About: c-Myb contributes to endochondral bone development     Goto   Sponge   NotDistinct   Permalink

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  • The expression of c-Myb was analyzed in fore- and hindlimbs of mouse embryos during early embryonic stages using in situ hybridization. c-MYB protein was detected throughout prenatal development using immunohistochemistry. In the limb, weak expression of c-myb was observed in embryos at E12.5. At later stages (E13.5 and E14.5), expression was located in the perichondrium and interdigital areas, which correlated with localization of c-MYB protein. Furthermore, we detected the c-MYB protein in pre-hypertrophic and hypertrophic chondrocytes. Transient overexpression of c-myb by transfection of a c-Myb-coding vector markedly increased the formation of cartilage nodules and the production of extracellular matrix, as detected by intense staining with Alcian Blue. Moreover, the expression of Sox9, a major marker of chondrogenesis, was increased. A loss-of-function approach using c-myb specific siRNA decreased nodule formation, as well as downregulating the level of Sox9 expression. Furthermore, the c-Myb significantly enhanced the activity of a Col2-LUC reporter gene compared to a control vector. Additional gene expression changes were analyzed by PCR Array designed for osteogenic markers. Overexpression of c-myb using micromass cultures following 24 hour of incubation significantly increased expression of several cartilage and bone markers such as Bmpr1a, Smad1, Msx1, Col1a2, Fgfr2, Tgfb3, Icam1. In addition a strong increase in the expression of NF-kappab1 and Vegfa was evident at this early time point. NF-kappab is known to control the differentiation and activity of the major skeletal cell types - osteoclasts, osteoblasts, osteocytes and chondrocytes (Novack, 2011), while Vegfa is necessary for chondrocyte survival during bone development (Zelzer et al., 2003). In conclusion, our results show that the transcription factor c-Myb plays a role in promotion and regulation of endochondral bone formation.
  • The expression of c-Myb was analyzed in fore- and hindlimbs of mouse embryos during early embryonic stages using in situ hybridization. c-MYB protein was detected throughout prenatal development using immunohistochemistry. In the limb, weak expression of c-myb was observed in embryos at E12.5. At later stages (E13.5 and E14.5), expression was located in the perichondrium and interdigital areas, which correlated with localization of c-MYB protein. Furthermore, we detected the c-MYB protein in pre-hypertrophic and hypertrophic chondrocytes. Transient overexpression of c-myb by transfection of a c-Myb-coding vector markedly increased the formation of cartilage nodules and the production of extracellular matrix, as detected by intense staining with Alcian Blue. Moreover, the expression of Sox9, a major marker of chondrogenesis, was increased. A loss-of-function approach using c-myb specific siRNA decreased nodule formation, as well as downregulating the level of Sox9 expression. Furthermore, the c-Myb significantly enhanced the activity of a Col2-LUC reporter gene compared to a control vector. Additional gene expression changes were analyzed by PCR Array designed for osteogenic markers. Overexpression of c-myb using micromass cultures following 24 hour of incubation significantly increased expression of several cartilage and bone markers such as Bmpr1a, Smad1, Msx1, Col1a2, Fgfr2, Tgfb3, Icam1. In addition a strong increase in the expression of NF-kappab1 and Vegfa was evident at this early time point. NF-kappab is known to control the differentiation and activity of the major skeletal cell types - osteoclasts, osteoblasts, osteocytes and chondrocytes (Novack, 2011), while Vegfa is necessary for chondrocyte survival during bone development (Zelzer et al., 2003). In conclusion, our results show that the transcription factor c-Myb plays a role in promotion and regulation of endochondral bone formation. (en)
Title
  • c-Myb contributes to endochondral bone development
  • c-Myb contributes to endochondral bone development (en)
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  • c-Myb contributes to endochondral bone development
  • c-Myb contributes to endochondral bone development (en)
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  • RIV/62157124:16170/14:43873326!RIV15-MSM-16170___
http://linked.open...avai/riv/aktivita
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  • 7609
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  • RIV/62157124:16170/14:43873326
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  • chondrogenic markers; chondrogenesis; c-Myb (en)
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  • [064BFF72B9E2]
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  • Buchtová, Marcela
  • Knopfová, Lucia
  • Šmarda, Jan
  • Janečková, Eva
  • Matalová, Eva
  • Šnajdr, Pavel
  • Švandová, Eva
  • Oralová, Veronika
  • Krejčí, Eliška
  • Tucker, Abigail
http://localhost/t...ganizacniJednotka
  • 16170
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