. 278. G551-G556 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ono, N., Mizuno, R., Nojiri, H., Ohhashi, T.: "Development of experimental apparatus for investigating lymphatic pumping activity of murine mesentery in vivo."Japanese Journal of Physiology. 50. 25-31 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Arai, F., Mizuno, R., Ohhashi, T.: "Effects of VEGF on Ca^<2+>-transient in cultured lymphatic endothelial cells and mechanical activity of isolated lymph vessels."Japanese Journal of Physiology. 50. 343-355 (2000)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2002-03-26  

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