PUBLICATION

The knockdown of maternal glucocorticoid receptor mRNA alters embryo development in zebrafish

Authors

Pikulkaew, S., Benato, F., Celeghin, A., Zucal, C., Skobo, T., Colombo, L., and Dalla Valle , L.

ID

ZDB-PUB-110317-12

Date

2011

Source

Developmental Dynamics : an official publication of the American Association of Anatomists 240(4): 874-889 (Journal)

Registered Authors

Benato, Francesca, Dalla Valle, Luisa

Keywords

zebrafish, gr mRNA, morpholino (MO), development

Datasets

GEO:GSE25517

MeSH Terms

Animals
Base Sequence
Animals, Genetically Modified
Zebrafish/embryology*
Zebrafish/genetics*
Zebrafish/metabolism
Larva/genetics
Larva/growth & development
Larva/metabolism
Embryo, Nonmammalian
Gene Expression Regulation, Developmental
Female
Apoptosis/genetics
Apoptosis/physiology
RNA, Messenger/genetics
RNA, Messenger/metabolism
Gene Knockdown Techniques
Receptors, Glucocorticoid/genetics*
Receptors, Glucocorticoid/metabolism
Molecular Sequence Data
RNA, Messenger, Stored/genetics*
RNA, Messenger, Stored/metabolism
Embryonic Development/genetics*

(all 23)

PubMed

21360790 Full text @ Dev. Dyn.

Abstract

In zebrafish, ovulated oocytes contain both maternal cortisol and the mRNA for the glucocorticoid receptor (gr), which is spread as granular structures throughout the ooplasm. At 0.2 hpf, this transcript is relocated in the blastodisc area and partitioned among blastomeres. At 6-8 hpf, it is replaced by zygotic transcript. We used morpholinos to block translation of both maternal and zygotic gr transcripts, and a missplicing morpholino to block post-transcriptionally the zygotic transcript alone. Only knockdown of translation produced an increase of apoptosis and subsequent craniofacial and caudal deformities with severe malformations of neural, vascular, and visceral organs in embryos and 5-dpf larvae. Such defects were rescued with trout gr2 mRNA. Microarray analysis revealed that 114 and 37 highly expressed transcripts were up- and down-regulated, respectively, by maternal Gr protein deficiency in 5-hpf embryos. These results indicate that the maternal gr transcript and protein participate in the maternal programming of zebrafish development.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
casp8	WT	standard conditions	Sphere	whole organism	RTPCR	Fig. 7
casp8	WT	standard conditions	75%-epiboly	whole organism	RTPCR	Fig. 7
casp8	WT	standard conditions	5-9 somites	whole organism	RTPCR	Fig. 7
casp8	WT + MO2-nr3c1	standard conditions	75%-epiboly	whole organism	RTPCR	Fig. 7
casp8	WT + MO2-nr3c1	standard conditions	5-9 somites	whole organism	RTPCR	Fig. 7
casp8	WT + MO2-nr3c1	standard conditions	Sphere	whole organism	RTPCR	Fig. 7
chrd	WT	control	Bud	axial hypoblast	ISH	Fig. s1
chrd	WT	control	Shield	shield	ISH	Fig. s1
chrd	WT + MO2-nr3c1	standard conditions	Bud	axial hypoblast	ISH	Fig. s1
chrd	WT + MO2-nr3c1	standard conditions	Shield	shield	ISH	Fig. s1

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT + MO2-nr3c1	standard conditions	Day 6	ceratohyal cartilage disoriented, abnormal	Fig. 5
WT + MO2-nr3c1	standard conditions	Day 6	ceratohyal cartilage fused with hypobranchial cartilage, abnormal	Fig. 5
WT + MO2-nr3c1	standard conditions	Day 6	ethmoid cartilage curled, abnormal	Fig. 5
WT + MO2-nr3c1	standard conditions	Day 6	ethmoid cartilage retracted, abnormal	Fig. 5
WT + MO2-nr3c1	standard conditions	Day 6	Meckel's cartilage decreased size, abnormal	Fig. 5
WT + MO2-nr3c1	standard conditions	Day 6	Meckel's cartilage disoriented, abnormal	Fig. 5
WT + MO2-nr3c1	standard conditions	Day 6	Meckel's cartilage fused with palatoquadrate cartilage, abnormal	Fig. 5
WT + MO2-nr3c1	standard conditions	Prim-5	apoptotic process increased occurrence, abnormal	Fig. 4
WT + MO2-nr3c1	standard conditions	Prim-5	embryo development delayed, abnormal	Fig. 3
WT + MO2-nr3c1	standard conditions	Prim-5	eye decreased size, abnormal	Fig. 3

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
y1Tg	Tg(fli1:EGFP)	Transgenic Insertion

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
nr3c1	MO2-nr3c1	MRPHLNO
nr3c1	MO3-nr3c1	MRPHLNO
nr3c1	MO4-nr3c1	MRPHLNO
tp53	MO4-tp53	MRPHLNO

Fish

Fish
WT
WT + MO2-nr3c1
WT + MO3-nr3c1
WT + MO4-nr3c1
y1Tg
y1Tg + MO2-nr3c1

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP

Mapping

Pikulkaew et al., 2011 ZDB-PUB-110317-12 PMID:21360790

The knockdown of maternal glucocorticoid receptor mRNA alters embryo development in zebrafish

Pikulkaew et al., 2011

ZDB-PUB-110317-12
PMID:21360790