May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Cochlin-Protein Interactions and Its Potential Transcriptional Regulation in Glaucomatous Trabecular Meshwork
Author Affiliations & Notes
  • R. Picciani
    Miller School of Medicine, Bascom Palmer Eye Institute, Miami, Florida
  • S. K. Bhattacharya
    Miller School of Medicine, Bascom Palmer Eye Institute, Miami, Florida
  • Footnotes
    Commercial Relationships  R. Picciani, None; S.K. Bhattacharya, None.
  • Footnotes
    Support  EY15266, EY16112, P30EY14801, an unrestricted center grant from Research to Prevent Blindness (RPB) to University of Miami and RPB Career Award (SKB).
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 1644. doi:https://doi.org/
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      R. Picciani, S. K. Bhattacharya; Cochlin-Protein Interactions and Its Potential Transcriptional Regulation in Glaucomatous Trabecular Meshwork. Invest. Ophthalmol. Vis. Sci. 2008;49(13):1644. doi: https://doi.org/.

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

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Abstract

Purpose: : To identify the cochlin interacting proteins in the glaucomatous trabecular meshwork (TM) and to demonstrate potential differences in the transcriptional regulation machinery for cochlin expression in the glaucomatous TM compared to controls.

Methods: : Normal human and POAG donor eyes (n=20 for each group) were obtained from NDRI in accordance with the declaration of Helsinki. Dissected TM tissues were subjected to protein extraction, Western analysis and overlay assays. Electrophoretic mobility shift assays (EMSA) were performed using biotin labeled oligonucleotides (bearing transcription factor binding sites in the cochlin promoter region identified by bioinformatics) and TM nuclear extracts (glaucomatous and control).

Results: : Annexin A2 was identified as cochlin interacting protein in the glaucomatous TM by mass spectrometry and confirmed by reciprocal immunoprecipitation. Expression of cochlin in HEK cells, but not annexin, led to secretion of annexin. Glaucomatous TM in contrast to normal TM showed elevated levels of transcription factors including Brn-3 by EMSA and Western analysis.

Conclusions: : Annexin A2 interacts with cochlin in glaucomatous TM. Protein expression studies (in mammalian cells) and elevated levels of select transcription factors in glaucomatous TM compared to control further supports transcriptional control of cochlin-interacting proteins in the glaucomatous TM.

Keywords: trabecular meshwork • extracellular matrix • transcription factors 
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