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Investigative Ophthalmology & Visual Science, Vol 36, 1625-1632, Copyright © 1995 by Association for Research in Vision and Ophthalmology

ARTICLES AND REPORTS

Extravascular secretion of t-PA by the intact superfused choroid

Y Wang, C Gillies, RE Cone and J O'Rourke
Department of Pathology, University of Connecticut Health Center, Farmington 06030-3105, USA.

PURPOSE. Demonstrate the continuous extravascular secretion of enzymatically active t-PA by the intact choroid. METHODS. Enucleated rat eyes were dissected under microscopic control to prepare eyecups for superfusion in a small organ culture system. Individual preparations provided access to intact retinal, retinal pigment epithelial, or choroidal surfaces in situ. Cups were superfused with oxygenated Kreb's buffer (100 microliters/minute) at 37 degrees C and pH 7.4 while suspended in a microincubator. Released t-PA activity was measured by an amidolytic assay in 1-minute overflow samples and in cup media after flow interruptions. RESULTS. Minimal basal secretion was detected in the overflow with the retina in place. Interruption of flow (10 minutes) produced a static accumulation of 0.10 +/- 0.01 IU/ml. With the retinal pigment epithelium in place after retinal removal, t- PA release was moderately accelerated. Removal of the retinal pigment epithelium to expose the Bruch's-choroidal surface produced the greatest acceleration. The 10-minute static level was 0.48 +/- 0.13 IU/ml. Release from the scleral shell after choroid removal was negligible. An acute burst release of t-PA activity (1.3 +/- 0.18 IU/ml) followed the infusion of bradykinin (5 to 20 microM) into choroidal cups. CONCLUSION. Choroid appears to be the dominant source of a continuously secreted t-PA in superfused rat eyecup organ cultures. This enzymatically active t-PA is able to pass through an apparently intact Bruch's membrane but is largely impeded by an intact retinal pigment epithelium. It is also possible that retinal pigment epithelium contains inhibitors that could reduce the released t-PA activity. The authors propose that this secretory function of the uveal tissue, which has thus far not been recognized, may be a principal mechanism for the regulation of intraocular fluid circulation and matrix remodeling by plasmin in vivo.


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J.-M. Rakic, V. Lambert, C. Munaut, K. Bajou, K. Peyrollier, M.-L. Alvarez-Gonzalez, P. Carmeliet, J.-M. Foidart, and A. Noel
Mice without uPA, tPA, or Plasminogen Genes Are Resistant to Experimental Choroidal Neovascularization
Invest. Ophthalmol. Vis. Sci., April 1, 2003; 44(4): 1732 - 1739.
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