The role of actomyosin in the regulation of syndecan-1 in hyperosmosis.

Introduction: The endothelial glycocalyx is susceptible to high concentration of glucose and sodium in the blood. These challenges often involve an increase in osmotic pressure which may independently alters the glycocalyx components. The glycocalyx anchors on the cell membrane via core proteins that link with the actin cytoskeleton. This study aims to investigate the role of actomyosin in the osmoregulation of syndecan-1, a core protein that bears abundant sugar chains of the glycocalyx.

Methods: Human umbilical vein endothelial cells were incubated with mannitol-based hyperosmotic medium up to 2 h. The surface expression of syndecan-1 and the actin cytoskeleton were analysed by confocal microscopy, either without or with cytoskeletal manipulation.

Results: Syndecan-1 expression was compromised when hyperosmotic challenge was prolonged for 2 h, with the normalised intensity substantially dropped to 65.78 ± 2.07% at +200 mOsm. The reduction is associated with a sustained actin hyper-polymerisation, including significant increases in cortex coverage and cytoskeletal tension. Disassembling the cortex by cytochalasin D restores syndecan-1 in hyperosmosis. Inhibition of ROCK, rather than MLCK and myosin II ATPase activity, prevents the reduction of syndecan-1.

Conclusion: We have demonstrated that prolonged hyperosmotic stress disrupts the integrity of syndecan-1 through an aberrant cortex polymerisation. Our results provide new evidence in the interplay between the glycocalyx and the actin. It helps us better interpret the regulation of the glycocalyx, moving towards a goal of protecting and restoring the glycocalyx under healthy and diseased conditions.