PUBLICATION

Embryonic Ethanol Exposure Dysregulates BMP and Notch Signaling, Leading to Persistent Atrio-Ventricular Valve Defects in Zebrafish

Authors

Sarmah, S., Muralidharan, P., Marrs, J.A.

ID

ZDB-PUB-160825-2

Date

2016

Source

PLoS One 11: e0161205 (Journal)

Registered Authors

Marrs, James A., Sarmah, Swapnalee

Keywords

Embryos, Ethanol, Heart, Zebrafish, Cardiac ventricles, Endocardium, BMP signaling, Larvae

MeSH Terms

Heart Valves/abnormalities
Heart Valves/embryology
Heart Valves/metabolism
Endocardium/drug effects
Endocardium/embryology
Endocardium/metabolism
Embryonic Development/drug effects*
Embryonic Development/genetics
Animals
Ethanol/adverse effects*
Receptors, Notch/genetics
Receptors, Notch/metabolism*
Zebrafish
Embryo, Nonmammalian*/drug effects
Embryo, Nonmammalian*/metabolism
Signal Transduction/drug effects*
Gene Expression Regulation, Developmental
Bone Morphogenetic Proteins/genetics
Bone Morphogenetic Proteins/metabolism*
Organogenesis/drug effects
Heart Defects, Congenital/etiology*
Heart Defects, Congenital/pathology
Heart Ventricles/abnormalities
Heart Ventricles/embryology
Heart Ventricles/metabolism
Biomarkers
Wnt Proteins/metabolism

(all 27)

PubMed

27556898 Full text @ PLoS One

Abstract

Fetal alcohol spectrum disorder (FASD), birth defects associated with ethanol exposure in utero, includes a wide spectrum of congenital heart defects (CHDs), the most prevalent of which are septal and conotruncal defects. Zebrafish FASD model was used to dissect the mechanisms underlying FASD-associated CHDs. Embryonic ethanol exposure (3-24 hours post fertilization) led to defects in atrio-ventricular (AV) valvulogenesis beginning around 37 hpf, a morphogenetic event that arises long after ethanol withdrawal. Valve leaflets of the control embryos comprised two layers of cells confined at the compact atrio-ventricular canal (AVC). Ethanol treated embryos had extended AVC and valve forming cells were found either as rows of cells spanning the AVC or as unorganized clusters near the AV boundary. Ethanol exposure reduced valve precursors at the AVC, but some ventricular cells in ethanol treated embryos exhibited few characteristics of valve precursors. Late staged larvae and juvenile fish exposed to ethanol during embryonic development had faulty AV valves. Examination of AVC morphogenesis regulatory networks revealed that early ethanol exposure disrupted the Bmp signaling gradient in the heart during valve formation. Bmp signaling was prominent at the AVC in controls, but ethanol-exposed embryos displayed active Bmp signaling throughout the ventricle. Ethanol exposure also led to mislocalization of Notch signaling cells in endocardium during AV valve formation. Normally, highly active Notch signaling cells were organized at the AVC. In ethanol-exposed embryos, highly active Notch signaling cells were dispersed throughout the ventricle. At later stages, ethanol-exposed embryos exhibited reduced Wnt/β-catenin activity at the AVC. We conclude that early embryonic ethanol exposure alters Bmp, Notch and other signaling activities during AVC differentiation leading to faulty valve morphogenesis and valve defects persist in juvenile fish.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
bmp4	WT	control	Long-pec	atrioventricular canal	ISH	Fig. 5
bmp4	WT	chemical treatment by environment: ethanol	Long-pec	atrioventricular canal	ISH	Fig. 5
bmp4	WT	chemical treatment by environment: ethanol	Long-pec	bulbus arteriosus	ISH	Fig. 5
bmp4	WT	control	Long-pec	bulbus arteriosus	ISH	Fig. 5
bmp4	WT	chemical treatment by environment: ethanol	Long-pec	cardiac ventricle	ISH	Fig. 5
d2EGFP	hsc6Tg; kyu1Tg	control	Long-pec	atrioventricular canal cardiac muscle cell	IFL	Fig. 9
d2EGFP	hsc6Tg; kyu1Tg	chemical treatment by environment: ethanol	Long-pec	atrioventricular canal cardiac muscle cell	IFL	Fig. 9
d2EGFP	hsc6Tg; kyu1Tg	chemical treatment by environment: ethanol	Protruding-mouth	atrioventricular canal cardiac muscle cell	IFL	Fig. 9
d2EGFP	hsc6Tg; kyu1Tg	control	Protruding-mouth	atrioventricular canal cardiac muscle cell	IFL	Fig. 9
d2EGFP	hsc6Tg; kyu1Tg	control	Protruding-mouth	cardiac ventricle cardiac muscle cell	IFL	Fig. 9

