A new insight into the cellular regulation of aqueous outflow: how trabecular meshwork endothelial cells drive a mechanism that regulates the permeability of Schlemm's canal endothelial cells.
OBJECTIVE
To test the hypothesis that trabecular meshwork endothelial cells (TMEs) increase the permeability of Schlemm's canal endothelial cells (SCEs) by actively releasing ligands that modulate the barrier properties of SCEs.


METHODS
The TMEs were first irradiated with a laser light and allowed to condition the medium, which is then added to SCEs. The treatment response is determined by both measuring SCE permeability (flow meters) and the differential expression of genes (Affymetrix chips and quantitative polymerase chain reaction (PCR)). The cytokines secreted by the treated cells were identified using ELISA and the ability of these cytokines to increase permeability is tested directly after their addition to SCEs in perfusion experiments.


RESULTS
SCEs exposed to medium conditioned by the light activated TMEs (TME-cm) respond by undergoing a differential expression (DE) of 1,120 genes relative to controls. This response is intense relative to a DE of only 12 genes in lasered SCEs. The TME-cm treatment of SCEs increased the SCE permeability fourfold. The role of cytokines in these responses is supported by two findings: adding specific cytokines established to be secreted by lasered TMEs to SCEs increases permeability; and inactivating the TME-cm by boiling or diluting, abrogates these conditioned media permeability effects.


CONCLUSIONS
These experiments show that TMEs can regulate SCE permeability and that it is likely that TMEs have a major role in the regulation of aqueous outflow. This novel TME driven cellular mechanism has important implications for the pathogenesis of glaucoma and the mechanism of action of laser trabeculoplasty. Ligands identified as regulating SCE permeability have potential use for glaucoma therapy.
Alvarado
J A
JA
University of California San Francisco, Department of Ophthalmology; 10 Koret Way; San Francisco, CA 94143-0730, USA. jalva@itsa.ucsf.edu
Alvarado
R G
RG
Yeh
R F
RF
Franse-Carman
L
L
Marcellino
G R
GR
Brownstein
M J
MJ
D001082:Aqueous Humor / Q000502:physiology*; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D016207:Cytokines / Q000378:metabolism / Q000494:pharmacology; D042783:Endothelial Cells / Q000378:metabolism / Q000502:physiology*; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation / Q000528:radiation effects; D006801:Humans; D007834:Lasers; D010539:Permeability / Q000187:drug effects; D012590:Sclera / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D014129:Trabecular Meshwork / Q000166:cytology* / Q000528:radiation effects
D017077:Culture Media, Conditioned; D016207:Cytokines