May 2003
Volume 44, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2003
Growth Factors in Secondary Aqueous Humor Modulate Trabecular Cell Expression of MMP-3
Author Affiliations & Notes
  • R.C. Tripathi
    Ophthalmology, USC-SOM Vision Research Lab, Columbia, SC, United States
  • B.J. Tripathi
    Pathology & Microbiology, USC-SOM, Columbia, SC, United States
  • J. Chen
    Pathology & Microbiology, USC-SOM, Columbia, SC, United States
  • J. Li
    Ophthalmology, USC-SOM, Columbia, SC, United States
  • K.V. Chalam
    Ophthalmology, University of Florida, Jacksonville, FL, United States
  • S. Gotsis
    Ophthalmology, University of Athens, Athens, Greece
  • V.A. Pakalnis
    Ophthalmology, University of Athens, Athens, Greece
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 1167. doi:
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      R.C. Tripathi, B.J. Tripathi, J. Chen, J. Li, K.V. Chalam, S. Gotsis, V.A. Pakalnis; Growth Factors in Secondary Aqueous Humor Modulate Trabecular Cell Expression of MMP-3 . Invest. Ophthalmol. Vis. Sci. 2003;44(13):1167.

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Abstract

Abstract: : Purpose: To determine the mRNA and protein expression of matrix metalloprotease (MMP)-3 (stromelysin-1) by trabecular cells treated with growth factors present in secondary aqueous humor encountered in glaucomatous disease of the eye. Methods: Serum-starved, first passage porcine trabecular cells were incubated from 48 hours to 7 days in medium containing the combination and concentration of growth factors that are present in secondary aqueous humor and, as controls, in serum-free medium. We extracted total RNA, performed RT-PCR using primer pairs specific for porcine MMP-3 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and quantified the products. We utilized Western blotting to detect and quantify MMP-3 protein. Results: Compared to controls, expression of MMP-3 mRNA by trabecular cells was down-regulated 70% after incubation for 48 hours as well as 7 days in medium containing growth factors present in secondary aqueous humor. MMP-3 protein was not detectable by western blotting. Conclusions: Matrix metalloproteases have a crucial role in the regulated turnover of the extracellular matrix (ECM) in the trabecular meshwork. The down-regulation of MMP-3 mRNA by trabecular cells exposed to growth factors present in secondary aqueous humor implies a decreased proteolytic activity that contributes to accumulation of components of the ECM, such as glycoproteins and proteoglycans. Our findings indicate that induction of MMP-3 gene expression in the trabecular meshwork of glaucomatous eyes could effectively reduce buildup of ECM molecules in the aqueous outflow pathway to increase outflow facility.

Keywords: growth factors/growth factor receptors • extracellular matrix • outflow: trabecular meshwork 
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