1 / 17

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

hiero
Download Presentation

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

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  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>

More Related