December 2002
Volume 43, Issue 13
Free
ARVO Annual Meeting Abstract  |   December 2002
Expression of Neurotensin Receptors in Human Corneal Keratocytes
Author Affiliations & Notes
  • T Bourcier
    Ophthalmology 5
    Quinze-Vingts National Center of Ophthalmology Paris France
  • N Rondeau
    Ophthalmology 5
    Quinze-Vingts National Center of Ophthalmology Paris France
  • S Paquet
    U339 Saint-Antoine Hospital Paris France
  • P Forgez
    U339 Saint-Antoine Hospital Paris France
  • A Lombet
    U339 Saint-Antoine Hospital Paris France
  • F Pouzaud
    Cellular pharmacotoxicology unit
    Quinze-Vingts National Center of Ophthalmology Paris France
  • W Rostène
    U339 Saint-Antoine Hospital Paris France
  • V Borderie
    Ophthalmology 5
    Quinze-Vingts National Center of Ophthalmology Paris France
  • L Laroche
    Ophthalmology 5
    Quinze-Vingts National Center of Ophthalmology Paris France
  • Footnotes
    Commercial Relationships   T. Bourcier, None; N. Rondeau, None; S. Paquet, None; P. Forgez, None; A. Lombet, None; F. Pouzaud, None; W. Rostène, None; V. Borderie, None; L. Laroche, None.
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 3209. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      T Bourcier, N Rondeau, S Paquet, P Forgez, A Lombet, F Pouzaud, W Rostène, V Borderie, L Laroche; Expression of Neurotensin Receptors in Human Corneal Keratocytes . Invest. Ophthalmol. Vis. Sci. 2002;43(13):3209.

      Download citation file:


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

      ×
  • Supplements
Abstract

Abstract: : Purpose:The aim of the study was to investigate whether cultured human keratocytes express the neurotensin receptors (NTR1, NTR2, and NTR3), to determine the presence of neurotensin (NT) in keratocytes, and to assess the influence of NT on these cells. Methods:Human keratocytes were cultured in culture medium treated with various concentrations (10-7 to 10-9 M) of JMV449 (a weakly degradable NT agonist). Cell proliferation and viability were analyzed by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt (MTS) assay. Apoptosis was studied by nucleus labeling using a fluorescent dye (Hoescht 33342) and cold light fluorometry. NT, NTR1, NTR2, NTR3 mRNA were detected in human keratocytes by means of reverse transcriptase polymerase chain reaction (RT-PCR). NTR1 protein was detected by Western-blot analysis. Functionality of NTR1 was assessed by intracellular calcium measurement using a dynamic imaging microscopy system. Results:RT-PCR and Western-blot showed the expression of the NTR1 (mRNA and protein), and NTR3 mRNA in human corneal keratocytes. NT and NTR2 mRNA were undetectable. JMV449 induced a rapid and transient intracellular calcium increase in human corneal keratocytes that was blocked by the specific antagonist SR48692. JMV449 significantly increased cell proliferation and viability after 72, 96 and 120 hours of culture with a maximum effect at 10-7 M (p < 0.005). Finally, JMV449 decreased keratocyte apoptosis whatever the concentration used (p < 0.005). Conclusion:These results indicate that cultured human keratocytes express NTR1 and NTR3, and that NT may exert physiological effects on cornea such as regulation of keratocyte proliferation and apoptosis.

Keywords: 488 neuropeptides • 374 cornea: stroma and keratocytes • 370 cornea: basic science 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×