Effects of Endothelin-1 on Human Trabecular Meshwork Cell Contraction

Abstract

Trabecular meshwork (TM) cells are now considered to play an active role in the aqueous outflow mechanism, since they exhibit smooth muscle-like contractile properties. Endothelin-1 (ET-1), a potent vasoconstrictor peptide, has been proposed to play a role in the local regulation of aqueous outflow and intra-ocular pressure control. We propose an in vitro culture model as a method in the study of ET-1-induced human trabecular meshwork (HTM) cell contractility. Experiments were performed on semi-confluent HTM cells (primary cultures established from normotensive human donor eyes) at the 2nd passage with PBS as control, ET-1, sarafotoxin 6c (a selective endothelin B receptor agonist) and Y-27632 (a selective inhibitor of rho-associated kinase). The contractile status of the cells was evaluated by a morphometric analysis of the cell area, assuming that HTM cells in culture are able to reduce their area as a consequence of cytoskeletal contraction rather than regulatory volume decrease. Our data indicate that image analysis of the HTM cell area can be a reliable method in the study of TM cell contractility, since it utilizes human cells and offers versatile opportunities for the pharmacological evaluation of drugs controlling HTM cell status.