PUBLICATION

Failed Progenitor Specification Underlies the Cardiopharyngeal Phenotypes in a Zebrafish Model of 22q11.2 Deletion Syndrome

Authors

Guner-Ataman, B., González-Rosa, J.M., Shah, H.N., Butty, V.L., Jeffrey, S., Abrial, M., Boyer, L.A., Burns, C.G., Burns, C.E.

ID

ZDB-PUB-180802-10

Date

2018

Source

Cell Reports 24: 1342-1354.e5 (Journal)

Registered Authors

Burns (Erter), Caroline, Burns, Geoff, Gonzalez-Rosa, Juan Manuel

Keywords

22q11, DiGeorge, Tbx1, arch artery, cardiopharyngeal, heart, nkx2.5, progenitor, zebrafish

Datasets

GEO:GSE103120

MeSH Terms

Homeobox Protein Nkx-2.5/genetics
Homeobox Protein Nkx-2.5/metabolism
Cell Differentiation*
Cell Lineage
Zebrafish
Pharynx/cytology
Pharynx/embryology*
T-Box Domain Proteins/genetics
22q11 Deletion Syndrome/genetics
22q11 Deletion Syndrome/pathology*
Phenotype
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism
Animals
Embryonic Stem Cells/cytology*
Embryonic Stem Cells/metabolism

(all 16)

PubMed

30067987 Full text @ Cell Rep.

Abstract

Microdeletions involving TBX1 result in variable congenital malformations known collectively as 22q11.2 deletion syndrome (22q11.2DS). Tbx1-deficient mice and zebrafish recapitulate several disease phenotypes, including pharyngeal arch artery (PAA), head muscle (HM), and cardiac outflow tract (OFT) deficiencies. In zebrafish, these structures arise from nkx2.5⁺ progenitors in pharyngeal arches 2-6. Because pharyngeal arch morphogenesis is compromised in Tbx1-deficient animals, the malformations were considered secondary. Here, we report that the PAA, HM, and OFT phenotypes in tbx1 mutant zebrafish are primary and arise prior to pharyngeal arch morphogenesis from failed specification of the nkx2.5⁺ pharyngeal lineage. Through in situ analysis and lineage tracing, we reveal that nkx2.5 and tbx1 are co-expressed in this progenitor population. Furthermore, we present evidence suggesting that gdf3-ALK4 signaling is a downstream mediator of nkx2.5⁺ pharyngeal lineage specification. Collectively, these studies support a cellular mechanism potentially underlying the cardiovascular and craniofacial defects observed in the 22q11.2DS population.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
AmCyan	fb23Tg; fb3Tg	standard conditions	Day 4	lateral dorsal aorta	IFL	Fig. 4
AmCyan	fb23Tg; fb3Tg	standard conditions	Day 4	pharyngeal arch 6	IFL	Fig. 4
gdf3	tbx1^tu285/tu285	standard conditions	5-9 somites	anterior lateral plate mesoderm	ISH	Fig. 5
gdf3	WT	control	5-9 somites	anterior lateral plate mesoderm	ISH	Fig. 5
GFP	cn2Tg; fb23Tg	standard conditions	Protruding-mouth	adductor mandibulae	IHC	Fig. 4
GFP	cn2Tg; fb23Tg	standard conditions	Protruding-mouth	bulbus arteriosus cardiac muscle cell	IHC	Fig. 4
GFP	cn2Tg; fb23Tg	standard conditions	Protruding-mouth	bulbus arteriosus endothelial cell	IHC	Fig. 4
GFP	cn2Tg; fb23Tg	standard conditions	Protruding-mouth	bulbus arteriosus smooth muscle	IHC	Fig. 4
GFP	cn2Tg; fb23Tg	standard conditions	Protruding-mouth	cardiac ventricle	IHC	Fig. 4
GFP	cn2Tg; fb23Tg	standard conditions	Protruding-mouth	cranial vasculature	IHC	Fig. S3

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	chemical treatment by environment: SB 505124	5-9 somites	anterior lateral plate mesoderm nkx2.5 expression decreased amount, abnormal	Fig. 7
WT	chemical treatment by environment: SB 505124	5-9 somites	anterior lateral plate mesoderm nkx2.5 expression decreased distribution, abnormal	Fig. 7
WT	chemical treatment by environment: SB 505124	Prim-5	whole organism morphology, abnormal	Fig. S7
WT	chemical treatment by environment: SB 505124	Prim-25	pharyngeal arch 3 tie1 expression decreased amount, abnormal	Fig. 6
WT	chemical treatment by environment: SB 505124	Prim-25	pharyngeal arch 3 tie1 expression decreased distribution, abnormal	Fig. 6
WT	chemical treatment by environment: SB 505124	Prim-25	pharyngeal arch 4 tie1 expression decreased amount, abnormal	Fig. 6
WT	chemical treatment by environment: SB 505124	Prim-25	pharyngeal arch 4 tie1 expression decreased distribution, abnormal	Fig. 6
WT	chemical treatment by environment: SB 505124	Prim-25	pharyngeal arch 5 tie1 expression absent, abnormal	Fig. 6
WT	chemical treatment by environment: SB 505124	Protruding-mouth	bulbus arteriosus smooth muscle ab1-elnb labeling decreased amount, abnormal	Fig. 6
WT	chemical treatment by environment: SB 505124	Protruding-mouth	bulbus arteriosus smooth muscle ab1-elnb labeling decreased distribution, abnormal	Fig. 6

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
cn2Tg	Tg(-3.5ubb:LOXP-LacZ-LOXP-egfp)	Transgenic Insertion
fb3Tg	TgBAC(kdrl:LOXP-AmCyan-LOXP-ZsYellow)	Transgenic Insertion
fb7Tg	TgBAC(-36nkx2.5:ZsYellow)	Transgenic Insertion
fb9Tg	TgBAC(nkx2.5:Kaede)	Transgenic Insertion
fb20a		Small Deletion	gdf3
fb23Tg	TgBAC(tbx1:mKate2-2A-Cre)	Transgenic Insertion
fb25Tg	TgBAC(tbx1:GFP)	Transgenic Insertion
la116Tg	Tg(kdrl:GFP)	Transgenic Insertion
tu285		Point Mutation	tbx1

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Human Disease / Model

Human Disease	Fish	Conditions	Evidence
velocardiofacial syndrome			TAS

Sequence Targeting Reagents

Target	Reagent	Reagent Type
gdf3	CRISPR3-gdf3	CRISPR

Fish

Fish
cn2Tg; fb23Tg
fb7Tg
fb9Tg
fb23Tg
fb23Tg; fb3Tg
fb25Tg
gdf3^fb20a/fb20a
gdf3^fb20a/fb20a
la116Tg
tbx1^tu285/tu285

Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab1-elnb	polyclonal	elnb	IgG	Rabbit
Ab-MF20	monoclonal		IgG2b	Mouse

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
AmCyan	EFG	AmCyan
Cre	EFG	Cre
GFP	EFG	GFP
Kaede	EFG	Kaede
LacZ	EFG	LacZ
mKate2	EFG	mKate2
ZsYellow	EFG	ZsYellow

Mapping

Guner-Ataman et al., 2018 ZDB-PUB-180802-10 PMID:30067987

Failed Progenitor Specification Underlies the Cardiopharyngeal Phenotypes in a Zebrafish Model of 22q11.2 Deletion Syndrome

Guner-Ataman et al., 2018

ZDB-PUB-180802-10
PMID:30067987