June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
PEDF Effects on Outflow Facility
Author Affiliations & Notes
  • Morgan Rogers
    Duke University, Durham, NC
  • Iris Navarro
    Duke University, Durham, NC
  • Kristin Perkumas
    Duke University, Durham, NC
  • R Rand Allingham
    Duke University, Durham, NC
  • Pratap Challa
    Duke University, Durham, NC
  • Craig Crosson
    Medical University of South Carolina, Charleston, SC
  • W Daniel Stamer
    Duke University, Durham, NC
  • Footnotes
    Commercial Relationships Morgan Rogers, None; Iris Navarro, None; Kristin Perkumas, None; R Rand Allingham, New World Medical (C); Pratap Challa, None; Craig Crosson, Alimera Sciences (C), Lexicon Pharmaceuticals, Inc (R); W Daniel Stamer, Allergan (F), Alcon (F), Acucela (C), Aerie (C), Cytokinetics (C)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 2004. doi:
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    • Get Citation

      Morgan Rogers, Iris Navarro, Kristin Perkumas, R Rand Allingham, Pratap Challa, Craig Crosson, W Daniel Stamer; PEDF Effects on Outflow Facility. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2004.

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

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Abstract

Purpose: Preliminary data from our laboratory show that pigment epithelium derived factor (PEDF) increases transendothelial electrical resistance of human SC and porcine AAP endothelial monolayers in a time and dose-dependent manner. In this study, our goal was to determine PEDF levels in human aqueous humor, and the effect of PEDF on outflow function in enucleated mouse eyes.

Methods: Aqueous humor was obtained from consented patients during ocular surgery. Fresh samples were immediately placed on dry ice and kept frozen at -80C until an ELISA PEDF assay (Chemikine) was performed. Eyes from culled C57BL/6 mice were enucleated and cannulated for ex vivo perfusion using a computer-controlled perfusion system optimized for mouse eyes. Purified PEDF (1μg/ml) was perfused at four different pressures (4, 8, 15, 25 mmHg), measuring flow to determine outflow facility (slope of flow/pressure relationship). Data were compared to eyes perfused with vehicle alone (negative control) and two positive controls to establish range for detection of changes in outflow facility: dithia PGE-1 (10 nM) and sphingosine-1-phosphate (S1P, 5μM).

Results: Eighteen human aqueous humor samples were examined and found to contain PEDF at levels ranging from 0.12-10.72 μg/mL (1.519 ± 2.492). We did not detect a relationship between PEDF level and gender, race or donor glaucoma status. We perfused a total of 27 C57BL/6 mouse eyes. Compared to vehicle-perfused controls, we observed an 86% increase in outflow facility for dithia PGE-1 (0.035 vs. 0.019 μl/min/mmHg), and a 20% decrease in outflow facility for S1P (0.017 vs. 0.022 μl/min/mmHg), similar to values previously reported. We perfused 7 eyes with PEDF and observed an average outflow facility of 0.018 μl/min/mmHg compared to 0.019 μl/min/mmHg for controls. Interestingly, there appeared to be two sets of responses within our data: 4 eyes having mean outflow facility of 0.010 ± 0.005 μl/min/mmHg, and the other 3 with mean of 0.028 ± 0.001 μl/min/mmHg.

Conclusions: PEDF is present in human aqueous humor at physiologically relevant concentrations. On average PEDF did not significantly affect outflow facility. However, the data set was bimodal with 4 eyes decreasing outflow facility by 50%, and the other 3 increasing outflow facility by 67%. We do not presently understand this phenomenon but have only tested one concentration of PEDF which was effective in vitro but may be too high ex vivo. We plan to perform a full concentration-response.

Keywords: 427 aqueous • 735 trabecular meshwork • 543 growth factors/growth factor receptors  
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