Effect of H-7 on cultured human trabecular meshwork cells

[摘要] Purpose: To determine the effect of the serine-threonine kinaseinhibitor H-7, which blocks actomyosin contractility and increasesoutflow facility in live monkeys, on morphology, cytoskeleton, andcellular adhesions of human trabecular meshwork (HTM) cells in culture.Methods: Cultured HTM cells were videographically recorded andevaluated before and after exposure to H-7 at different concentrations.The subcellular distribution of the actin-based cytoskeleton andassociated anchor proteins including vinculin, paxillin, andb-catenin, as well as phosphotyrosine-containing proteins wereevaluated by fluorescence immunocytochemistry and digital fluorescencemicroscopy.Results: H-7 induced pronounced but reversible HTM cell thickeningtoward the cell center and deterioration of the actin cytoskeletalnetwork. Cell-extracellular matrix (ECM) and cell-cell adhesions werealso affected, but the b-catenin-rich, vinculin-containing adherensjunctions were clearly more resistant than focal contacts.Phosphotyrosine labeling in focal contacts was highly sensitive to H-7.Conclusions: H-7 induces alterations in cell shape, actincytoskeleton, and associated focal adhesions in cultured HTM cells,which may be responsible for the effects of H-7 on outflow facility inlive monkey eyes.

[学科分类] 生物化学/生物物理