header logo image header logo text
Downloads Login
Research
General Information
ZIRC
ZFIN ID: ZDB-FIG-150113-16
Muto et al., 2014 - Nipbl and Mediator Cooperatively Regulate Gene Expression to Control Limb Development. PLoS Genetics   10:e1004671 Full text @ PLoS Genet.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Prim-25 to Long-pec
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Long-pec to Day 5

Fig. 7 Med12 depletion disrupts pectoral fin morphology and gene expression similar to Nipbl depletion.

(A, B) Morphology of live embryos at 52 hpf (A, lateral views) and 76 hpf (B, dorsal views). (A) Anterior halves of control and med12-MO-injected embryos (left column) and higher magnification pictures of their pectoral fin buds (right column). (B) Dorsal views of embryos at 76 hpf. (C) Whisker plots of fin length. Fin lengths (medians) are 430.0 μm, n = 18 (control), 275.6 μm, n = 20 (med12-MO, 2 ng), and 183.8 μm, n = 20 (med12-MO, 4 ng). *: p<10-4. (D) Alcian blue staining of pectoral fin cartilage of control (upper) and Med12-deficient (med12-MO, 4 ng; lower) embryos at 120 hpf. Dorsal view, anterior to the top. Right column, higher magnification pictures of boxed areas of endoskeletal discs. ac, actinotrichs; ed, endoskeletal disc; sco, scapulocoracoid. (E) Controls for med12-MO efficiency. RT-PCR, 36 hpf. Both pairs of med12 primers (Primer #1 and #2) show that splicing of med12 mRNA is significantly suppressed by med12-MO, with a slightly higher efficiency at 6 ng. Primer pair #1 detects both precursor and mature mRNA, whereas primer pair #2 only detects mature mRNA (see Materials and Methods). nipbla and nipblb expression was not affected by Med12 depletion. ef1a was used as a control. (F) Expression of genes involved in the 5′-hox/hand2/shha gene cassette and AER fgf genes in pectoral fin buds examined by ISH at 36 hpf. Dorsal views, anterior to the top. Similar to Nipbl-deficient embryos, shha, hand2 and 5′-hoxd genes in mesenchyme as well as fgf16 and fgf8a in the AER are reduced in Med12-deficient embryos (4 ng/embryo med12-MO).

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
fgf8a AB control Long-pec fin fold pectoral fin bud ISH
AB + MO5-med12 standard conditions Long-pec Not Detected fin fold pectoral fin bud ISH
fgf16 AB control Long-pec fin fold pectoral fin bud ISH
AB + MO5-med12 standard conditions Long-pec Not Detected fin fold pectoral fin bud ISH
hand2 AB control Long-pec pectoral fin bud ISH
AB + MO5-med12 standard conditions Long-pec pectoral fin bud ISH
hoxd9a AB control Long-pec pectoral fin bud ISH
AB + MO5-med12 standard conditions Long-pec pectoral fin bud ISH
hoxd10a AB control Long-pec pectoral fin bud ISH
AB + MO5-med12 standard conditions Long-pec pectoral fin bud ISH
hoxd11a AB control Long-pec pectoral fin bud ISH
AB + MO5-med12 standard conditions Long-pec pectoral fin bud ISH
hoxd13a AB control Long-pec pectoral fin bud ISH
AB + MO5-med12 standard conditions Long-pec Not Detected pectoral fin bud ISH
nipbla WT control Prim-25 whole organism RTPCR
WT + MO5-med12 standard conditions Prim-25 whole organism RTPCR
nipblb WT control Prim-25 whole organism RTPCR
WT + MO5-med12 standard conditions Prim-25 whole organism RTPCR
shha AB control Long-pec pectoral fin bud ISH
AB + MO5-med12 standard conditions Long-pec pectoral fin bud ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
AB + MO5-med12 standard conditions Long-pec pectoral fin bud embryonic pectoral fin morphogenesis decreased process quality, abnormal
Protruding-mouth pectoral fin decreased length, abnormal
Day 5 pectoral fin cartilage malformed, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS Genetics for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS Genet.