Effect of beta-adrenergic agonists on paracellular width and fluid flow across outflow pathway cells.
OBJECTIVE
To determine whether the adrenergic agonists epinephrine and isoproterenol regulate fluid flow across endothelial cells cultured from the human aqueous outflow pathway and to evaluate associated cellular mechanisms.


METHODS
Confluent monolayers of human trabecular meshwork (TM) or Schlemm's canal endothelial (SCE) cells were grown on porous filter supports. The monolayers were perfused with media while fluid flow, expressed as hydraulic conductivity (HC = microl/min/mm Hg/cm2), was continuously measured in preparations treated with isoproterenol, epinephrine, or control medium. Morphometric ultrastructural methods were used to measure the area occupied by the intercellular space and by each cell.


RESULTS
SCE cells and TM cells exposed to isoproterenol or epinephrine responded with an increase in transendothelial fluid flow. Dose-response curves for both adrenergic agonists showed that HC increased linearly as a function of the log of the isoproterenol and epinephrine concentration. At 10(-4) M isoproterenol, the HC increased threefold, and threshold conditions were reached at 10(-9) M. The increase in HC was apparent after isoproterenol had been applied for 1 hour, reached a peak in 3 to 4 hours, and declined gradually to return to baseline conditions in 10 to 12 hours. Morphometric analyses showed that the area occupied by the intercellular space increased fourfold when isoproterenol was used at 10(-4) M, whereas the cell area decreased as a function of the concentration of adrenergic agonist. Epinephrine's effects on HC and cell morphology were blocked by pretreatment with equimolar concentrations of the nonselective beta-blocker, timolol.


CONCLUSIONS
Epinephrine and isoproterenol increase flow through the paracellular pathway of SCE and TM cells through a beta-receptor mediated response that widens the intercellular space and reduces cell area. These findings support the hypothesis that epinephrine decreases the intraocular pressure in glaucoma therapy by promoting fluid flow across the SCE and TM cells lining tissues of the major aqueous outflow pathway.
Alvarado
J A
JA
Department of Ophthalmology, University of California, San Francisco 94143, USA.
Murphy
C G
CG
Franse-Carman
L
L
Chen
J
J
Underwood
J L
JL
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000318:Adrenergic beta-Agonists / Q000494:pharmacology*; D002478:Cells, Cultured; D003470:Culture Media / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004727:Endothelium / Q000166:cytology / Q000187:drug effects* / Q000648:ultrastructure; D004837:Epinephrine / Q000494:pharmacology; D005110:Extracellular Space / Q000187:drug effects*; D006801:Humans; D007365:Intercellular Junctions / Q000187:drug effects*; D007545:Isoproterenol / Q000494:pharmacology; D008855:Microscopy, Electron, Scanning; D010477:Perfusion; D012590:Sclera / Q000166:cytology / Q000187:drug effects* / Q000648:ultrastructure; D014129:Trabecular Meshwork / Q000166:cytology / Q000187:drug effects* / Q000648:ultrastructure
D000318:Adrenergic beta-Agonists; D003470:Culture Media; D007545:Isoproterenol; D015056:1-Methyl-3-isobutylxanthine; D004837:Epinephrine