Effect of Nerve Growth Factor on Innervation of Perivascular Nerves in Neovasculatures of Mouse Cornea

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  • Matsuyama Akiko
    Department of Clinical Pharmaceutical Science, Graduate School of Medicine Pharmaceutical Sciences, Okayama University
  • Takatori Shingo
    Department of Clinical Pharmaceutical Science, Graduate School of Medicine Pharmaceutical Sciences, Okayama University Department of Clinical Pharmacy, College of Pharmaceutical Sciences, Matsuyama University
  • Sone Yoko
    Department of Clinical Pharmaceutical Science, Graduate School of Medicine Pharmaceutical Sciences, Okayama University
  • Ochi Eiko
    Department of Clinical Pharmaceutical Science, Graduate School of Medicine Pharmaceutical Sciences, Okayama University
  • Goda Mitsuhiro
    Laboratory of Biochemistry, Faculty of Pharmaceutical Sciences, Setsunan University
  • Zamami Yoshito
    Department of Clinical Pharmaceutical Science, Graduate School of Medicine Pharmaceutical Sciences, Okayama University
  • Hashikawa-Hobara Narumi
    Department of Life Science, Okayama University of Science
  • Kitamura Yoshihisa
    Department of Pharmacy, Okayama University Hospital
  • Kawasaki Hiromu
    Department of Clinical Pharmaceutical Science, Graduate School of Medicine Pharmaceutical Sciences, Okayama University Department of Clinical Pharmacy, College of Pharmaceutical Sciences, Matsuyama University

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<p>Angiogenesis, which is the generation of new vascular networks from existing blood vessels, occurs under normal and pathophysiological conditions. Perivascular nerves, which innervate mature vasculatures, maintain vascular tone and regulate tissue blood flow. However, little is known whether perivascular nerves innervate newborn blood vessels. Therefore, the aim of this study was to investigate the distribution and characterization of perivascular nerves in neovasculatures, which were generated by the mouse corneal micropocket method. Under anesthesia, a pellet containing basic fibroblast growth factor (bFGF) (100 ng/pellet) was implanted into a mouse cornea in one side of the eyeball. Nerve growth factor (NGF) was locally (2 or 20 ng) applied with the pellet, or subcutaneously (40 ng/h for 7 d) administered with an osmotic mini-pump. After the implantation, vascular endothelial cells, smooth muscle cells, and perivascular nerves in the cornea were immunohistochemically studied. Neovessels generated from existing limbal vessels were observed in pellet-implanted cornea. Immunostaining of neovasculatures showed the presence of CD31-like immunoreactive (LI) endothelial cells and α-smooth muscle actin-LI vascular smooth muscles. Perivascular nerves immunostained by protein gene product (PGP) 9.5, an axonal marker, were found in the existing limbal vessels, but they were not observed in neovasculatures. Local and subcutaneous treatment of NGF inhibits bFGF-derived angiogenesis and resulted in loop-shaped vessels that had many anastomoses, and produced innervation of PGP 9.5-LI perivascular nerves around bFGF-derived neovessels. These findings suggest that neovasculatures have no innervation of perivascular nerves, and that NGF facilitates innervations of perivascular nerves to regulate the blood flow in neovessels.</p>

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