PUBLICATION

The Cell Adhesion-associated Protein Git2 Regulates Morphogenetic Movements during Zebrafish Embryonic Development

Authors

Yu, J.A., Foley, F.C., Amack, J.D., and Turner, C.E.

ID

ZDB-PUB-101108-1

Date

2011

Source

Developmental Biology 349(2): 225-237 (Journal)

Registered Authors

Amack, Jeffrey, Foley, Fiona

Keywords

Zebrafish, Epiboly, Gastrulation, Git2, Non-muscle Myosin II, Blebbistatin, Paxillin, FAK

MeSH Terms

Heterocyclic Compounds, 4 or More Rings
Phylogeny
GTPase-Activating Proteins/genetics
GTPase-Activating Proteins/metabolism*
Cell Movement/physiology*
Molecular Sequence Data
Time-Lapse Imaging
In Situ Hybridization
Morphogenesis/physiology*
Cell Adhesion Molecules/metabolism*
Embryonic Development/physiology*
Zebrafish/embryology*
Sequence Analysis, DNA
Immunohistochemistry
Gene Knockdown Techniques
Base Sequence
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism
Phosphorylation
Immunoblotting
Signal Transduction/physiology*
Myosin Light Chains/metabolism
Animals

(all 23)

PubMed

21034731 Full text @ Dev. Biol.

Abstract

Signaling through cell adhesion complexes plays a critical role in coordinating cytoskeletal remodeling necessary for efficient cell migration. During embryonic development, normal morphogenesis depends on a series of concerted cell movements; but the roles of cell adhesion signaling during these movements are poorly understood. The transparent zebrafish embryo provides an excellent system to study cell migration during development. Here, we have identified zebrafish git2a and git2b, two new members of the GIT family of genes that encode ArfGAP proteins associated with cell adhesions. Loss-of-function studies revealed an essential role for Git2a in zebrafish cell movements during gastrulation. Time-lapse microscopy analysis demonstrated that antisense depletion of Git2a greatly reduced or arrested cell migration towards the vegetal pole of the embryo. These defects were rescued by expression of chicken GIT2, indicating a specific and conserved role for Git2 in controlling embryonic cell movements. Git2a knockdown embryos showed defects in cell morphology that were associated with reduced cell contractility. We show that Git2a is required for phosphorylation of myosin light chain (MLC), which regulates myosin II-mediated cell contractility. Consistent with this, embryos treated with Blebbistatin-a small molecule inhibitor for myosin II activity-exhibited cell movement defects similar to git2a knockdown embryos. These observations provide in vivo evidence of a physiologic role for Git2a in regulating cell morphogenesis and directed cell migration via myosin II activation during zebrafish embryonic development.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
git2a	AB	standard conditions	Prim-5	whole organism	ISH	Fig. 1
git2a	AB	standard conditions	4-cell	whole organism	ISH	Fig. 1
git2a	AB	standard conditions	Bud	whole organism	ISH	Fig. 1
git2a	AB	standard conditions	75%-epiboly	whole organism	ISH	Fig. 1
git2a	AB	standard conditions	14-19 somites	whole organism	ISH	Fig. 1
git2b	AB	standard conditions	Prim-5	whole organism	ISH	Fig. S2
git2b	AB	standard conditions	4-cell	whole organism	ISH	Fig. S2
git2b	AB	standard conditions	Bud	whole organism	ISH	Fig. S2
git2b	AB	standard conditions	30%-epiboly to 50%-epiboly	whole organism	ISH	Fig. S2
git2b	AB	standard conditions	14-19 somites	whole organism	ISH	Fig. S2

1 - 10 of 10

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
AB + MO1-git2a	standard conditions	50%-epiboly	cell migration involved in gastrulation process quality, abnormal	Fig. 3
AB + MO1-git2a	standard conditions	50%-epiboly	cell migration involved in gastrulation rate, normal	Fig. 3
AB + MO1-git2a	standard conditions	Shield	epiboly involved in gastrulation with mouth forming second disrupted, abnormal	Fig. 2
AB + MO1-git2a	standard conditions	Shield to 90%-epiboly	cell migration involved in gastrulation decreased rate, abnormal	Fig. 3
AB + MO1-git2a	standard conditions	Shield to 90%-epiboly	cell migration involved in gastrulation disrupted, abnormal	Fig. 3
AB + MO1-git2a	standard conditions	Shield to 90%-epiboly	epiboly involved in gastrulation with mouth forming second disrupted, abnormal	Fig. 3
AB + MO1-git2a	standard conditions	75%-epiboly	EVL adherens junction composition, normal	Fig. 6
AB + MO1-git2a	standard conditions	75%-epiboly	EVL cell morphology, abnormal	Fig. 4
AB + MO1-git2a	standard conditions	75%-epiboly	EVL cell morphology, abnormal	Fig. 6
AB + MO1-git2a	standard conditions	90%-epiboly	embryo development delayed, abnormal	Fig. 2

1 - 10 of 21

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Mutations / Transgenics

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
git2a	MO1-git2a	MRPHLNO
git2a	MO2-git2a	MRPHLNO

Fish

Fish
AB
AB + MO1-git2a
AB + MO2-git2a

Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab1-git	monoclonal		IgG1	Mouse
Ab1-myl-Ser19-P	polyclonal			Rabbit

Orthology

Gene	Orthology
git2a
git2b

Engineered Foreign Genes

Mapping

Yu et al., 2011 ZDB-PUB-101108-1 PMID:21034731

The Cell Adhesion-associated Protein Git2 Regulates Morphogenetic Movements during Zebrafish Embryonic Development

Yu et al., 2011

ZDB-PUB-101108-1
PMID:21034731