PUBLICATION

Actomyosin regulation by Eph receptor signaling couples boundary cell formation to border sharpness

Authors

Cayuso, J., Xu, Q., Addison, M., Wilkinson, D.G.

ID

ZDB-PUB-190911-8

Date

2019

Source

eLIFE 8: (Journal)

Registered Authors

Wilkinson, David, Xu, Qiling

Keywords

Eph receptor, actomyosin, border sharpening, cell specification, developmental biology, hindbrain, tension, zebrafish

MeSH Terms

Protein Processing, Post-Translational
Zebrafish Proteins/metabolism
Nuclear Proteins/metabolism
Gene Expression Regulation, Developmental*
Zebrafish
Brain/embryology*
DNA-Binding Proteins/metabolism
Transcription Factors/metabolism
Animals
Receptor, EphA4/metabolism*
Signal Transduction*
Actomyosin/metabolism*
Phosphorylation

(all 13)

PubMed

31502954 Full text @ Elife

Abstract

The segregation of cells with distinct regional identity underlies formation of a sharp border, which in some tissues serves to organise a boundary signaling centre. It is unclear whether or how border sharpness is coordinated with induction of boundary-specific gene expression. We show that forward signaling of EphA4 is required for border sharpening and induction of boundary cells in the zebrafish hindbrain, which we find both require kinase-dependent signaling, with a lesser input of PDZ domain-dependent signaling. We find that boundary-specific gene expression is regulated by myosin II phosphorylation, which increases actomyosin contraction downstream of EphA4 signaling. Myosin phosphorylation leads to nuclear translocation of Taz, which together with Tead1a is required for boundary marker expression. Since actomyosin contraction maintains sharp borders, there is direct coupling of border sharpness to boundary cell induction that ensures correct organisation of signaling centres.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
egr2b	efnb3b^fci9/fci9	standard conditions	14-19 somites	rhombomere 3	ISH	Figure 2
egr2b	efnb3b^fci9/fci9	standard conditions	14-19 somites	rhombomere 5	ISH	Figure 2
egr2b	epha4a^fci5/fci5	standard conditions	14-19 somites	rhombomere 3	ISH	Figure 2
egr2b	epha4a^fci5/fci5	standard conditions	14-19 somites	rhombomere 5	ISH	Figure 2
egr2b	epha4a^fci6/fci6	standard conditions	14-19 somites	rhombomere 3	ISH	Figure 2
egr2b	epha4a^fci6/fci6	standard conditions	14-19 somites	rhombomere 5	ISH	Figure 2
egr2b	epha4a^fci7/fci7	standard conditions	14-19 somites	rhombomere 3	ISH	Figure 2
egr2b	epha4a^fci7/fci7	standard conditions	14-19 somites	rhombomere 5	ISH	Figure 2
egr2b	epha4a^fci8/fci8	standard conditions	14-19 somites	rhombomere 3	ISH	Figure 2
egr2b	epha4a^fci8/fci8	standard conditions	14-19 somites	rhombomere 5	ISH	Figure 2

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	chemical treatment: blebbistatin	20-25 somites	rhombomere 1 posterior margin rfng expression decreased distribution, abnormal	Figure 3
WT	chemical treatment: blebbistatin	20-25 somites	rhombomere 2 posterior margin rfng expression decreased distribution, abnormal	Figure 3
WT	chemical treatment: blebbistatin	20-25 somites	rhombomere 3 posterior margin rfng expression decreased distribution, abnormal	Figure 3
WT	chemical treatment: blebbistatin	20-25 somites	rhombomere 4 posterior margin rfng expression decreased distribution, abnormal	Figure 3
WT	chemical treatment: blebbistatin	20-25 somites	rhombomere 5 posterior margin rfng expression decreased distribution, abnormal	Figure 3
WT	chemical treatment: blebbistatin	20-25 somites	rhombomere 6 posterior margin rfng expression decreased distribution, abnormal	Figure 3
WT + CRISPR1-ppp1r12a	standard conditions	20-25 somites	rhombomere 1 posterior margin rfng expression increased distribution, abnormal	Figure 3
WT + CRISPR1-ppp1r12a	standard conditions	20-25 somites	rhombomere 2 posterior margin rfng expression increased distribution, abnormal	Figure 3
WT + CRISPR1-ppp1r12a	standard conditions	20-25 somites	rhombomere 3 posterior margin rfng expression increased distribution, abnormal	Figure 3
WT + CRISPR1-ppp1r12a	standard conditions	20-25 somites	rhombomere 4 posterior margin rfng expression increased distribution, abnormal	Figure 3

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Mutations / Transgenics

Allele	Type	Affected Genomic Region
fci5	Small Deletion	epha4a
fci6	Indel	epha4a
fci7	Indel	epha4a
fci8	Small Deletion	epha4a
fci9	Indel	efnb3b
zdf1	Point Mutation	tp53

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
efnb3b	TALEN1-efnb3b	TALEN
epha4a	CRISPR2-epha4a	CRISPR
epha4a	CRISPR3-epha4a	CRISPR
epha4a	CRISPR4-epha4a	CRISPR
ppp1r12a	CRISPR5-ppp1r12a	CRISPR
ppp1r12a	CRISPR1-ppp1r12a	CRISPR
ppp1r12a	CRISPR2-ppp1r12a	CRISPR
ppp1r12a	CRISPR3-ppp1r12a	CRISPR
ppp1r12a	CRISPR4-ppp1r12a	CRISPR
ppp1r12a	MO2-ppp1r12a	MRPHLNO

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Fish

Fish
efnb3b^fci9/fci9
efnb3b^fci9/fci9 + MO2-ppp1r12a
epha4a^fci5/fci5
epha4a^fci5/fci5 + MO2-ppp1r12a
epha4a^fci6/fci6
epha4a^fci6/fci6 + MO2-ppp1r12a
epha4a^fci7/fci7
epha4a^fci7/fci7 + MO2-ppp1r12a
epha4a^fci8/fci8
WT

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Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
ab1-epha4a				Rabbit
Ab1-myl-Ser19-P	polyclonal			Rabbit
Ab1-wwtr1	monoclonal	wwtr1	IgG	Rabbit

Orthology

Engineered Foreign Genes

Mapping

Cayuso et al., 2019 ZDB-PUB-190911-8 PMID:31502954

Actomyosin regulation by Eph receptor signaling couples boundary cell formation to border sharpness

Cayuso et al., 2019

ZDB-PUB-190911-8
PMID:31502954