Biochemical and Biophysical Research Communications
Regulation of sprouty expression by PLCγ and calcium-dependent signals
Section snippets
Materials and methods
Cell culture conditions. ATDC5 cells were maintained in a log growth phase in DMEM/Ham’s F12 hybrid medium containing 5% (v/v) FBS (Hyclone), 10 μg/ml human transferrin (Calbiochem), and 3 × 10−8 M sodium selenite (Sigma) at 37 °C in a humidified 5% CO2/95% air atmosphere. In the sprouty induction study, we plated ATDC5 cells in six-multiwell plastic plates (Falcon) at an initial density of 30 × 104 cells/well and cultured these cells without changing the medium for 48 h to obtain quiescent state. For
bFGF induces sprouty1 and sprouty2 in ATDC5 cells
The mouse chondrogenic cell line, ATDC5, was stimulated with bFGF and Spry1 and Spry2 gene expression was assessed by RT-PCR (Fig. 1). Expression of both genes was induced after 1 h of bFGF addition. Spry1 expression achieved maximal expression 4 h after stimulation with bFGF and elevated levels of sprouty expression were maintained for at least 12 h. Similarly, Spry2 was induced 1 h after addition of bFGF and high expression continued at least 12 h after bFGF addition. No amplification product was
Discussion
Others [15] and we have shown that erk MAP kinases contribute to FGF-induced sprouty gene expression. However, a requirement for alternative pathways to induce sprouty expression is suggested by the finding that erk MAP kinases are repressed in certain tissues where sproutys are expressed. For example, in limb bud mesenchyme erk MAP kinases are largely inactive [16], [17], yet sproutys are strongly expressed. Therefore, in this report we investigated if calcium-dependent pathways regulate
Acknowledgments
We thank Victoria Frohlich and James Lechleiter for assistance with confocal imaging and analysis. This work was supported by NIH Grants AR47070, AR050024 and the Paul Beeson Physician Faculty Scholars in Aging Research Program.
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2015, International Journal of Biochemistry and Cell BiologyCitation Excerpt :Accordingly, Spry protein expression is induced by growth factors as a consequence of mitogen-activated protein kinase (MAPK) activation (Gross et al., 2001; Ozaki et al., 2001). In addition, their expression can be influenced by calcium release (Abe and Naski, 2004) and hypoxia (Haigl et al., 2010). In an earlier study, we investigated Spry2 and Spry4 expression throughout the cell cycle and found that the Spry proteins show a specific cell cycle distribution, although both Spry proteins are induced when serum-arrested cells were released (Mayer et al., 2010).
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2011, Journal of Molecular BiologyCitation Excerpt :They exhibit partial overlap of the expression patterns8 and the biochemical properties.9–11 They have been shown to function as general inhibitors of receptor tyrosine kinases (RTKs) 12–14 and are induced in response to RTK signaling through both MAP kinase15 and calcium signaling pathways.16 The molecular mechanisms by which Spry2 might inhibit RTK or Ca2+signaling are uncertain but it is generally believed to be through protein–protein interactions with its multiple binding partners.10,11
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P2Y receptors activated by diadenosine polyphosphates reestablish Ca<sup>2+</sup> transients in achondroplasic chondrocytes
2008, BoneCitation Excerpt :After determining the existence and functionality of P2Y receptors in achondroplasic chondrocytes, the other main objective of this work has been to analyze the possible effect of activated P2Y receptors on the lack of Ca2+ responses in FGF9-stimulated achondroplasic chondrocytes. Some reports have described the increase of intracellular calcium levels induced by wild-type FGFR3 in response to FGF exposure [4,20]. Binding of FGF to FGFR3 leads to autophosphorylation of the receptor on several tyrosine residues, being the phosphorylation of Tyr760 essential for interaction and subsequent activation of PLCγ.
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