Simulated microgravity perturbs actin polymerization to promote nitric oxide-associated migration in human immortalized Eahy926 cells
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Abstract
Microgravity causes endothelium dysfunctions and vascular endothelium remodeling in astronauts returning from space flight. Cardiovascular deconditioning occurs as a consequence of an adaptive response to microgravity partially due to the effects exerted at cellular level. Directional migration of endothelial cell which are central in maintaining the structural integrity of vascular walls is regulated by chemotactic, haptotactic, and mechanotactic stimuli which are essential for vasculogenesis. We explored the migration property of transformed endothelial cells (EC) exposed to 2-h microgravity, simulated using a three-dimensional clinostat constructed based on blueprint published by the Fokker Space, Netherlands. Migration of EC was measured using the scrap wound healing in the presence or absence of actin polymerization inhibitor—cytochalasin D (CD) in Eahy926 cell lines. Simulated microgravity increased cellular migration by 25% while CD-blocked microgravity induced cellular migration. The key migratory structures of cells, filopodia and lamellipodia, formed by EC were more in simulated microgravity compared to gravity. Parallel experiments with phalloidin and diaminorhodamine-4M (DAR-4M) showed that simulated microgravity caused actin rearrangements that lead to 25% increase in nitric oxide production. Further nitric oxide measurements showed a higher nitric oxide production which was not abrogated by phosphoinositol 3 kinase inhibitor (Wortmanin). Bradykinin, an inducer of nitric oxide, prompted two folds higher nitric oxide production along with simulated microgravity in a synergistic manner. We suggest that limited exposure to simulated microgravity increases Eahy926 cell migration by modulating actin and releasing nitric oxide.
Keywords
Simulated microgravity Endothelial cells Actin Nitric oxide MigrationNotes
Acknowledgment
We are grateful to Mr. K.P. Tamilarasan and Mr. Karthikeyan Pasupathy for their technical assistance in fabricating the microgravity machine. We also acknowledge the K.B. Chandrashekar Research Foundation for the financial support.
Conflict of interest
The authors declare that they have no conflict of interest.
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