PUBLICATION

Akt is a mediator of artery specification during zebrafish development

Authors

Zhou, W., Ghersi, J.J., Ristori, E., Semanchik, N., Prendergast, A., Zhang, R., Carneiro, P., Baldissera, G., Sessa, W.C., Nicoli, S.

ID

ZDB-PUB-240806-2

Date

2024

Source

Development (Cambridge, England) 151(17): (Journal)

Registered Authors

Ghersi, Joey, Nicoli, Stefania, Prendergast, Andrew, Ristori, Emma

Keywords

Akt and Notch signaling, Artery specification, Endothelial cells, Single cell RNA sequencing., Vascular development, Zebrafish embryo

Datasets

GEO:GSE252648

MeSH Terms

Signal Transduction
Arteries*/embryology
Arteries*/metabolism
Animals
Mutation/genetics
Receptors, Notch*/genetics
Receptors, Notch*/metabolism
Gene Expression Regulation, Developmental
Endothelial Cells/metabolism
Zebrafish Proteins*/genetics
Zebrafish Proteins*/metabolism
Vascular Endothelial Growth Factor A/genetics
Vascular Endothelial Growth Factor A/metabolism
Embryo, Nonmammalian/metabolism
Zebrafish*/embryology
Zebrafish*/genetics
Zebrafish*/metabolism
Proto-Oncogene Proteins c-akt*/genetics
Proto-Oncogene Proteins c-akt*/metabolism

(all 19)

PubMed

39101673 Full text @ Development

Abstract

The dorsal aorta (DA) is the first major blood vessel to develop in the embryonic cardiovascular system. Its formation is governed by a coordinated process involving the migration, specification, and arrangement of angioblasts into arterial and venous lineages, a process conserved across species. While vascular endothelial growth factor a (VEGF-A) drives DA specification and formation, the kinases involved in this process remain ambiguous. Thus, we investigated the role of protein kinase B, Akt, in zebrafish by generating a quadruple mutant (aktΔ/Δ), where expression and activity of all akt genes-akt 1, 2, 3a, and 3b are strongly decreased. Live imaging of developing aktΔ/Δ DA uncovers early arteriovenous malformations. Single-cell RNA sequencing analysis of aktΔ/Δ endothelial cells corroborates the impairment of arterial, yet not venous, cell specification. Notably, endothelial specific expression of ligand-independent activation of Notch or constitutively active Akt1 were sufficient to reestablish normal arterial specification in aktΔ/Δ. The Akt-loss-of-function mutant unveils that Akt kinase can act upstream of Notch in arterial endothelial cells, and is involved in proper embryonic artery specification. This sheds light on cardiovascular development, revealing a mechanism behind congenital malformations.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
akt1	akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Day 4	whole organism	RTPCR	Fig. 1
akt2	akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Day 4	whole organism	RTPCR	Fig. 1
akt3a	akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Day 4	whole organism	RTPCR	Fig. 1
akt3b	akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Day 4	whole organism	RTPCR	Fig. 1
dlc	akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Prim-5	artery	RNA-Seq	Fig. 4
dlc	akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Prim-5	artery	ISH	Fig. 5
dlc	akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Prim-5	dorsal aorta	ISH	Fig. 5
dlc	akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Prim-5	dorsal aorta	ISH	Fig. 3
dlc	akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Prim-5	intersegmental vessel	ISH	Fig. 5
dlc	akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Prim-5	posterior cardinal vein	ISH	Fig. 3

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Prim-5	artery dlc expression decreased amount, abnormal	Fig. 4
akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Prim-5	artery dlc expression decreased amount, abnormal	Fig. 5
akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Prim-5	artery efnb2a expression decreased amount, abnormal	Fig. 4
akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Prim-5	artery notch3 expression decreased amount, abnormal	Fig. 3
akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Prim-5	artery vegfc expression decreased amount, abnormal	Fig. 4
akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Prim-5	artery endothelial cell fate specification decreased process quality, abnormal	Fig. 4
akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Prim-5	artery Notch signaling pathway decreased process quality, abnormal	Fig. 5
akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Prim-5	dorsal aorta dlc expression decreased amount, abnormal	Fig. 3
akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Prim-5	dorsal aorta dlc expression decreased amount, abnormal	Fig. 5
akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349	standard conditions	Prim-5	dorsal aorta efnb2a expression decreased amount, abnormal	Fig. 3

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
hu4881Tg	Tg(flt4:YFP)	Transgenic Insertion
s916Tg	Tg(kdrl:Hsa.HRAS-mCherry)	Transgenic Insertion
ya348		Small Deletion	akt1
ya349		Insertion	akt2
ya350		Small Deletion	akt3a
ya351		Small Deletion	akt3b

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
akt1	CRISPR2-akt1	CRISPR
akt2	CRISPR2-akt2	CRISPR
akt3a	CRISPR6-akt3a	CRISPR
akt3b	CRISPR7-akt3b	CRISPR

Fish

Fish
akt1^ya348/ya348
akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349
akt1^ya348/ya348; akt3a^ya350/ya350; akt3b^ya351/ya351; akt2^ya349/ya349; hu4881Tg/hu4881Tg; s916Tg/s916Tg
akt2^ya349/ya349
akt3a^ya350/ya350
akt3b^ya351/ya351

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Antibodies

Name	Type	Isotypes	Host Organism
Ab7-akt	monoclonal		Rabbit
Ab11-mapk	monoclonal	IgG	Rabbit
Ab15-akt	monoclonal	IgG1	Mouse

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
mCherry	EFG	mCherry
YFP	EFG	YFP

Mapping

Zhou et al., 2024 ZDB-PUB-240806-2 PMID:39101673

Akt is a mediator of artery specification during zebrafish development

Zhou et al., 2024

ZDB-PUB-240806-2
PMID:39101673