April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Actin Reorganization (CLANs) Induced by sCD44-treatment in Trabecular Meshwork Cells
Author Affiliations & Notes
  • Stephen N. Schwartz
    Ophthalmology and Vis Sci, Univ Illinois at Chicago, Chicago, Illinois
  • Rachel M. Beverley
    Ophthalmology and Vis Sci, Univ Illinois at Chicago, Chicago, Illinois
  • Michael C. Giovingo
    Ophthalmology, Cook County Hospital, Chicago, Illinois
  • Ryan D. McCarty
    Ophthalmology and Vis Sci, Univ Illinois at Chicago, Chicago, Illinois
  • Jeffrey P. Mayer
    Ophthalmology and Vis Sci, Univ Illinois at Chicago, Chicago, Illinois
  • Paul A. Knepper
    Ophthalmology and Vis Sci, Univ Illinois at Chicago, Chicago, Illinois
    Dept Ophthalmology, Northwestern University, Chicago, Illinois
  • Footnotes
    Commercial Relationships  Stephen N. Schwartz, None; Rachel M. Beverley, None; Michael C. Giovingo, None; Ryan D. McCarty, None; Jeffrey P. Mayer, None; Paul A. Knepper, Alcon Research Ltd. (R)
  • Footnotes
    Support  Alcon Research Ltd. -- On the Prevention and Treatment of Primary Open-Angle Glaucoma
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 4634. doi:
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    • Get Citation

      Stephen N. Schwartz, Rachel M. Beverley, Michael C. Giovingo, Ryan D. McCarty, Jeffrey P. Mayer, Paul A. Knepper; Actin Reorganization (CLANs) Induced by sCD44-treatment in Trabecular Meshwork Cells. Invest. Ophthalmol. Vis. Sci. 2011;52(14):4634.

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

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Abstract
 
Purpose:
 

Cross-linked reorganization of F-actin networks (CLANs) occurin glaucomatous and steroid-treated trabecular meshwork (TM)cells and may reduce outflow facility. Since soluble CD44 (sCD44)reduces outflow facility in porcine organ perfusion, the purposeof this study was to determine if sCD44 effects CLAN formationin TM cells.

 
Methods:
 

Primary cultures of TM cells were treated with 0.1 ng immunopurifiedsCD44 (n=15) and appropriate controls including heat-inactivatedsCD44 (n=13), stained for F-actin, and visualized with a 100xoil immersion objective on a confocal microscope. The relativeamount of F-actin exhibiting CLAN-like characteristics was quantifiedwithin individual cells. The vertices of the actin (the endsor hubs of F-actin bundles) were identified and counted as exhibitingeither regular formation or CLAN-like formation if at least3 vertices were connected with strongly fluorescent F-actinspokes. The images were partitioned into a grid, and the verticeswere counted in a regular pattern until at least 100 verticeswere counted.

 
Results:
 

An increase in the percent of F-actin arranged in CLANs wasobserved in cells treated with sCD44 (34.75% vertices) as opposedto control cells (11.47% vertices), and the difference was highlysignificant (P<0.00001).  

 

 
Conclusions:
 

Our results indicate that sCD44 induces the formation of CLANsin TM cells. Furthermore, this finding suggests that sCD44 mayalter the outflow resistance of aqueous through its effectson CLAN formation.

 
Keywords: cytoskeleton • glycoconjugates/glycoproteins • trabecular meshwork 
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