PUBLICATION

Fine-tune regulation of carboxypeptidase N1 controls vascular patterning during zebrafish development

Authors

Wu, T.Y., Wang, Y.S., Song, Y.C., Chen, Z.Y., Chen, Y.T., Chiu, C.C., Wu, C.Y.

ID

ZDB-PUB-170514-3

Date

2017

Source

Scientific Reports 7: 1852 (Journal)

Registered Authors

Keywords

Angiogenesis, Gene regulation

MeSH Terms

Signal Transduction
Zebrafish/embryology*
Zebrafish/genetics*
Gene Expression Regulation, Developmental*
Neovascularization, Physiologic/genetics*
Biomarkers
Animals
RNA, Messenger/genetics
Gene Knockdown Techniques
Lysine Carboxypeptidase/genetics*
Lysine Carboxypeptidase/metabolism
Protein Binding
Organ Specificity

(all 13)

PubMed

28500283 Full text @ Sci. Rep.

Abstract

Vascular development is regulated by complicated signals and molecules in vertebrates. In this study, we characterized a novel function of carboxypeptidase N1 (Cpn1) in the vasculature. We show that cpn1 mRNA is expressed in developing vessels. The knockdown of cpn1 by morpholino injection impairs the growth of intersegmental vessels (ISV) and caudal vein plexus (CVP), suggesting the role of cpn1 in vascular development. We showed that vascular defects are not caused by cell death but are due to the impairment of migration and proliferation. Consistent with vascular growth defects, loss of cpn1 affects the expression of the vascular markers flt4, mrc1, flk, stabilin, and ephrinb2. Furthermore, the overexpression of cpn1 impaired the growth of ISV and CVP, but the remodeling expression of vascular markers was different from the knockdown of cpn1, indicating the differential regulation mechanisms in cpn1-overexpressing embryos. We examine the interaction between cpn1 and multiple signals and observed that cpn1 is regulated by Notch/VEGF signals for ISV growth and likely regulates BMP signals for CVP patterning. In conclusion, we demonstrate that cpn1 has a critical role in the vascular development of zebrafish. We also reveal a fine-tune regulation of cpn1 that controls vascular patterning mediated by multiple signals.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
cpn1	TL	control	Prim-5	blood vessel	ISH	Fig. 1
cpn1	TL	control	Prim-15	blood vessel	ISH	Fig. 1
cpn1	TL	chemical treatment by environment: DAPT	Prim-5	caudal fin	ISH	Fig. 7
cpn1	TL	chemical treatment by environment: semaxanib	Prim-5	caudal fin	ISH	Fig. 7
cpn1	TL	control	Prim-15	caudal vein plexus	ISH	Fig. 1
cpn1	TL	chemical treatment by environment: DMH1	Prim-15	caudal vein plexus	ISH	Fig. 7
cpn1	TL	control	Prim-5	caudal vein plexus	ISH	Fig. 1
cpn1	TL	control	Prim-5	caudal vein plexus	ISH	Fig. 7
cpn1	TL	control	Prim-15	caudal vein plexus	ISH	Fig. 7
cpn1	TL	chemical treatment by environment: dorsomorphin	Prim-15	caudal vein plexus	ISH	Fig. 7

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
TL	chemical treatment by environment: DAPT	Prim-5	caudal fin cpn1 expression mislocalised, abnormal	Fig. 7
TL	chemical treatment by environment: DAPT	Prim-5	caudal vein plexus cpn1 expression decreased amount, abnormal	Fig. 7
TL	chemical treatment by environment: DAPT	Prim-5	trunk vasculature cpn1 expression decreased amount, abnormal	Fig. 7
TL	chemical treatment by environment: semaxanib	Prim-5	caudal fin cpn1 expression mislocalised, abnormal	Fig. 7
TL	chemical treatment by environment: semaxanib	Prim-5	caudal vein plexus cpn1 expression decreased amount, abnormal	Fig. 7
TL	chemical treatment by environment: semaxanib	Prim-5	trunk vasculature cpn1 expression decreased amount, abnormal	Fig. 7
TL + MO3-cpn1	control	Prim-5	caudal vein plexus flt4 expression decreased amount, abnormal	Fig. 4
TL + MO3-cpn1	control	Prim-5	caudal vein plexus kdrl expression decreased amount, abnormal	Fig. 4
TL + MO3-cpn1	control	Prim-5	caudal vein plexus mrc1a expression decreased amount, abnormal	Fig. 4
TL + MO3-cpn1	control	Prim-5	caudal vein plexus stab2 expression decreased amount, abnormal	Fig. 4

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Mutations / Transgenics

Allele	Construct	Type
ci5Tg	Tg(-6.5kdrl:mCherry)	Transgenic Insertion
la116Tg	Tg(kdrl:GFP)	Transgenic Insertion
sd2Tg	Tg(gata1a:DsRed)	Transgenic Insertion
y1Tg	Tg(fli1:EGFP)	Transgenic Insertion
y7Tg	Tg(fli1:nEGFP)	Transgenic Insertion

1 - 5 of 5

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
cpn1	MO3-cpn1	MRPHLNO
cpn1	MO4-cpn1	MRPHLNO
tp53	MO4-tp53	MRPHLNO

Fish

Fish
ci5Tg; y7Tg
ci5Tg; y7Tg + MO3-cpn1
la116Tg + MO3-cpn1
sd2Tg; y1Tg
sd2Tg; y1Tg + MO3-cpn1
TL
TL + MO3-cpn1
y1Tg
y1Tg + MO3-cpn1

Antibodies

Orthology

Gene	Orthology
cpn1

Engineered Foreign Genes

Marker	Marker Type	Name
DsRed	EFG	DsRed
EGFP	EFG	EGFP
GFP	EFG	GFP
mCherry	EFG	mCherry

Mapping

Wu et al., 2017 ZDB-PUB-170514-3 PMID:28500283

Fine-tune regulation of carboxypeptidase N1 controls vascular patterning during zebrafish development

Wu et al., 2017

ZDB-PUB-170514-3
PMID:28500283