IOVS Hypertension
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

(Investigative Ophthalmology and Visual Science. 2005;46:375-382.)
© 2005 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.04-0806
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via ISI Web of Science (8)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Walshe, T. E.
Right arrow Articles by Cahill, P. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Walshe, T. E.
Right arrow Articles by Cahill, P. A.

Pulsatile Flow Increases the Expression of eNOS, ET-1, and Prostacyclin in a Novel In Vitro Coculture Model of the Retinal Vasculature

Tony E. Walshe,1 Gail Ferguson,1 Paul Connell,1,2 Colm O’Brien,2 and Paul A. Cahill1

1From the Vascular Health Research Centre, Faculty of Science and Health, Dublin City University, Dublin, Ireland; and the 2Mater Misericordiae Hospital, Institute of Ophthalmology, The Conway Institute of Biomolecular and Biomedical Research, Dublin, Ireland.

PURPOSE. By the development of a novel retinal microvascular endothelial and pericyte cell coculture system, this study determined the effects of pulsatile flow on the activation of the endothelial cell markers nitric oxide (NO), prostacyclin (PGI2), and endothelin (ET)-1.

METHODS. Monocultured bovine retinal endothelial cells (BRECs) and cocultured BRECs with bovine retinal pericytes (BRPs) were exposed to low flow (flow rate, 0.3 mL/min; pulse pressure, 6 mm Hg; shear stress, 0.5 dyne/cm2) or high flow (flow rate, 25 mL/min; pulse pressure, 56 mm Hg; shear stress, 23 dynes/cm2) for 24 hours, by using a novel perfused transcapillary culture system. The cells were characterized by immunohistochemistry and electron and confocal microscopy. Endothelial nitric oxide synthase (eNOS) and phosphorylated-eNOSSer1179 (pp-eNOS) were determined by Western blot analysis. Nitrate, PGI2, and ET-1 levels were quantified in the medium perfusate by using fluorometric and enzyme-linked immunosorbent assays, respectively. Activation of cyclooxygenase (COX)-2 in BRECs was determined by measuring COX-2 promoter activity with a luciferase reporter assay.

RESULTS. The presence of BRPs and BRECs was confirmed by Western blot, immunocytochemistry, and scanning electron microscopy. Phosphorylated eNOS (pp-eNOS) protein levels in BRECs were significantly increased from low to high flow in both mono- and cocultures, concomitant with a significant increase in nitrate levels in the conditioned medium after exposure to pulsatile flow. In parallel cultures, PGI2 levels were also significantly enhanced concomitant with an increase in the transactivation of a COX-2 promoter BREC after exposure to pulsatile flow. ET-1 levels were also increased in both mono- and cocultured cells.

CONCLUSIONS. In this study a novel, functioning, in vitro model of retinal microvascular endothelial and pericyte cells that respond to changes in pulsatile flow was established.




This article has been cited by other articles:


Home page
 J. Appl. Physiol.Home page
A. Uryash, H. Wu, J. Bassuk, P. Kurlansky, M. A. Sackner, and J. A. Adams
Low-amplitude pulses to the circulation through periodic acceleration induces endothelial-dependent vasodilatation
J Appl Physiol, June 1, 2009; 106(6): 1840 - 1847.
[Abstract] [Full Text] [PDF]

Home page
 HypertensionHome page
M. P. Schneider, Y. Ge, D. M. Pollock, J. S. Pollock, and D. E. Kohan
Collecting Duct-Derived Endothelin Regulates Arterial Pressure and Na Excretion via Nitric Oxide
Hypertension, June 1, 2008; 51(6): 1605 - 1610.
[Abstract] [Full Text] [PDF]

Home page
 PhysiologyHome page
B. C. W. Groenendijk, K. Van der Heiden, B. P. Hierck, and R. E. Poelmann
The Role of Shear Stress on ET-1, KLF2, and NOS-3 Expression in the Developing Cardiovascular System of Chicken Embryos in a Venous Ligation Model
Physiology, December 1, 2007; 22(6): 380 - 389.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Renal Physiol.Home page
K. A. Strait, P. K. Stricklett, J. L. Kohan, M. B. Miller, and D. E. Kohan
Calcium regulation of endothelin-1 synthesis in rat inner medullary collecting duct
Am J Physiol Renal Physiol, August 1, 2007; 293(2): F601 - F606.
[Abstract] [Full Text] [PDF]

Home page
 Phil Trans R Soc BHome page
S Moncada
Adventures in vascular biology: a tale of two mediators
Phil Trans R Soc B, May 29, 2006; 361(1469): 735 - 759.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2005 by the Association for Research in Vision and Ophthalmology