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ZFIN ID: ZDB-FIG-080909-12
Snow et al., 2008 - Muscle development is disrupted in zebrafish embryos deficient for fibronectin. Developmental dynamics : an official publication of the American Association of Anatomists   237(9):2542-2553 Full text @ Dev. Dyn.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: 14-19 somites to 20-25 somites
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: 14-19 somites to 20-25 somites

Fig. 1 A reduction in fibronectin (Fn) levels disrupts formation of anterior somites. A-D: Side views, anterior left, dorsal top, of 22-somite embryos stained for Fn (green) and β-catenin (blue) to visualize all cells. Somite 10, or the approximate region of somite 10, is to the left. A: In control embryos, Fn is concentrated at segment boundaries (white arrow). B: Injection of 6-ng fn1+3 morpholinos (MOs) results in a strong reduction Fn protein. C: In embryos injected with 3-ng fn1+3 MOs, Fn is reduced anteriorly (green arrow). Posteriorly, Fn concentration at boundaries is stronger, but not as strong as control posterior boundaries (white arrow). D: Quantification of levels of Fn in 3-ng and 6-ng morphants (y axis is arbitrary units). Fn protein is reduced in 3-ng fn1+3 morphants and strongly reduced in 6-ng fn1+3 morphants. E-G: Side views, anterior left, dorsal top, of 22-somite embryos stained with phalloidin to outline cells. E: Somite 7, or the approximate region of somite 7 is to the left. In control embryos, segment boundaries are clearly visible (white arrowheads). Segment boundaries were interactively traced (E1, see the Experimental Procudures section). The myotomes in control embryos are approximately the same size (E2). F: In 6-ng fn1+3 MO-injected embryos, anterior myotome boundary formation is severely disrupted (F, F1, white arrowheads indicate a tiny myotome boundary in the original data and the same image with myotome regions overlaid) and myotomes areas are large and irregular. G: Anterior myotomes are less disrupted in 3-ng fn1+3 MO-injected embryos (red arrowheads indicate where boundaries should be). H: Side views, anterior left, dorsal top, of 22-somite embryos stained with β-catenin, somite 17 or the approximate region thereof is on the left. In control embryos somite boundaries are seen as bright white lines (H, white arrow). In embryos injected with 6 ng of fn1+3 MOs imperfect posterior somite boundaries form (H1, white arrowheads). Robust somite boundaries form in the posterior of 3-ng fn1+3 morphant embryos (H2, white arrow). I-M: Dorsal views, anterior top, of 18-somite embryos. Analysis of mRNA expression correlates with the morphological analysis in that injection of 3-ng fn1+3 MOs disrupts anterior, but not posterior, patterning. Note that gene expression patterns in the posterior presomitic mesoderm such as papc and myf5 is segmental in both control and 3-ng fn1+3 MO-injected embryos (I, I2, J, J2, green arrowheads) but the segmental pattern is slightly disrupted in 6-ng fn1+3 MO-injected embryos (I1,J1, red arrowheads). myoD is expressed in stripes in control (K, green arrowhead) embryos throughout the A-P axis. Striped expression of myoD is disrupted in the anterior of 3 and 6-ng fn1+3 MO-injected embryos (K1, K2, red arrowheads) but is segmental posteriorly (K1, K2, green arrowheads). The expression patterns of myogenin (myog) and tropomyosin (tpm) are similar, but levels of myog and tpm in the posterior of 6-ng fn1+3 morphants are qualitatively reduced. Scale bar = 50 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
myf5 WT standard conditions 14-19 somites segmental plate ISH
14-19 somites somite ISH
WT + MO1-fn1a + MO1-fn1b + MO2-fn1b standard conditions 14-19 somites segmental plate ISH
WT + MO1-fn1a + MO1-fn1b + MO2-fn1b standard conditions 14-19 somites somite ISH
myod1 WT standard conditions 14-19 somites somite ISH
WT + MO1-fn1a + MO1-fn1b + MO2-fn1b standard conditions 14-19 somites somite ISH
myog WT standard conditions 14-19 somites somite ISH
WT + MO1-fn1a + MO1-fn1b + MO2-fn1b standard conditions 14-19 somites somite ISH
pcdh8 WT standard conditions 14-19 somites segmental plate ISH
14-19 somites somite ISH
WT + MO1-fn1a + MO1-fn1b + MO2-fn1b standard conditions 14-19 somites segmental plate ISH
14-19 somites somite ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-ctnnb IHC WT standard conditions 20-25 somites somite
IHC 20-25 somites trunk
IHC WT + MO1-fn1a + MO1-fn1b + MO2-fn1b standard conditions 20-25 somites somite
IHC 20-25 somites trunk
IHC WT + MO1-fn1a + MO1-fn1b + MO2-fn1b standard conditions 20-25 somites somite
IHC 20-25 somites trunk
Ab1-fn IHC WT standard conditions 20-25 somites somite border
IHC 20-25 somites trunk
IHC WT + MO1-fn1a + MO1-fn1b + MO2-fn1b standard conditions 20-25 somites somite border
IHC 20-25 somites trunk
IHC WT + MO1-fn1a + MO1-fn1b + MO2-fn1b standard conditions 20-25 somites somite border
IHC 20-25 somites trunk
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-fn1a + MO1-fn1b + MO2-fn1b standard conditions 14-19 somites somite shape, abnormal
14-19 somites somite border aplastic, abnormal
14-19 somites somite border shape, abnormal
14-19 somites somitogenesis disrupted, abnormal
20-25 somites myotome increased size, abnormal
20-25 somites myotome variant shape, abnormal
20-25 somites somite shape, abnormal
20-25 somites somite border aplastic, abnormal
20-25 somites somite border shape, abnormal
20-25 somites somitogenesis disrupted, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental dynamics : an official publication of the American Association of Anatomists for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Dev. Dyn.