PUBLICATION

Endodermal pouch-expressed dmrt2b is important for pharyngeal cartilage formation.

Authors: Li, L., Mao, A., Wang, P., Ning, G., Cao, Y., Wang, Q.
ID: ZDB-PUB-181021-1
Date: 2018
Source: Biology Open 7(12): (Journal)
Registered Authors: Wang, Qiang
Keywords: Craniofacial cartilage, Crossveinless 2, Cxcl12b, Dmrt2b, Endodermal pouch
MeSH Terms: none
PubMed: 30341107 Full text @ Biol. Open

Abstract

Pharyngeal pouches, a series of outpocketings derived from the foregut endoderm, are essential for craniofacial skeleton formation. However, the molecular mechanisms underlying endodermal pouch-regulated head cartilage development are not fully understood. In this study, we find that zebrafish dmrt2b, a gene encoding Doublesex and Mab-3-related transcription factor, is specifically expressed in endodermal pouches and required for normal pharyngeal cartilage development. Loss of dmrt2b doesn't affect cranial neural crest (CNC) specification and migration, but leads to prechondrogenic condensation defects by reducing cxcl12b expression after CNC cell movement into the pharyngeal arches. Moreover, dmrt2b inactivation results in reduced proliferation and impaired differentiation of CNC cells. We also show that dmrt2b suppresses crossveinless 2 expression in endodermal pouches to maintain BMP/Smad signaling in the arches, thereby facilitating CNC cell proliferation and chondrogenic differentiation. This work provides insight into how transcription factors expressed in endodermal pouches regulate pharyngeal skeleton development through tissue-tissue interactions.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
bmper	TU	control	Prim-25	pharyngeal pouch	ISH	Fig. 5
bmper	TU + MO3-dmrt2b	standard conditions	Prim-25	pharyngeal pouch	ISH	Fig. 5
col2a1a	dmrt2b^ioz21/ioz21 (TU)	standard conditions	Protruding-mouth	pharyngeal arch	ISH	Fig. 4
col2a1a	TU	control	Protruding-mouth	pharyngeal arch	ISH	Fig. 4
cxcl12b	dmrt2b^ioz21/ioz21 (TU)	standard conditions	Prim-5	pharyngeal arch cranial neural crest	ISH	Fig. 3
cxcl12b	TU	control	Prim-5	pharyngeal arch cranial neural crest	ISH	Fig. 3
cxcl12b	TU + MO3-dmrt2b	standard conditions	Prim-5	pharyngeal arch cranial neural crest	ISH	Fig. 3
dlx2a	dmrt2b^ioz21/ioz21 (TU)	standard conditions	Prim-5	migratory cranial neural crest	ISH	Fig. S4
dlx2a	dmrt2b^ioz21/ioz21 (TU)	standard conditions	14-19 somites	migratory cranial neural crest	ISH	Fig. S4
dlx2a	dmrt2b^ioz21/ioz21 (TU)	standard conditions	Prim-25	pharyngeal arch neural crest	ISH	Fig. 3

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
TU + MO2-sox32	standard conditions	75%-epiboly	endoderm sox17 expression absent, abnormal	Fig. 1
TU + MO2-sox32	standard conditions	Prim-25	pharyngeal pouch dmrt2b expression absent, abnormal	Fig. 1
TU + MO2-sox32	standard conditions	Prim-25	pharyngeal pouch nkx2.3 expression absent, abnormal	Fig. 1
TU + MO3-dmrt2b	standard conditions	Prim-5	pharyngeal arch cranial neural crest cxcl12b expression decreased amount, abnormal	Fig. 3
TU + MO3-dmrt2b	standard conditions	Prim-5	whole organism dmrt2b expression decreased amount, abnormal	Fig. S2
TU + MO3-dmrt2b	standard conditions	Prim-25	pharyngeal arch neural crest hand2 expression decreased amount, abnormal	Fig. 3
TU + MO3-dmrt2b	standard conditions	Prim-25	pharyngeal pouch bmper expression increased amount, abnormal	Fig. 5
TU + MO3-dmrt2b	standard conditions	Day 4	auditory capsule morphology, abnormal	Fig. 2
TU + MO3-dmrt2b	standard conditions	Day 4	ceratobranchial 1 cartilage decreased size, abnormal	Fig. 2
TU + MO3-dmrt2b	standard conditions	Day 4	ceratobranchial 1 cartilage morphology, abnormal	Fig. 2

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
ba2Tg	Tg(-4.9sox10:EGFP)	Transgenic Insertion
ioz15Tg	Tg(-5.5nkx2.3:mCherry)	Transgenic Insertion
ioz19Tg	Tg(-5.5nkx2.3:KALTA4-mCherry)	Transgenic Insertion
ioz21		Small Deletion	dmrt2b
ioz22Tg	Tg(-4.9sox10:mCherry-CAAX)	Transgenic Insertion
pt510Tg	Tg1(BMPRE:EGFP)	Transgenic Insertion
s870Tg	Tg(sox17:GFP)	Transgenic Insertion
y1Tg	Tg(fli1:EGFP)	Transgenic Insertion

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
bmper	MO1-bmper	MRPHLNO
dmrt2b	CRISPR1-dmrt2b	CRISPR
dmrt2b	MO3-dmrt2b	MRPHLNO
sox32	MO2-sox32	MRPHLNO

Fish

Fish
ba2Tg
ba2Tg; ioz15Tg
ba2Tg; ioz15Tg + MO3-dmrt2b
ba2Tg + MO3-dmrt2b
dmrt2b^ioz21/ioz21 + MO1-bmper (TU)
dmrt2b^ioz21/ioz21 (TU)
ioz19Tg
ioz19Tg; s870Tg
ioz22Tg
ioz22Tg + MO3-dmrt2b

Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab1-col2a	monoclonal		IgG1	Mouse
Ab1-smad	polyclonal	smad1 smad5 smad9	IgG	Rabbit
Ab36-h3	monoclonal		IgG	Rabbit

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP
GFP	EFG	GFP
KALTA4	EFG	KALTA4
mCherry	EFG	mCherry

Mapping

Li et al., 2018 ZDB-PUB-181021-1 PMID:30341107

Endodermal pouch-expressed dmrt2b is important for pharyngeal cartilage formation.

Li et al., 2018

ZDB-PUB-181021-1
PMID:30341107