PUBLICATION

Dual role of Jam3b in early hematopoietic and vascular development

Authors

Kobayashi, I., Kobayashi-Sun, J., Hirakawa, Y., Ouchi, M., Yasuda, K., Kamei, H., Fukuhara, S., Yamaguchi, M.

ID

ZDB-PUB-191220-3

Date

2019

Source

Development (Cambridge, England) 147(1): (Journal)

Registered Authors

Fukuhara, Shigetomo, Kobayashi, Isao

Keywords

Hematopoietic stem cells, Junctional adhesion molecule, Mesoderm, Vascular endothelial cells, Zebrafish

MeSH Terms

Cardiovascular System/cytology
Cardiovascular System/embryology*
Hematopoietic Stem Cells
Receptors, Cell Surface/genetics
Receptors, Cell Surface/physiology*
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism
Zebrafish Proteins/physiology*
Endothelial Cells/physiology
Basic Helix-Loop-Helix Transcription Factors/metabolism
Animals
Hematopoiesis*/physiology
Mesoderm/embryology
Zebrafish
MAP Kinase Signaling System

(all 15)

PubMed

31852685 Full text @ Development

Abstract

In order to efficiently derive hematopoietic stem cells (HSCs) from pluripotent precursors, it is crucial to understand how mesodermal cells acquire hematopoietic and endothelial identities, two divergent, but closely related cell fates. Although Npas4 has been recently identified as a conserved master regulator of hemato-vascular development, the molecular mechanisms underlying cell fate divergence between hematopoietic and vascular endothelial cells are still unclear. Here, we show in zebrafish that mesodermal cell differentiation into hematopoietic and vascular endothelial cells is regulated by Junctional adhesion molecule 3b (Jam3b) via two independent signaling pathways. Mutation of jam3b led to a reduction in npas4l expression in the posterior lateral plate mesoderm and defects in both hematopoietic and vascular development. Mechanistically, we uncover that Jam3b promotes endothelial specification by regulating npas4l expression through repression of the Rap1a-Erk signaling cascade. Jam3b subsequently promotes hematopoietic development, including HSCs, by regulating lrrc15 expression in endothelial precursors through the activation of an integrin-dependent signaling cascade. Our data provide insight into the divergent mechanisms for instructing hematopoietic or vascular fates from mesodermal cells.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Qualifier	Anatomy	Assay	Figure
alas2	jam3b^sa37/sa37	standard conditions	10-13 somites	Not Detected	posterior lateral plate mesoderm	ISH	Fig. S2
alas2	WT	standard conditions	10-13 somites		posterior lateral plate mesoderm	ISH	Fig. S2
cdx4	jam3b^sa37/sa37	standard conditions	10-13 somites		posterior lateral plate mesoderm	ISH	Fig. S2
cdx4	WT	standard conditions	10-13 somites		posterior lateral plate mesoderm	ISH	Fig. S2
dlc	jam3b^sa37/sa37	standard conditions	10-13 somites		somite	ISH	Fig. S2
dlc	WT	control	10-13 somites		somite	ISH	Fig. S5
dlc	WT	chemical treatment: SL-327	10-13 somites		somite	ISH	Fig. S5
dlc	WT	standard conditions	10-13 somites		somite	ISH	Fig. S2
dlc	WT	chemical treatment: 2-(2-amino-3-methoxyphenyl)chromen-4-one	10-13 somites		somite	ISH	Fig. S5
drl	jam3b^sa37/sa37	chemical treatment: SL-327	10-13 somites		posterior lateral plate mesoderm	ISH	Fig. S5

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	chemical treatment: 8-(4-chlorophenylthio)-cAMP	10-13 somites	posterior lateral plate mesoderm drl expression decreased amount, abnormal	Fig. S3
WT	chemical treatment: 8-(4-chlorophenylthio)-cAMP	10-13 somites	posterior lateral plate mesoderm npas4l expression decreased amount, abnormal	Fig. S5
WT	chemical treatment: 8-(4-chlorophenylthio)-cAMP	Prim-5	dorsal aorta fli1 expression decreased distribution, abnormal	Fig. S5
WT	chemical treatment: 8-(4-chlorophenylthio)-cAMP	Prim-5	dorsal aorta runx1 expression decreased distribution, abnormal	Fig. S5
WT	chemical treatment: wortmannin	20-25 somites	vascular cord lrrc15 expression decreased amount, abnormal	Fig. 6
WT + MO1-jam3b	standard conditions	Prim-15	dorsal aorta fli1 expression decreased distribution, abnormal	Fig. S1
WT + MO1-jam3b	standard conditions	Prim-15	dorsal aorta gata1a expression decreased distribution, abnormal	Fig. S1
WT + MO1-jam3b	standard conditions	Prim-15	dorsal aorta runx1 expression decreased distribution, abnormal	Fig. S1
WT + MO1-lrrc15	standard conditions	10-13 somites	posterior lateral plate mesoderm npas4l expression decreased amount, abnormal	Fig. 3
WT + MO1-lrrc15	standard conditions	26+ somites	dorsal aorta fli1 expression decreased distribution, abnormal	Fig. 3

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
ko112Tg	Tg(UAS:mCherry-DNitgb1a)	Transgenic Insertion
kz1		Indel	lrrc15
kz2		Unknown	drl
la2Tg	Tg(-6.0itga2b:EGFP)	Transgenic Insertion
s896Tg	Tg(kdrl:Hsa.HRAS-mCherry)	Transgenic Insertion
sa37		Point Mutation	jam3b
ubs4Tg	Tg(fli1:GAL4FF)	Transgenic Insertion
y1Tg	Tg(fli1:EGFP)	Transgenic Insertion

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
drl	CRISPR1-drl,drll.1,drll.2	CRISPR
drl	MO3-drl,drll.1,drll.2	MRPHLNO
drll.1	CRISPR1-drl,drll.1,drll.2	CRISPR
drll.1	MO3-drl,drll.1,drll.2	MRPHLNO
drll.2	CRISPR1-drl,drll.1,drll.2	CRISPR
drll.2	MO3-drl,drll.1,drll.2	MRPHLNO
jam2a	MO2-jam2a	MRPHLNO
jam2b	MO1-jam2b	MRPHLNO
jam3b	MO1-jam3b	MRPHLNO
lrrc15	CRISPR1-lrrc15	CRISPR

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Fish

Fish
drl^kz2/kz2
jam3b^sa37/sa37
jam3b^sa37/sa37; la2Tg; s896Tg
jam3b^sa37; y1Tg
ko112Tg
ko112Tg; ubs4Tg
la2Tg; s896Tg
lrrc15^kz1/kz1
WT
WT + MO1-jam3b

Antibodies

Name	Type	Host Organism
Ab3-tub	polyclonal	Rabbit
Ab7-mapk	polyclonal	Rabbit
Ab13-mapk	polyclonal	Rabbit

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP
GAL4FF	EFG	GAL4FF
mCherry	EFG	mCherry

Mapping

Kobayashi et al., 2019 ZDB-PUB-191220-3 PMID:31852685

Dual role of Jam3b in early hematopoietic and vascular development

Kobayashi et al., 2019

ZDB-PUB-191220-3
PMID:31852685