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Yan Zhang, Wendy Yang, Abraham Hang, Emilia Zin, Mariana Garcia, Mei Li, Christine Frances Wildsoet; BMP2 Protein Increases the Expression of Genes for Inhibitor of DNA Binding Proteins in Cultured Chick Scleral Fibroblasts. Invest. Ophthalmol. Vis. Sci. 2017;58(8):5472. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
The Bone Morphogenetic Protein (BMP) signaling pathway has previously been linked to defocus-driven eye growth regulation in chick. This study used cultured chick scleral fibroblasts as an in vitro model system to investigate the effects of recombinant human BMP2 (rhBMP2) protein on elements of this signaling pathway.
Chick scleral fibroblasts (CSFs) were cultured in DMEM/F12 medium with 10% FBS. The CSFs were treated with 0, 1, 10, 100, or 1000 ng/ml BMP2 in DMEM/F12 medium for two hours, after which cells were collected and RNA extracted, purified and transcribed into cDNA. Gene expression of BMP2, BMP4, BMP7, BMPR1A, BMPR1B, BMPR2, SMAD1, SMAD5, SMAD9, ID1-4 was quantified with qPCR and protein expression of the same signaling molecules examined with Western blot and immunocytochemistry.
Genes for all of the above signaling molecules were detected, as were the SMAD1, p-SMAD1, and p-SMAD1/5 proteins. With all applied concentrations of rhBMP2, the gene expression of Inhibitor of DNA Binding proteins (ID1-4), were significantly increased (Table 1).
The BMP signaling pathway is very likely involved in chick scleral remodeling, as the exposure of cultured CSF to rhBMP2 resulted in significant changes in gene expression of down-stream transcription factors, i.e., Inhibitor of DNA Binding proteins.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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