21.

3 Specification and Differentiation of Muscle

Embryonic Somites Give Rise to Myoblasts

Specific signals from surrounding tissues play an important role in determining where myoblasts will form As myoblasts proliferate and migrate, they become aligned, stop dividing, and fuse to form syncytium Cell containing many nuclei but sharing cytoplasm (myotube)

Myogenic Genes were First Identified in Studies with Cultured Myoblasts

C3H 10T1/2
Central role in dissecting transcription control mechanisms regulating skeletal myogenesis

Myogenic Genes were First Identified in Studies with Cultured Myoblasts

myoD cDNA
Myogenic determination gene C3H 10T1/2 transfected with myoD cDNA and treated with 5azacytidine both formed myotubes myoD gene proposed to play a key role in muscle development Same approach to myogenin, myf5, mrf4

Regulatory Factors in Guiding Production of Muscle Cells

BHLH: Basic HelixLoop-Helix MRF
Produced only in developing muscles B region: mediates dimer formation

Associate with E2A and MEF to form large transcriptional complexes that drive myogenesis and expression of musclespecific genes

Regulatory Factors in Guiding Production of Muscle Cells

MEF Importance:
Many musclespecific genes contain recognition sites for both MRF and MEF Enhance ability of MRF o induce myogenic conversion

Differentiation of Myoblasts is Under Positive and Negative Control

Production of muscle regulators is regulated only in mesoderm cells in response to locally acting signals Chromatin remodeling proteins are needed to target genes accessible to MRFs Inhibitory proteins can restrict MRFs act Antagonistic relations between cell-cycle regulators and differentiation factors like MRFs ensure that differentiating cells will not divide, v.v.

Activating Chromatin-Remodeling Proteins

Carried out by large protein complexes (SWI/SWF complex) that have ATPase and perhaps helicase activity
Recruit histone acetylases that modify chromatin to make genes accessible to transcription factors

Transcription activators by myogenic proteins depend on a suitable chromatin structure in the regions of muscle-specific genes
MEF2 recruits histone acetylases through mediator -> activate

Inhibitory Proteins
Myogenic program is inhibited by:
Binding Id protein to MyoD
Blocks binding of MyoD to DNA Id prevents cells that produce MyoD and E2A from activating transcription of the muscle-specific gene encoding creatine kinase
Remain in proliferative state

Histone deacetylases
Repress activation of target genes by MRFs MEF2, through MADS domain, can bind to a histone deacetylase which can prevent MEF2 function and muscle differentiation
Histone deacetylase phosphorylation by CA2+/calmodulin-dependent protein kinase, then is moved from nucleus to cytoplasm

Cell-Cycle Proteins
Cyclins and CDKs
Influences determined to differentiated state transition

Differentiation of cultured myoblasts can be inhibited by transfecting cells with DNA encoding cyclin D1
Mimic antagonization of differentiation pathway via in vivo signals

Cell-Cell Signals are Crucial for Determination and Migration of Myoblasts

Pax3
Control muscle formation in the body all and limbs

Lbx1
Produced by myoblasts that will migrate Dependent on Pax3

SF/HGF
Scatter factor/Hepatocyte growth factor Attracts migrating myoblasts, directing them to proper destination

bHLH Regulatory Proteins Function in Creation of Other Tissues