PUBLICATION

RPTPalpha and PTPepsilon Signaling via Fyn/Yes and RhoA is Essential for Zebrafish Convergence and Extension Cell Movements during Gastrulation

Authors

van Eekelen, M., Runtuwene, V., Overvoorde, J., and den Hertog, J.

ID

ZDB-PUB-100302-20

Date

2010

Source

Developmental Biology 340(2): 626-639 (Journal)

Registered Authors

den Hertog, Jeroen, Overvoorde, John

Keywords

Zebrafish, Protein-tyrosine phosphatase, RPTPα, PTPε, Src family kinases, Convergence and extension cell movements, Gastrulation

MeSH Terms

Cell Movement/physiology
Gastrulation/physiology*
Zebrafish/genetics
Zebrafish/metabolism
Zebrafish/physiology*
Zebrafish Proteins/genetics
Zebrafish Proteins/physiology*
Embryo, Nonmammalian/metabolism
Embryo, Nonmammalian/physiology
Receptor-Like Protein Tyrosine Phosphatases, Class 4/physiology*
Monomeric GTP-Binding Proteins/genetics
Monomeric GTP-Binding Proteins/physiology*
rhoA GTP-Binding Protein/genetics
rhoA GTP-Binding Protein/physiology*
Proto-Oncogene Proteins c-fyn/genetics
Proto-Oncogene Proteins c-fyn/physiology*
Animals
Signal Transduction/physiology

(all 18)

PubMed

20188722 Full text @ Dev. Biol.

Abstract

Convergence and extension (C&E) cell movements are essential to shape the body axis during vertebrate gastrulation. We have used the zebrafish to assess the role of the receptor protein-tyrosine phosphatases, RPTPalpha and PTPepsilon, in gastrulation cell movements. Both RPTPalpha and PTPepsilon knockdown and ptpra(-/-) embryos show defects in C&E movements. A method was developed to track gastrulation cell movements using confocal microscopy in a quantitative manner and ptpra(-/-) embryos displayed reduced convergence as well as extension speeds. RPTPalpha and PTPepsilon knockdowns cooperated with knockdown of a well known factor in C&E cell movement, non-canonical Wnt11. RPTPalpha and PTPepsilon dephosphorylate and activate Src family kinases in various cell types in vitro and in vivo. We found that Src family kinase phosphorylation was enhanced in ptpra(-/-) embryos, consistent with reduced Src family kinase activity. Importantly, both ptpra(-/-) as well as RPTPalpha and PTPepsilon knockdown induced C&E defects were rescued by active Fyn and Yes. Moreover, active RhoA rescued the RPTPalpha and PTPepsilon knockdown and ptpra(-/-) induced gastrulation cell movement defects as well. Our results demonstrate that RPTPalpha and PTPepsilon are essential for C&E movements in a signaling pathway parallel to non-canonical Wnts and upstream of Fyn, Yes and RhoA.

Genes / Markers

Figures

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Expression

Gene	Antibody	Fish	Conditions	Stage	Qualifier	Anatomy	Assay	Figure
ptpra	ab1-ptpra	ptpra^{hu3334/hu3334}	standard conditions	Long-pec	Not Detected	whole organism	WB	Fig. 8
ptpra		WT	standard conditions	Prim-25		whole organism	ISH	Fig. S1
ptpra		WT	standard conditions	Shield		whole organism	ISH	Fig. S1
ptpra		WT	standard conditions	1-4 somites		whole organism	ISH	Fig. S1
ptpra		WT	standard conditions	8-cell		whole organism	ISH	Fig. S1
ptpra		WT	standard conditions	Protruding-mouth		whole organism	ISH	Fig. S1
ptpra		WT	standard conditions	Prim-5		whole organism	ISH	Fig. S1
ptpra	ab1-ptpra	WT	standard conditions	Long-pec		whole organism	WB	Fig. 8
ptprea		WT	standard conditions	Shield		whole organism	ISH	Fig. S1
ptprea		WT	standard conditions	1-4 somites		whole organism	ISH	Fig. S1

1 - 10 of 20

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT + MO1-ptpra	standard conditions	Long-pec	convergent extension involved in gastrulation disrupted, abnormal	Fig. 2
WT + MO1-ptpra	standard conditions	Long-pec	extension decreased length, abnormal	Fig. 2
WT + MO1-ptpra	standard conditions	Long-pec	pericardium edematous, abnormal	Fig. 2
WT + MO1-ptpra	standard conditions	Long-pec	whole organism decreased length, abnormal	Fig. 2
WT + MO1-ptpra	standard conditions	Long-pec	yolk edematous, abnormal	Fig. 2
WT + MO1-ptpra + MO1-ptprea + MO1-ptpreb	standard conditions	Long-pec	convergent extension involved in gastrulation disrupted, abnormal	Fig. 2
WT + MO1-ptpra + MO1-ptprea + MO1-ptpreb	standard conditions	Long-pec	eye fused with eye, abnormal	Fig. 2
WT + MO1-ptpra + MO1-ptprea + MO1-ptpreb	standard conditions	Long-pec	pericardium edematous, abnormal	Fig. 2
WT + MO1-ptpra + MO1-ptprea + MO1-ptpreb	standard conditions	Long-pec	whole organism decreased length, abnormal	Fig. 2
WT + MO1-ptpra + MO1-wnt11f2	standard conditions	Protruding-mouth	eye fused with eye, abnormal	Fig. 7

1 - 10 of 38

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
hu3334		Point Mutation	ptpra

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
ptpra	MO1-ptpra	MRPHLNO
ptprea	MO1-ptprea	MRPHLNO
ptpreb	MO1-ptpreb	MRPHLNO
wnt11f2	MO1-wnt11f2	MRPHLNO

Fish

Fish
ptpra^{hu3334/hu3334}
WT
WT + MO1-ptpra
WT + MO1-ptpra + MO1-ptprea + MO1-ptpreb
WT + MO1-ptpra + MO1-wnt11f2
WT + MO1-ptprea + MO1-ptpreb
WT + MO1-ptprea + MO1-ptpreb + MO1-wnt11f2
WT + MO1-wnt11f2

1 - 8 of 8

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Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab1-ptpra	polyclonal	ptpra		Rabbit

Orthology

Engineered Foreign Genes

Mapping

van Eekelen et al., 2010 ZDB-PUB-100302-20 PMID:20188722

RPTPalpha and PTPepsilon Signaling via Fyn/Yes and RhoA is Essential for Zebrafish Convergence and Extension Cell Movements during Gastrulation

van Eekelen et al., 2010

ZDB-PUB-100302-20
PMID:20188722