ZFIN ID: ZDB-FIG-090223-20
Hinits et al., 2009 - Differential requirements for myogenic regulatory factors distinguish medial and lateral somitic, cranial and fin muscle fibre populations. Development (Cambridge, England)   136(3):403-414 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: 10-13 somites to Prim-5

Fig. 4 Myf5 or Myod is required for medial fast myogenesis. In situ hybridisation for myog, myhz1, tpma, mylz2 or mef2d or immunohistochemistry for the indicated myosins in wild-type or myf5+/hu2022 incross zebrafish injected with the indicated MOs. Lateral views, anterior to left unless otherwise stated. (A) myog expression was delayed in embryos injected with myf5+myod MOs. Dorsolateral views, anterior to top. (B) MyHC was reduced in both epaxial and hypaxial domains in myod MO embryos, grossly reduced in double myf5+myod morphants and further diminished in myf5hu2022 mutants injected with myod MO. Note the complete absence of residual cells in some individual somites (dotted outline). (C) Residual differentiated muscle in myf5+myod morphants expressed fast but not slow MyHC at 24 hpf. Lateral view of confocal stack of somites 9-10. (D) Fast muscle markers were indistinguishable in 24 hpf myf5hu2022 homozygous mutants and their siblings (left column), reduced by myod MO in siblings (middle) and almost eliminated by myod MO in myf5hu2022 mutants (right). Inset illustrates that myf5 MO does not worsen the myod MO plus myf5hu2022 phenotype. mylz2 mRNA accumulation (red, arrowheads) and myog downregulation (blue) are delayed at ∼20 ss in fast muscle. (Bottom panels) Transverse sections of double knockdowns show residual fast muscle (arrowheads) immediately lateral to adaxial and/or twist2-expressing sclerotomal cells (arrows). (E) (Left, top) mef2d mRNA initiates normally in pbcs in the myod morphant, but is restricted to the medial region of rostral somites (yellow brackets). In myf5+myod MOs, mef2d mRNA is absent. Dorsal flatmounts, anterior to top. (Left, bottom) Lack of both myf5 and myod ablates mef2d expression. Lateral views, anterior to top. (Right) By 22 ss, transverse sections (at the level of the bar on lateral flatmounts) show reduction of mef2d mRNA (purple) in myod or myog, but not in myf5, single morphants. Mef2 position is shown relative to slow fibres (brown). Note the expression of mef2d lateral to migrating slow fibres in the control and in myf5 and myog, but not myod, morphants (arrows).

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
mef2d WT control 14-19 somites adaxial cell ISH
14-19 somites somite ISH
20-25 somites adaxial cell ISH
20-25 somites somite ISH
WT + MO1-myod1 standard conditions 14-19 somites adaxial cell ISH
14-19 somites somite ISH
20-25 somites adaxial cell ISH
20-25 somites somite ISH
WT + MO1-myod1 + MO3-myf5 standard conditions 14-19 somites Not Detected adaxial cell ISH
14-19 somites Not Detected somite ISH
WT + MO1-myog standard conditions 20-25 somites adaxial cell ISH
20-25 somites somite ISH
WT + MO3-myf5 standard conditions 20-25 somites adaxial cell ISH
20-25 somites somite ISH
myf5hu2022/hu2022 standard conditions 14-19 somites adaxial cell ISH
14-19 somites somite ISH
myf5hu2022/hu2022 + MO1-myod1 standard conditions 14-19 somites Not Detected adaxial cell ISH
14-19 somites Not Detected somite ISH
myhz1.1 WT + MO1-myod1 standard conditions Prim-5 somite ISH
myf5hu2022/hu2022 standard conditions Prim-5 somite ISH
myf5hu2022/hu2022 + MO1-myod1 standard conditions Prim-5 somite ISH
myf5hu2022/hu2022 + MO1-myod1 + MO3-myf5 standard conditions Prim-5 somite ISH
mylpfa myf5hu2022/hu2022 standard conditions 20-25 somites fast muscle cell ISH
Prim-5 fast muscle cell ISH
myf5hu2022/hu2022 + MO1-myod1 standard conditions 20-25 somites fast muscle cell ISH
Prim-5 fast muscle cell ISH
myog WT control 10-13 somites adaxial cell ISH
10-13 somites somite ISH
20-25 somites adaxial cell ISH
20-25 somites somite ISH
WT + MO1-myod1 + MO3-myf5 standard conditions 10-13 somites Not Detected adaxial cell ISH
10-13 somites Not Detected somite ISH
20-25 somites adaxial cell ISH
20-25 somites somite ISH
tpma WT + MO1-myod1 standard conditions Prim-5 somite ISH
myf5hu2022/hu2022 standard conditions Prim-5 somite ISH
myf5hu2022/hu2022 + MO1-myod1 standard conditions Prim-5 somite ISH
twist2 WT + MO1-myod1 standard conditions 20-25 somites to Prim-5 sclerotome ISH
myf5hu2022/hu2022 + MO1-myod1 standard conditions 20-25 somites to Prim-5 sclerotome ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab-A4.1025 IHC WT control Prim-5 epaxial myotome region
IHC Prim-5 hypaxial myotome region
IHC WT + MO1-myod1 standard conditions Prim-5 epaxial myotome region
IHC Prim-5 hypaxial myotome region
IHC WT + MO1-myod1 + MO3-myf5 standard conditions Prim-5 epaxial myotome region
IHC Prim-5 hypaxial myotome region
IHC myf5hu2022/hu2022 standard conditions Prim-5 epaxial myotome region
IHC Prim-5 hypaxial myotome region
IHC myf5hu2022/hu2022 + MO1-myod1 standard conditions Prim-5 epaxial myotome region
IHC Prim-5 hypaxial myotome region
Ab-EB165 IHC WT control Prim-5 fast muscle cell
IHC WT + MO1-myod1 + MO3-myf5 standard conditions Prim-5 fast muscle cell
Phenotype Details No data available
Acknowledgments:
ZFIN wishes to thank the journal Development (Cambridge, England) for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Development