ZFIN ID: ZDB-FIG-110317-18
Osborn et al., 2011 - Cdkn1c drives muscle differentiation through a positive feedback loop with Myod. Developmental Biology   350(2):464-475 Full text @ Dev. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Prim-5 to Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Prim-5 to Protruding-mouth

Fig. 6 Cdkn1c drives fast myogenesis through Myod. In situ mRNA hybridization (A,D) or protein immunodetection (B,C) at 24 hpf (A–C; lateral flatmount) or 72 hpf (D, dorsal flatmount) of embryos injected with cdkn1c MO. (A). Cdkn1c MO exacerbated somitic fast muscle loss induced by myod MO, and ablated residual fast muscle in mutant myf5hu2022;myod knockdown. Bar: 100 μm (B). Cdkn1c MO alone diminishes fast muscle differentiation, without reducing slow muscle. Dashed lines indicate positions of upper and lower transverse sections. Bars: flatmounts 100 μm, somite lateral zoom and sections 50 μm (C). Cdkn1c MO diminished Myod immunoreactivity in nuclei of nascent fast muscle and Myogenin immunoreactivity in more mature somites, particularly the expression at the posterior somite border (arrows). Bar: 50 μm. (D). Cdkn1c MO (9 ng) reduced ventral head myogenesis in a similar manner to myod MO (2 ng). Low doses of cdkn1c MO (3 ng) or myod MO (0.25 ng) that alone had little effect, synergised to reduce myogenesis in intermandibular, interhyoideus and hyohoideus muscles. Bar: 100 μm. Chart quantifies the extent of ventral hyoid muscle defects after injection of the indicated ng of each MO/embryo, with differences tested by χ2 test. All embryos were classified as having either ‘normal’, ‘weak’ or ‘no’ mylz2 mRNA in the three stated muscles. ‘Normal’ embryos (dark bars) had strong mylz2 expression, as shown in control panel. ‘Weak’ embryos (light bars) had mylz2 expression visibly below the control range, as shown in the 2 ng myod MO panel. ‘No’ embryos entirely lacked mylz2 expression, as in the 9 ng cdkn1c MO panel, are shown as white space above the bars. Data shown has n = 20 for each condition from within a single experiment.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
myhz1.1 WT control Prim-5 fast muscle cell ISH
WT + MO1-myod1 standard conditions Prim-5 fast muscle cell ISH
WT + MO1-myod1 + MO3-cdkn1ca standard conditions Prim-5 fast muscle cell ISH
myf5hu2022/hu2022 + MO1-myod1 standard conditions Prim-5 fast muscle cell ISH
myf5hu2022/hu2022 + MO1-myod1 + MO3-cdkn1ca standard conditions Prim-5 Not Detected fast muscle cell ISH
mylpfa WT control Protruding-mouth head muscle ISH
Protruding-mouth hyohyoideus ISH
Protruding-mouth interhyoideus ISH
WT + MO1-myod1 standard conditions Protruding-mouth head muscle ISH
Protruding-mouth hyohyoideus ISH
Protruding-mouth interhyoideus ISH
WT + MO1-myod1 + MO3-cdkn1ca standard conditions Protruding-mouth head muscle ISH
Protruding-mouth Not Detected hyohyoideus ISH
Protruding-mouth Not Detected interhyoideus ISH
WT + MO3-cdkn1ca standard conditions Protruding-mouth head muscle ISH
Protruding-mouth hyohyoideus ISH
Protruding-mouth interhyoideus ISH
myod1 Ab1-myod1 WT control Prim-5 fast muscle cell nucleus IHC
Ab1-myod1 WT + MO3-cdkn1ca standard conditions Prim-5 fast muscle cell nucleus IHC
myog WT control Prim-5 somite IHC
Prim-5 somite border IHC
WT + MO3-cdkn1ca standard conditions Prim-5 Not Detected somite border IHC
Prim-5 tail bud IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab-A4.1025 IHC WT control Prim-5 fast muscle cell
IHC Prim-5 slow muscle cell
IHC WT + MO3-cdkn1ca standard conditions Prim-5 fast muscle cell
IHC Prim-5 slow muscle cell
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-myod1 standard conditions Prim-5 fast muscle cell decreased amount, abnormal
Protruding-mouth head muscle hypoplastic, abnormal
WT + MO1-myod1 + MO3-cdkn1ca standard conditions Prim-5 fast muscle cell decreased amount, abnormal
Protruding-mouth head muscle hypoplastic, abnormal
WT + MO3-cdkn1ca standard conditions Prim-5 fast muscle cell decreased amount, abnormal
WT + MO3-cdkn1ca standard conditions Prim-5 slow muscle cell normal amount, normal
WT + MO3-cdkn1ca standard conditions Protruding-mouth head muscle hypoplastic, abnormal
myf5hu2022/hu2022 + MO1-myod1 standard conditions Prim-5 fast muscle cell decreased amount, abnormal
myf5hu2022/hu2022 + MO1-myod1 + MO3-cdkn1ca standard conditions Prim-5 fast muscle cell absent, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Biology for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Biology, 350(2), Osborn, D.P., Li, K., Hinits, Y., and Hughes, S.M., Cdkn1c drives muscle differentiation through a positive feedback loop with Myod, 464-475, Copyright (2011) with permission from Elsevier. Full text @ Dev. Biol.