cDNA microarray reveals the alterations of cytoskeleton-related genes in osteoblast under high magneto-gravitational environment

Abstract

The diamagnetic levitation as a novel ground-based model for simulating a reduced gravity environment has been widely applied in many fields. In this study, a special designed superconducting magnet, which can produce three apparent gravity levels (0, 1, and 2 g), namely high magneto-gravitational environment (HMGE), was used to simulate space gravity environment. The effects of HMGE on osteoblast gene expression profile were investigated by microarray. Genes sensitive to diamagnetic levitation environment (0 g), gravity changes, and high magnetic field changes were sorted on the basis of typical cell functions. Cytoskeleton, as an intracellular load-bearing structure, plays an important role in gravity perception. Therefore, 13 cytoskeleton-related genes were chosen according to the results of microarray analysis, and the expressions of these genes were found to be altered under HMGE by real-time PCR. Based on the PCR results, the expressions of WASF2 (WAS protein family, member 2), WIPF1 (WAS/WASL interacting protein family, member 1), paxillin, and talin 1 were further identified by western blot assay. Results indicated that WASF2 and WIPF1 were more sensitive to altered gravity levels, and talin 1 and paxillin were sensitive to both magnetic field and gravity changes. Our findings demonstrated that HMGE can affect osteoblast gene expression profile and cytoskeleton-related genes expression. The identification of mechanosensitive genes may enhance our understandings to the mechanism of bone loss induced by microgravity and may provide some potential targets for preventing and treating bone loss or osteoporosis.

Protein-protein interaction network and significant gene analysis of osteoporosis

“…This result also supports a previous study (Qian et al, 2009), thus verifying the accuracy of the present study. Furthermore, we here identified the differentially expressed THFRSF1B, ESRRB, COL10A1, IBSP, TNFRSF11A, and WNT4 genes as potential key genes of osteoporosis.…”

“…Cell death had previously been shown to be related to osteoporosis (Bonewald, 2004), as mentioned above. The cytoskeleton is the bracing structure of the cell, and recent research has revealed that changes in the expression of cytoskeleton-related genes are closely associated with bone loss and osteoporosis (Qian et al, 2009).…”

Two-dimensional clinorotation influences cellular morphology, cytoskeleton and secretion of MLO-Y4 osteocyte-like cells

“…Our previous studies have shown that diamagnetic levitation caused by large gradient high superconducting magnet influences cytoskeleton arrangement and expression of associated genes in osteoblast-like and osteocyte-like cells (Qian et al 2008(Qian et al , 2009(Qian et al , 2010. In this study, 48 h of clinorotation, but not 12 h and 24 h of clinorotation, destroyed stress fibre formation and α-tubulin distribution in MLO-Y4 cells.…”

Mechano-biological Coupling of Cellular Responses to Microgravity

“…Both the height and roughness of cytoskeleton are attenuated (Qian et al 2010). Cytoskeleton-associated proteins are found to present altered expression in osteoblasts after being treated in HMGE, as seen in cDNA microarray analysis (Qian et al 2009a). The distributions of actin-binding proteins (vinculin, paxillin, zyxin, and talin) are dispersed and the expressions are also reduced consistently (Nabavia et al 2011;Qian et al 2012).…”