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ZFIN ID: ZDB-FIG-131029-10
Neugebauer et al., 2013 - Differential roles for 3-OSTs in the regulation of cilia length and motility. Development (Cambridge, England)   140(18):3892-3902 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Prim-5 to Long-pec
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Long-pec

Fig. 1

3-OST-5 is required in whole embryos and cell autonomously in Kupffer’s vesicle for LR development. (A,B) Differential interference contrast images of 48 hpf embryos. Uninjected (A) and 3-OST-5 MO-injected (B) embryos at 48 hpf showing dose-dependent curly down tail phenotype. Both 3-OST-5 MOs showed a dose-dependent increase in both heart and gut reversals. (C,D) cmlc2-gfp transgenic fish (expressing GFP in cardiomyocytes) were used to visualize normal left looping (C) and reversed right looping (D) in 3-OST-5 MO1 embryos at 48 hpf. (E) Percentages of reversed heart and gut looping in 3-OST-5 morphants. Injection of 3-OST-5 MO1 (n=120; **P<0.001) or 3-OST-5 MO2 (n=121; **P<0.001) resulted in significant heart and gut looping compared with uninjected embryos (n=361). Co-injection of 3-OST-5 MO1 and 3-OST-5 ‘rescue’ mRNA resulted in a significant rescue of heart and gut orientation (n=137; **P<0.001). Injection of 3-OST-5 ‘rescue’ mRNA alone (n=171) or 3-OST-3Z MO (n=106) did not alter normal heart and gut looping. Error bars represent s.d. for all heart and gut looping treatments. (F) Representative images of normal (left-sided), reversed (right-sided), absent and bilateral spaw expression in 3-OST-5 morphants. Percentages of normal (left-sided), reversed (right-sided), absent, and bilateral spaw expression in the LPM of 3-OST-5 morphants (MO1, n=183; MO2, n=206), 3-OST-3Z morphants (n=240) or uninjected control embryos (n=220). 3-OST-5 morphants had a preponderance of bilateral expression, in comparison with predominantly left-sided expression in uninjected and 3-OST-3Z MO1-injected embryos. Embryos injected for DFC/KV-targeted knockdown, DFC3-OST-5 MO1 (n=112), showed a high rate of bilateral spaw expression. This is in contrast to DFCcontrol MO (n=97), Yolk3-OST-5 MO1 (n=61) and Yolkcontrol MO (n=90) injected embryos showing left-sided spaw expression.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
EGFP twu34Tg + MO1-hs3st1l2 standard conditions Long-pec cardiac muscle cell IFL
spaw WT standard conditions Prim-5 to Long-pec lateral plate mesoderm left side IHC
WT + MO1-hs3st1l2 standard conditions Prim-5 to Long-pec Not Detected lateral plate mesoderm IHC
Prim-5 to Long-pec lateral plate mesoderm lateral side IHC
Prim-5 to Long-pec lateral plate mesoderm left side IHC
Prim-5 to Long-pec lateral plate mesoderm right side IHC
WT + MO1-hs3st3b1b standard conditions Prim-5 to Long-pec lateral plate mesoderm left side IHC
WT + MO2-hs3st1l2 standard conditions Prim-5 to Long-pec Not Detected lateral plate mesoderm IHC
Prim-5 to Long-pec lateral plate mesoderm lateral side IHC
Prim-5 to Long-pec lateral plate mesoderm left side IHC
Prim-5 to Long-pec lateral plate mesoderm right side IHC
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-hs3st1l2 standard conditions Long-pec gut left/right pattern formation process quality, abnormal
Long-pec heart looping process quality, abnormal
Long-pec left/right pattern formation process quality, abnormal
Long-pec whole organism curved, abnormal
WT + MO2-hs3st1l2 standard conditions Long-pec gut left/right pattern formation process quality, abnormal
WT + MO2-hs3st1l2 standard conditions Long-pec heart looping process quality, abnormal
WT + MO2-hs3st1l2 standard conditions Long-pec left/right pattern formation process quality, abnormal
WT + MO2-hs3st1l2 standard conditions Long-pec whole organism curved, abnormal
twu34Tg + MO1-hs3st1l2 standard conditions Long-pec heart looping process quality, abnormal
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