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Induction of NEDD4 under simulated microgravity
Author(s) -
Fiedorowicz Katarzyna,
Azevedo Luis Da Silva,
Zakrzewicz Andreas,
Pries Axel Radlach
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.752.8
Subject(s) - shear stress , chemistry , downregulation and upregulation , nedd4 , ubiquitin ligase , biophysics , ubiquitin , microbiology and biotechnology , blot , endothelial stem cell , materials science , biology , biochemistry , gene , in vitro , composite material
Vascular endothelial cells are exposed to mechanical forces e.g. wall shear stress, wall tension, and pulsatile stretch, which are modulated by the gravitational field. The effects of microgravity may influence endothelial cell function including protein synthesis and degradation.Therefore, this study was designed to investigate microgravity dependent effects on the ubiquitin‐ligase NEDD4 in endothelial cells. In order to produce microgravity endothelial cells (EA hy926, HUVEC) were seeded on microcarriers and cultured in the Rotating Wall Vessel for 24 h and 48 h. Controls were kept under normal gravity or exposed to shear stress using a cone and plate apparatus (6 dyn\cm2, 24 h). Gene expression and protein concentration were analysed by real‐time RT‐PCR and Western blotting, respectively. Our results indicate that NEDD4 is upregulated to 200% under microgravity and downregulated to 70% by shear stress relative to controls (1G, no shear). The present data suggest that NEDD4 is sensitive to mechanical forces; the induction under microgravity may be a mechanism to enhance protein degradation in endothelial cells.