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Protein kinase A signalling via CREB controls myogenesis induced by Wnt proteins

Protein kinase A signalling via CREB controls myogenesis induced by Wnt proteins. Chen et al , 2005, Nature. Myogenesis. (D). (V). Sclerotome → vertebrae, ribs Myotome → skeletal muscles Medial myotome → dorsal trunk muscles Lateral myotome → ventral limb muscles

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Protein kinase A signalling via CREB controls myogenesis induced by Wnt proteins

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  1. Protein kinase A signalling via CREB controls myogenesis induced by Wnt proteins Chen et al, 2005, Nature

  2. Myogenesis (D) (V) Sclerotome → vertebrae, ribs Myotome → skeletal muscles Medial myotome→ dorsal trunk muscles Lateral myotome → ventral limb muscles Dermatome→ dermis (skin) Pax3 www.google.com

  3. Wnt & Myogenesis The EMBO Journal, review, 1999

  4. The EMBO Journal, review, 1999 Delivery of an activated form of beta-catenin to somitic mesoderm mimics the effects of Wnt-1, demonstrating that Wnt-1 likely acts directly on somitic mesoderm, and not through adjacent tissues via an indirect signal relay mechanism. (Capdevila et al, 1998, Dev Biol.)

  5. CREB signaling PKA : cAMP-dependent Protein Kinase A CRE : cAMP Responsive Element CREB : CRE Binding protein (basic leucine zipper protein)

  6. E 9.5 day Immunohistochemistry Psm : Presomitic mesoderm sI : first forming somite sV : fifth forming somite nt : neural tube dsom : dorsal somitic cells scl : sclerotome

  7. Psm : Presomitic mesoderm tb : tail bud. E9.5 mouse embryo.

  8. drg : dorsal root ganglion nc : notochord my : myotome dm : dermomyotome E 10.5 day

  9. Shh Pax1 Whole-mount In Situ Hybridization CREB null mutant : Wnt1-/-, Wnt3a-/- embroys

  10. Immunohistochemistry Northern analysis ATF-1 (activating transcription factor-1), CREM (CRE modulator) : CREB family members

  11. ACREB(acidic-CREB) : dominant-negative inhibitor of all CREB family members Microinjection : GFP(control) & ACREB-expressing adenovirus fl : forelimb

  12. co-culture : psm(presomitic mesoderm) with B1a, Wnt1&7a-B1a RT-PCR ACREB : inhibitor of CREB C : positive control cDNA from E10.5 embryos Actb(ß-actin) : normalization control Canonical wnt target gene

  13. wnt P CREB CRE Luc Immunohistochemistry : P-CREB CRE-Luc reporter assay 10T1/2 : myogenic potential C57MG : Wnt-responsive ddA : adenylyl cyclase inhibitor dnP : dominant-negative form of PKA psm cells P-CREB staining

  14. co-culture : psm/ B1a → RT-PCR CTX, PTX : AC stimulator M7(Mas7) : Gαi(inhibitory subunit) stimulator M17(Mas17) : M7 inactive analogue ddA : AC(adenylyl cyclase) inhibitor fsk(forskolin) : AC activator dbc : cAMP activator TPA : PKC activator dnP : PKA inhibitor

  15. Microinjection (in vivo) PKA inhibitor PKA activity is required for Wnt-mediated myogenesis in vitro & in vivo

  16. cf) ß-catenin Cytoplasmic localization cAMP accumulation PKA activation CREB phosphorylation WISP-1 transcription ↑ (Wnt-1 induced secreted protein) Lifeng Xu et al, 2000, Genes & Dev Discussion Pourquié et al, 2005, nature <news and views>

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