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2003 Fiscal Year Final Research Report Summary

Identification of genes involved in emerging cellular properties of corneal epithelial stem cells and its specific differentiation

Research Project

Project/Area Number 14370563
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Ophthalmology
Research InstitutionKyoto Prefectural University of Medicine

Principal Investigator

KINOSHITA Shigeru  Kyoto Prefectural University of Medicine, Medicine, Ophthalmology, Professor, 医学研究科, 教授 (30116024)

Co-Investigator(Kenkyū-buntansha) KAWASAKI Satoshi  Kyoto Prefectural University of Medicine, Medicine, Ophthalmology, Professor, 医学研究科, 助手 (60347458)
Project Period (FY) 2002 – 2003
Keywordscornea / limbus / basal cells / stem cell / iAFLP / gene expression / NGFR (p75NTR) / TrkA
Research Abstract

To identify genes to render special cellular properties in stem cells of corneal epithelium, we performed gene expression analysis among 3 layers of corneal, limbal and conjunctival epithelial cells. For selective collection of limbal basal epithelial cells which are thought to be putative stem cells of corneal epithelial cells, we used a laser micro-dissection device. For such analysis, the major obstacles are limited amount of sample ; the estimated maximum yield of RNA might be as much as 1ng of total RNA. Thus we optimized iAFLP method for precise determination of gene expression profile from tiny amounts of RNA, which were extracted from laser dissected frozen tissues fragments. After several trial and error experiments, we confirmed the optimized iAFLP method to be adjusted to such small RNA. Gene expression analysis among the 9 samples against approx 500 corneal epithelium-specific genes allowed the identification of several genes which were specifically expressed in limbal basal epithelial cells. These genes included integrin alpha6,alpha7,calcium activated chloride channel 2 (CLCA2), NGF receptor (p75NTR), BAD, TNFR2,THBS3 and keratin 18. Among them, p75 NTR exhibited a very intriguing tissue distribution when we observed its expression in cornea and limbus by immunohistochemistry. Tissue distribution of the gene showed dominant and specific expression in limbal basal epithelial cells, suggesting the possibility of this gene as a specific marker for the corneal epithelial stem cells. Furthermore, its counterpart gene, TrkA, exhibited a very consistent distribution to the p75NTR, strongly implying the possibility of the potential function of NOF, a neurotrophic growth factor and a ligand for these dimeric receptors in maintaining and rendering the stem cell property.

  • Research Products

    (11 results)

All Other

All Publications (11 results)

  • [Publications] Nakamura et al.: "Successful regrafting of cultivated corneal epithelium using amniotic membrane as a carrier in severe ocular surface disease."Cornea. 22(1). 70-71 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamada et al.: "Local suppression of IL-1 by receptor antagonist in the rat model of corneal alkali injury"Experimental Eye Research. 76(2). 161-167 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ban et al.: "Tight junction-related protein expression and distribution in human."Experimental Eye Research. 76(6). 663-669 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ban et al.: "Comparison of ultrastructure, tight junction-related protein expression and barrier function of human corneal epithelial cells cultivated on amniotic membrane with and without air-lifting"Experimental Eye Research. 76(6). 735-743 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kawasaki et al.: "Up-regulated gene expression in the conjunctival epithelium of patients with Sjogren's syndrome."Experimental Eye Research. 77(1). 17-26 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakamura et al.: "Successful regrafting of cultivated corneal epithelium using amniotic membrane as a carrier in severe ocular surface disease."Cornea. 22-1. 70-71 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamada et al.: "Local suppression of IL-1 by receptor antagonist in the rat model of corneal alkali injury"Experimental Eye Research. 76-2. 161-167 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ban et al.: "Tight junction-related protein expression and distribution in human."Experimental Eye Research. 76-6. 663-669 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ban et al.: "Comparison of ultrastructure, tight junction-related protein expression and barrier function of human corneal epithelial cells cultivated on amniotic membrane with and without air-lifting."Experimental Eye Research. 76-6. 663-669 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kawasaki et al.: "Up-regulated gene expression in the conjunctival epithelium of patients with Siogren's syndrome."Experimental Eye Research. 77-1. 17-26 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Maruyama et al.: "TH2-biased immune system promotion of allogeneic corneal epithelial cell survival after orthotopic limbal transplantation."Invest Ophthal Vis Sci. 44-11. 4736-4741 (2003)

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

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Published: 2005-04-19  

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