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ZFIN ID: ZDB-FIG-140522-33
Ning et al., 2013 - MicroRNA-92a upholds Bmp signaling by targeting noggin3 during pharyngeal cartilage formation. Developmental Cell   24(3):283-295 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: 5-9 somites to Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage: Pec-fin

Fig. 2

Inactivation of mir92a Affects Late but Not Early Steps of Pharyngeal Cartilage Development

(A–E) The expression pattern of various markers in uninjected (UIC) or 92a-MO-injected (4 ng) embryos as detected by whole-mount in situ hybridization. Embryos in (A) and (B) were dorsal views with anterior to the top; the other embryos were lateral views with anterior to the left. The indicated ratio was the number of embryos with the shown representative phenotype/total number of observed embryos. Red and yellow circles in (D) indicated the pharyngeal region and pectoral fin, respectively. In (E), red and green circles indicated the pharyngeal arches and the cartilage capsule of the ears, and red arrowhead indicated the ethmoid plate and trabeculae cranii of dorsal neurocranial cartilages.

(F) The prechondrogenic condensation in mir92a morphants appeared normal. Embryos were injected at the one-cell stage with 4 ng 92a-rMO or 92a-MO, treated with BODIPY-ceramide at the end of gastrulation, and imaged by confocal microscopy at 48 hpf. The pharyngeal region was laterally viewed with dorsal to the top and anterior to the left. Extracellular space and cavity looked black. Arrows indicated aortic arches. The boxed area in the left image was enlarged and presented in the right. Scale bars, 20 µm.

(G) Detection of Col2 protein in pharyngeal prechondrogenic progenitors. Tg(fli1:EGFP) transgenic embryos were injected with 4 ng 92a-rMO or 92a-MO at the one-cell stage and coimmunostained with anti-GFP and anti-Col2 antibodies. The palatoquadrate (pq) and ceratohyal (ch) precartilages were shown with anterior to the top. Arrows in the right panel indicated eyes. Scale bars, 20 µm.

(H) Detection of Col2 in the neurocranial cartilages. Wild-type embryos, injected as in (G), were immunostained with anti-Col2 antibody. For each panel, the left was the dorsal view of the head region with anterior to the top and the right represented an enlargement of the area boxed in the left. Dorsal view, anterior to the top. The boxed area in the left image was presented at higher magnification in the corresponding right image. ch, ceratohyal; ep, ethmoid plate; pq, palatoquadrate; tc, trabeculate cranii. Scale bars, 50µm (left) and 5µm (right).

See also Fig. S2.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
col2a1a WT standard conditions Protruding-mouth auditory capsule ISH
Protruding-mouth chondrocyte ISH
Protruding-mouth ethmoid cartilage ISH
Protruding-mouth pharyngeal arch ISH
Protruding-mouth pharyngeal arch chondrocyte ISH
WT + MO2-mir92a standard conditions Protruding-mouth auditory capsule ISH
Protruding-mouth chondrocyte ISH
Protruding-mouth ethmoid cartilage ISH
Protruding-mouth Not Detected pharyngeal arch ISH
Protruding-mouth Not Detected pharyngeal arch chondrocyte ISH
dlx2a WT standard conditions Prim-5 ectomesenchyme ISH
WT + MO2-mir92a standard conditions Prim-5 ectomesenchyme ISH
EGFP y1Tg control Pec-fin ceratohyal cartilage IHC
Pec-fin palatoquadrate cartilage IHC
y1Tg + MO2-mir92a standard conditions Pec-fin ceratohyal cartilage IHC
Pec-fin palatoquadrate cartilage IHC
foxd3 WT standard conditions 5-9 somites cranial neural crest ISH
WT + MO2-mir92a standard conditions 5-9 somites cranial neural crest ISH
gsc WT standard conditions Long-pec auditory capsule ISH
Long-pec pharyngeal arch ISH
WT + MO2-mir92a standard conditions Long-pec auditory capsule ISH
Long-pec pharyngeal arch ISH
hand2 WT standard conditions Prim-25 ectomesenchyme ISH
WT + MO2-mir92a standard conditions Prim-25 ectomesenchyme ISH
sox9a WT standard conditions Prim-5 ectomesenchyme ISH
WT + MO2-mir92a standard conditions Prim-5 ectomesenchyme ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Qualifier Anatomy
Ab1-col2a IHC WT control Pec-fin ceratohyal cartilage
IHC Pec-fin ethmoid cartilage
IHC Pec-fin palatoquadrate cartilage
IHC Pec-fin trabecula cranii
IHC WT + MO2-mir92a standard conditions Pec-fin ethmoid cartilage
IHC Pec-fin trabecula cranii
IHC y1Tg control Pec-fin ceratohyal cartilage
IHC Pec-fin palatoquadrate cartilage
IHC y1Tg + MO2-mir92a standard conditions Pec-fin Not Detected ceratohyal cartilage
IHC Pec-fin Not Detected palatoquadrate cartilage
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO2-mir92a standard conditions Pec-fin chondrocranium chondrocyte differentiation process quality, abnormal
y1Tg + MO2-mir92a standard conditions Pec-fin ceratohyal cartilage chondroblast disorganized, abnormal
Pec-fin ceratohyal cartilage chondrocyte differentiation process quality, abnormal
Pec-fin chondrocranium chondrocyte differentiation process quality, abnormal
Pec-fin palatoquadrate cartilage chondroblast disorganized, abnormal
Pec-fin palatoquadrate cartilage chondrocyte differentiation process quality, abnormal
Pec-fin pharyngeal arch chondroblast disorganized, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Cell for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Cell, 24(3), Ning, G., Liu, X., Dai, M., Meng, A., and Wang, Q., MicroRNA-92a upholds Bmp signaling by targeting noggin3 during pharyngeal cartilage formation, 283-295, Copyright (2013) with permission from Elsevier. Full text @ Dev. Cell