ZFIN ID: ZDB-FIG-180608-104
Barske et al., 2018 - Essential Role of Nr2f Nuclear Receptors in Patterning the Vertebrate Upper Jaw. Developmental Cell   44(3):337-347.e5 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Anatomical Terms:
Stage Range: 1-4 somites to Day 6
PHENOTYPE:
Fish:
Observed In:
Stage Range: 1-4 somites to Day 6

Fig. 2

Transformation of the Upper Jaw in Nr2f Mutants

(A–C) Larval heads stained for cartilage (Alcian blue) and bone (Alizarin red). Boxed area in (A) represents the approximate region of the dissected upper- and lower-jaw cartilages in A′–C′. In mutants (B′–C′), the pterygoid process (Ptp, green in A″) of the palatoquadrate is thickened (black arrows) to resemble Meckel's (magenta in A″). Skeletal structures derived from the dorsal hyoid arch are reduced in the triple mutant (magenta arrow in C).

(D) Frequency of the thickened Ptp phenotype across different nr2f1b; nr2f2; nr2f5 genotypes.

(E) Some nr2f2; nr2f5 mutants form an ectopic process (ra) that resembles the retroarticular process (ra) of Meckel's.

(F) Fluorescent in situ images show expansion of sox9a expression into the mutant maxillary prominence (white arrow; dlx2a labels all arch NCCs).

(G) Formation of a larger cartilage (sox10:DsRed+, magenta) in the mutant is accompanied by a reduction in the number of Sp7+ osteoblasts (green) in the neighboring dermal entopterygoid bone (en; white double arrowhead). Mutants have normal numbers of Sp7+ osteoblasts in the lower-jaw dentary bone (den).

(H) sox10+ NCCs are specified at the neural plate border in all Nr2f mutants (n = 18 nr2f1aany; nr2f1bany; nr2f2−/−; nr2f5−/−; n = 2 quadruple). The nr2f2; nr2f5 double mutant shown is also nr2f1a+−-; nr2f1b−/−.

(I) Three streams of Sox10+ NCCs (white arrows) are evident in all mutant combinations (n = 13 nr2f1aany; nr2f1bany; nr2f2−/−; nr2f5−/−) except the quadruple mutant (n = 1). Sox10+ otic cells (yellow double arrow) are indicated for reference.

Scale bars: (C and E) 100 μm; (F–I) 50 μm. Also see Figure S2 and Table S4 for mutation details and skeletal phenotypes of single and other combinatorial mutants.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
dlx2a TU standard conditions Prim-25 pharyngeal arch 1 neural crest cell ISH
Prim-25 pharyngeal arch 2 neural crest cell ISH
nr2f1bel523/+; nr2f2el506/el506; nr2f5el611/el611 (TU) standard conditions Prim-25 pharyngeal arch 1 neural crest cell ISH
Prim-25 pharyngeal arch 2 neural crest cell ISH
DsRed el10Tg (TU) standard conditions Day 6 chondrocyte IHC
nr2f1bel523/+; nr2f2el506/el506; nr2f5el611/el611; el10Tg (TU) standard conditions Day 6 chondrocyte IHC
sox9a TU standard conditions Prim-25 maxilla ISH
Prim-25 Not Detected maxilla bony projection ISH
nr2f1bel523/+; nr2f2el506/el506; nr2f5el611/el611 (TU) standard conditions Prim-25 maxilla ISH
Prim-25 maxilla bony projection ISH
sox10 TU standard conditions 1-4 somites neural crest ISH
nr2f1ael512/el512; nr2f1bel523/el523; nr2f2el506/el506; nr2f5el611/el611 (TU) standard conditions 1-4 somites neural crest ISH
nr2f2el506/el506; nr2f5el611/el611 (TU) standard conditions 1-4 somites neural crest ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-sp7 IHC el10Tg (TU) standard conditions Day 6 dentary osteoblast
IHC Day 6 entopterygoid osteoblast
IHC nr2f1bel523/+; nr2f2el506/el506; nr2f5el611/el611; el10Tg (TU) standard conditions Day 6 dentary osteoblast
IHC Day 6 entopterygoid osteoblast
Ab3-sox10 IHC TU standard conditions 14-19 somites neural crest
IHC nr2f1ael512/el512; nr2f1bel523/el523; nr2f2el506/el506; nr2f5el611/el611 (TU) standard conditions 14-19 somites neural crest
IHC nr2f2el506/el506; nr2f5el611/el611 (TU) standard conditions 14-19 somites neural crest
Phenotype Details
Fish Conditions Stage Phenotype
nr2f1ael512/el512; nr2f1bel523/el523; nr2f2el506/el506; nr2f5el611/el611 (TU) standard conditions 1-4 somites neural crest cell migration decreased occurrence, abnormal
nr2f1bel523/+; nr2f2el506/el506; nr2f5el611/el611 (TU) standard conditions Prim-25 maxilla sox9a expression increased distribution, abnormal
Prim-25 maxilla bony projection sox9a expression increased amount, abnormal
Prim-25 maxilla bony projection sox9a expression mislocalised, abnormal
Day 5 palatoquadrate arch increased size, abnormal
Day 5 pterygoid process increased thickness, abnormal
Day 5 retroarticular increased amount, abnormal
nr2f1bel523/+; nr2f2el506/el506; nr2f5el611/el611; el10Tg (TU) standard conditions Day 6 entopterygoid osteoblast decreased amount, abnormal
nr2f1bel523/el523; nr2f2el506/el506; nr2f5el611/el611 (TU) standard conditions Day 5 dorsal hyoid arch decreased size, abnormal
Day 5 palatoquadrate arch increased size, exacerbated
Day 5 pterygoid process increased thickness, abnormal
nr2f2el506/el506; nr2f5el611/el611 (TU) standard conditions Day 5 palatoquadrate arch increased size, abnormal
Acknowledgments:
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Reprinted from Developmental Cell, 44(3), Barske, L., Rataud, P., Behizad, K., Del Rio, L., Cox, S.G., Crump, J.G., Essential Role of Nr2f Nuclear Receptors in Patterning the Vertebrate Upper Jaw, 337-347.e5, Copyright (2018) with permission from Elsevier. Full text @ Dev. Cell