April 2010
Volume 51, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2010
Differential Expression of Neurotrophins and Nerve Guidance Proteins in Cultured Corneal Fibroblast and Myofibroblasts
Author Affiliations & Notes
  • M. R. Bakir
    Ophthalmology, University of Illinois, Chicago, Chicago, Illinois
  • A. Namavari
    Ophthalmology, University of Illinois, Chicago, Illinois
  • C. Sano
    Ophthalmology, University of Illinois, Chicago, Chicago, Illinois
  • S. Jain
    Ophthalmology, Univ of Illinois Eye and Ear Infirmary, Chicago, Illinois
  • Footnotes
    Commercial Relationships  M.R. Bakir, None; A. Namavari, None; C. Sano, None; S. Jain, None.
  • Footnotes
    Support  EY018874
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 6209. doi:
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      M. R. Bakir, A. Namavari, C. Sano, S. Jain; Differential Expression of Neurotrophins and Nerve Guidance Proteins in Cultured Corneal Fibroblast and Myofibroblasts. Invest. Ophthalmol. Vis. Sci. 2010;51(13):6209.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : We have previously reported expression of nerve guidance cues (Eph/ephrins) on corneal keratocytes. In these experiments our goal is to determine whether the expression of neurotrophins and nerve guidance proteins changes as the keratocyte transitions between wound healing phenotypes (fibroblast & myofibroblast).

Methods: : SV-40 immortalized murine corneal fibroblast cell line was established in 10% FCS culture media and characterized. Fibroblast to myofibroblast transformation was achieved using TGF-beta. Neurotrophins (NGF, GDNF, BDNF, CNTF, NT-3, NT4/5), Nerve guidance proteins (EphB1, ephrinb1), vimentin and alpha-smooth muscle actin levels were assayed by quantitative RT-PCR and quantitative Western analyses (LiCor) in fibroblast and myofibroblast cell lines and compared.

Results: : EphB1/ephrinb1 were differentially expressed. EphB1 expression was decreased in myofibroblasts by 15%, but ephrinb1 was upregulated by 1.7%. Of the neurotrophins BDNF was upregulated in myofibroblasts, but NGF and GDNF were downregulated. Western analysis showed similar trends with ephrinB1 levels increasing by 46% and GDNF decreasing by 25% in myofibroblasts as compared to fibroblasts.

Conclusions: : Neurotrophins and nerve guidance proteins are differentially expressed in different keratocyte phenotypes. Differential regulation of keratocyte derived neurotrophins and nerve guidance proteins may be of significance in the pathogenesis of neurotrophic keratitis.

Keywords: innervation: neural regulation • cornea: stroma and keratocytes • cornea: basic science 
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