PUBLICATION

The recycling endosome protein Rab25 coordinates collective cell movements in the zebrafish surface epithelium

Authors

Willoughby, P.M., Allen, M., Yu, J., Korytnikov, R., Chen, T., Liu, Y., So, I., Macpherson, N., Mitchell, J.A., Fernandez-Gonzalez, R., Bruce, A.E.E.

ID

ZDB-PUB-210324-8

Date

2021

Source

eLIFE 10: (Journal)

Registered Authors

Bruce, Ashley

Keywords

cell biology, developmental biology, zebrafish

MeSH Terms

Cytokinesis
Zebrafish/genetics
Zebrafish/physiology*
Gastrula/physiology
Animals
Embryo, Nonmammalian/physiology
rab GTP-Binding Proteins/genetics*
rab GTP-Binding Proteins/metabolism
Cell Movement/genetics*
Zebrafish Proteins
Epithelium/physiology

(all 11)

PubMed

33755014 Full text @ Elife

Abstract

In emerging epithelial tissues, cells undergo dramatic rearrangements to promote tissue shape changes. Dividing cells remain interconnected via transient cytokinetic bridges. Bridges are cleaved during abscission and currently, the consequences of disrupting abscission in developing epithelia are not well understood. We show that the Rab GTPase, Rab25, localizes near cytokinetic midbodies and likely coordinates abscission through endomembrane trafficking in the epithelium of the zebrafish gastrula during epiboly. In maternal-zygotic Rab25a and Rab25b mutant embryos, morphogenic activity tears open persistent apical cytokinetic bridges that failed to undergo timely abscission. Cytokinesis defects result in anisotropic cell morphologies that are associated with a reduction of contractile actomyosin networks. This slows cell rearrangements and alters the viscoelastic responses of the tissue, all of which likely contribute to delayed epiboly. We present a model in which Rab25 trafficking coordinates cytokinetic bridge abscission and cortical actin density, impacting local cell shape changes and tissue-scale forces.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
gsc	rab25a^uot10/uot10	standard conditions	75%-epiboly	mesoderm	ISH	Figure 2—figure supplement 1.
gsc	rab25a^uot9/uot9	standard conditions	75%-epiboly	mesoderm	ISH	Figure 2—figure supplement 1.
gsc	rab25b^uot11/uot11	standard conditions	75%-epiboly	mesoderm	ISH	Figure 2—figure supplement 1.
gsc	WT	standard conditions	75%-epiboly	mesoderm	ISH	Figure 2—figure supplement 1.
krt4	rab25a^uot10/uot10	standard conditions	Shield	EVL	ISH	Figure 2—figure supplement 1.
krt4	rab25a^uot9/uot9	standard conditions	Shield	EVL	ISH	Figure 2—figure supplement 1.
krt4	rab25b^uot11/uot11	standard conditions	Shield	EVL	ISH	Figure 2—figure supplement 1.
krt4	WT	standard conditions	Shield	EVL	ISH	Figure 2—figure supplement 1.
rab11a	rab25a^uot10/uot10	standard conditions	Shield	whole organism	RTPCR	Figure 2—figure supplement 1.
rab11a	rab25a^uot9/uot9	standard conditions	Shield	whole organism	RTPCR	Figure 2—figure supplement 1.

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
rab25a^uot10/uot10	control	75%-epiboly	EVL endocytic recycling decreased process quality, abnormal	Figure 7
rab25a^uot10/uot10	standard conditions	1k-cell	blastomere morphology, abnormal	Figure 2—figure supplement 1.
rab25a^uot10/uot10	standard conditions	Dome	epiboly onset quality, normal	Figure 2
rab25a^uot10/uot10	standard conditions	Sphere	EVL cell morphology, normal	Figure 2—figure supplement 2.
rab25a^uot10/uot10	standard conditions	Shield	EVL cell morphology, abnormal	Figure 2—figure supplement 2.
rab25a^uot10/uot10	standard conditions	Shield	shield morphology, normal	Figure 2
rab25a^uot10/uot10	standard conditions	Shield	whole organism rab25a expression decreased amount, abnormal	Figure 2—figure supplement 1.
rab25a^uot10/uot10	standard conditions	Shield	whole organism rab25b expression increased amount, abnormal	Figure 2—figure supplement 1.
rab25a^uot10/uot10	standard conditions	Shield to 90%-epiboly	epiboly delayed, abnormal	Figure 2
rab25a^uot10/uot10	standard conditions	75%-epiboly	DEL epiboly delayed, abnormal	Figure 2—figure supplement 1.

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
e2212Tg	Tg(actb2:myl12.1-EGFP)	Transgenic Insertion
io007Tg	Tg(Xla.Eef1a1:dclk2DeltaK-GFP)	Transgenic Insertion
uot3Tg	Tg(Xla.Eef1a1:EGFP-tuba5)	Transgenic Insertion
uot10		Indel	rab25a
uot11		Small Deletion	rab25b
uot16Tg	Tg(actb1:rab25b,myl7:RFP)	Transgenic Insertion
uot9		Small Deletion	rab25a

1 - 7 of 7

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
rab25a	CRISPR1-rab25a	CRISPR
rab25b	CRISPR1-rab25b	CRISPR

Fish

Fish
AB
rab25a^uot9/uot9
rab25a^uot10/uot10
rab25b^uot11/uot11
rab25b^uot11/uot11; uot16Tg
WT

Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab1-myl-Ser19-P	polyclonal			Rabbit
Ab3-cdh1	polyclonal		IgG	Rabbit

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP
GFP	EFG	GFP
RFP	EFG	RFP

Mapping

Willoughby et al., 2021 ZDB-PUB-210324-8 PMID:33755014

The recycling endosome protein Rab25 coordinates collective cell movements in the zebrafish surface epithelium

Willoughby et al., 2021

ZDB-PUB-210324-8
PMID:33755014