ZFIN ID: ZDB-FIG-090717-2
Lin et al., 2009 - The transcription factor six1a plays an essential role in the craniofacial myogenesis of zebrafish. Developmental Biology   331(2):152-166 Full text @ Dev. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Prim-25 to Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Long-pec to Protruding-mouth

Fig. 2 Six1a is required for the development of mr, ir, sh and all pharyngeal muscles. Embryos derived from the transgenic line Tg(α-actin:RFP) (A–H), all of whose skeletal muscles appear as red fluorescent protein (RFP), were injected with 8 ng of six1a-morpholino oligonucleotide (MO) to specifically inhibit six1a mRNA translation. RFP signal was detected only in the so, io, sr, lr, and remnant dorsal branchial arch muscle (white star) primordia in the six1a-MO-injected embryos (A vs. C and B vs. D). When embryos were injected with 8 ng of six1-MO together with 150 pg of six1a mRNA, results showed that the defective muscle primordia induced by six1a-MO were rescued and appeared as RFP-labeled muscles (E, F; the rescued muscles are marked in green typeface). In contrast, the rescue experiment failed when embryos were injected with 8 ng of six1a-MO with 200 pg of gfp mRNA (G, H), suggesting that the defects of six1a morphants were specific. The expressions of myf5 (I, J, M, N), myod (K, O), and myogenin (L, P) were also observed at the stages indicated. When wild-type embryos were injected with six1a-MO, myf5 was expressed normally in the six1a morphants, both at 36- (I vs. M) and at 48-hpf (J vs. N), except sh. On the other hand, the expressions of myod (K vs. O) and myogenin (L vs. P) were decreased in the extraocular io, so, sr and lr in the six1 morphants at 48 hpf. Weak signals of myod and myogenin were also noticed in the remnant dorsal branchial muscles (black stars) of six1a morphants. The schematic diagram illustrates the cranial muscle defects in six1a morphants and compares the expressions of myf5, myod and myogenin between wild-type (upper row, Q) and six1a morphants (lower row, Q). Lateral view: A, C, E, G and I–P; and ventral view: B, D, F and H. ah, adductor hyoideus; am, adductor mandibulae; ao, adductor operculi; do, dilator operculi; hh, hyohyoideus; ih, interhyoideus; ima, intermandibularis anterior; imp, intermandibularis posterior; io, inferior oblique; ir, inferior rectus; lap, levator arcus palatini; lr, lateral rectus; mr, medial rectus; sh, sternohyoideus; so, superior oblique and sr, superior rectus.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
myf5 WT control Prim-25 dorsal oblique extraocular muscle ISH
Prim-25 pharyngeal arch 2 ISH
Prim-25 pharyngeal arch 3 ISH
Prim-25 ventral oblique extraocular muscle ISH
Long-pec dorsal oblique extraocular muscle ISH
Long-pec pharyngeal arch 3 ISH
Long-pec sternohyoid ISH
Long-pec ventral oblique extraocular muscle ISH
WT + MO1-six1b standard conditions Prim-25 dorsal oblique extraocular muscle ISH
Prim-25 pharyngeal arch 2 ISH
Prim-25 pharyngeal arch 3 ISH
Prim-25 ventral oblique extraocular muscle ISH
Long-pec dorsal oblique extraocular muscle ISH
Long-pec pharyngeal arch 3 ISH
Long-pec Not Detected sternohyoid ISH
Long-pec ventral oblique extraocular muscle ISH
myod1 WT control Long-pec adductor hyohyoid ISH
Long-pec adductor mandibulae ISH
Long-pec constrictor dorsalis ISH
Long-pec dorsal oblique extraocular muscle ISH
Long-pec dorsal rectus ISH
Long-pec hyohyoideus ISH
Long-pec lateral rectus ISH
Long-pec mandibular muscle ISH
Long-pec medial rectus ISH
Long-pec sternohyoid ISH
Long-pec ventral oblique extraocular muscle ISH
Long-pec ventral rectus ISH
WT + MO1-six1b standard conditions Long-pec dorsal oblique extraocular muscle ISH
Long-pec dorsal rectus ISH
Long-pec lateral rectus ISH
Long-pec ventral oblique extraocular muscle ISH
myog WT control Long-pec adductor hyohyoid ISH
Long-pec adductor mandibulae ISH
Long-pec constrictor dorsalis ISH
Long-pec dorsal oblique extraocular muscle ISH
Long-pec dorsal rectus ISH
Long-pec hyohyoideus ISH
Long-pec lateral rectus ISH
Long-pec mandibular muscle ISH
Long-pec medial rectus ISH
Long-pec sternohyoid ISH
Long-pec ventral oblique extraocular muscle ISH
Long-pec ventral rectus ISH
WT + MO1-six1b standard conditions Long-pec dorsal oblique extraocular muscle ISH
Long-pec dorsal rectus ISH
Long-pec lateral rectus ISH
Long-pec ventral oblique extraocular muscle ISH
RFP twu3Tg control Protruding-mouth adductor hyohyoid IFL
Protruding-mouth adductor mandibulae IFL
Protruding-mouth adductor operculi IFL
Protruding-mouth dilatator operculi IFL
Protruding-mouth dorsal oblique extraocular muscle IFL
Protruding-mouth dorsal rectus IFL
Protruding-mouth hyohyoideus IFL
Protruding-mouth lateral rectus IFL
Protruding-mouth levator arcus palatini IFL
Protruding-mouth mandibular muscle IFL
Protruding-mouth sternohyoid IFL
Protruding-mouth ventral oblique extraocular muscle IFL
Protruding-mouth ventral rectus IFL
twu3Tg + MO1-six1b standard conditions Protruding-mouth dorsal oblique extraocular muscle IFL
Protruding-mouth dorsal rectus IFL
Protruding-mouth lateral rectus IFL
Protruding-mouth ventral oblique extraocular muscle IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-six1b standard conditions Long-pec medial rectus aplastic, abnormal
Long-pec ventral rectus aplastic, abnormal
twu3Tg + MO1-six1b standard conditions Protruding-mouth dorsal oblique extraocular muscle present, normal
Protruding-mouth dorsal rectus present, normal
Protruding-mouth lateral rectus present, normal
Protruding-mouth medial rectus aplastic, abnormal
Protruding-mouth pharyngeal arch muscle aplastic, abnormal
Protruding-mouth sternohyoid aplastic, abnormal
Protruding-mouth ventral oblique extraocular muscle present, normal
Protruding-mouth ventral rectus aplastic, abnormal
Acknowledgments:
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Reprinted from Developmental Biology, 331(2), Lin, C.Y., Chen, W.T., Lee, H.C., Yang, P.H., Yang, H.J., and Tsai, H.J., The transcription factor six1a plays an essential role in the craniofacial myogenesis of zebrafish, 152-166, Copyright (2009) with permission from Elsevier. Full text @ Dev. Biol.