1 - 10 of 112

Show

Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	chemical treatment by environment: ethanol	Long-pec	atrioventricular valve cell decreased amount, abnormal	Fig. 2
WT	chemical treatment by environment: ethanol	Long-pec	atrioventricular valve cell displaced to cardiac ventricle, abnormal	Fig. 2
WT	chemical treatment by environment: ethanol	Long-pec	cardiac jelly mislocalised, abnormal	Fig. 5
WT	chemical treatment by environment: ethanol	Long-pec	cardiac ventricle bmp4 expression mislocalised, abnormal	Fig. 5
WT	chemical treatment by environment: ethanol	Long-pec	cardiac ventricle ab1-smad labeling mislocalised, abnormal	Fig. 5
WT	chemical treatment by environment: ethanol	Long-pec	cardiac ventricle endocardium ab1-smad labeling mislocalised, abnormal	Fig. 5
WT	chemical treatment by environment: ethanol	Long-pec	cardiac ventricle endocardium zn-5 labeling mislocalised, abnormal	Fig. 2
WT	chemical treatment by environment: ethanol	Long-pec	ventricular endocardium ab1-smad labeling mislocalised, abnormal	Fig. 5
WT	chemical treatment by environment: ethanol	Long-pec	ventricular endocardium zn-5 labeling mislocalised, abnormal	Fig. 2
WT	chemical treatment by environment: ethanol	Long-pec	ventricular myocardium ab1-smad labeling mislocalised, abnormal	Fig. 5

1 - 10 of 74

Show

Mutations / Transgenics

Allele	Construct	Type
hsc6Tg	Tg(myl7:NLS-KikGR)	Transgenic Insertion
jh11Tg	Tg(EPV.Tp1-Ocu.Hbb2:hmgb1-mCherry)	Transgenic Insertion
kyu1Tg	Tg(OTM:d2EGFP)	Transgenic Insertion
la116Tg	Tg(kdrl:GFP)	Transgenic Insertion
pt510Tg	Tg1(BMPRE:EGFP)	Transgenic Insertion
twu34Tg	Tg(myl7:EGFP)	Transgenic Insertion
y1Tg	Tg(fli1:EGFP)	Transgenic Insertion

1 - 7 of 7

Show

Human Disease / Model

Human Disease	Fish	Conditions	Evidence
fetal alcohol spectrum disorder	WT	chemical treatment by environment: ethanol	TAS

Sequence Targeting Reagents

Fish

Fish
hsc6Tg; kyu1Tg
jh11Tg
jh11Tg; la116Tg
jh11Tg; twu34Tg
la116Tg
pt510Tg
WT
y1Tg

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
d2EGFP	EFG	d2EGFP
EGFP	EFG	EGFP
GFP	EFG	GFP
KikGR	EFG	KikGR
mCherry	EFG	mCherry

Mapping

Sarmah et al., 2016 ZDB-PUB-160825-2 PMID:27556898

Embryonic Ethanol Exposure Dysregulates BMP and Notch Signaling, Leading to Persistent Atrio-Ventricular Valve Defects in Zebrafish

Sarmah et al., 2016

ZDB-PUB-160825-2
PMID:27556898