PUBLICATION

A Conserved Notochord Enhancer Controls Pancreas Development in Vertebrates

Authors

Amorim, J.P., Gali-Macedo, A., Marcelino, H., Bordeira-Carriço, R., Naranjo, S., Rivero-Gil, S., Teixeira, J., Galhardo, M., Marques, J., Bessa, J.

ID

ZDB-PUB-200710-12

Date

2020

Source

Cell Reports 32: 107862 (Journal)

Registered Authors

Bessa, Jose, Naranjo, Silvia

Keywords

notochord, pancreas development, signaling diffusion, transcriptional regulation, zebrafish

MeSH Terms

Animals
Conserved Sequence/genetics*
Enhancer Elements, Genetic*
Gene Expression Regulation, Developmental
Genome
Models, Biological
Notochord/embryology*
Organ Size/genetics
Pancreas/embryology*
Pancreas/metabolism
Vertebrates/embryology*
Vertebrates/genetics*
Zebrafish/embryology
Zebrafish/genetics
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism

(all 16)

PubMed

32640228 Full text @ Cell Rep.

Abstract

The notochord is an evolutionary novelty in vertebrates that functions as an important signaling center during development. Notochord ablation in chicken has demonstrated that it is crucial for pancreas development; however, the molecular mechanism has not been fully described. Here, we show that in zebrafish, the loss of function of nog2, a Bmp antagonist expressed in the notochord, impairs β cell differentiation, compatible with the antagonistic role of Bmp in β cell differentiation. In addition, we show that nog2 expression in the notochord is induced by at least one notochord enhancer and its loss of function reduces the number of pancreatic progenitors and impairs β cell differentiation. Tracing Nog2 diffusion, we show that Nog2 emanates from the notochord to the pancreas progenitor domain. Finally, we find a notochord enhancer in human and mice Nog genomic landscapes, suggesting that the acquisition of a Nog notochord enhancer occurred early in the vertebrate phylogeny and contributes to the development of complex organs like the pancreas.

Genes / Markers

Marker	Marker Type	Name
en.nog2.e2	ENHANCER	nog2 enhancer 2
en.nog2e1	ENHANCER	nog2 enhancer 1
en.nog2e3	ENHANCER	nog2E3 notochord enhancer
foxa2	GENE	forkhead box A2
gata2a	GENE	GATA binding protein 2a
ins	GENE	preproinsulin
meis1b	GENE	Meis homeobox 1 b
nog2	GENE	noggin 2

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Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
EGFP	iis10Tg/iis10Tg	standard conditions	14-19 somites	notochord	IFL	Figure 5
EGFP	iis9Tg/iis9Tg	standard conditions	14-19 somites	notochord	IFL	Figure 3
EGFP	iis9Tg/iis9Tg	standard conditions	14-19 somites	pancreas primordium	IFL	Figure 3
ins	AB + CRISPR1-en.nog2e3 + CRISPR2-en.nog2e3	standard conditions	Long-pec	pancreatic B cell	ISH	Figure 4
ins	iis9Tg/iis9Tg	standard conditions	Long-pec	pancreatic B cell	ISH	Figure 4
ins	nog2^iis1/+	standard conditions	Long-pec	pancreas	ISH	Figure 1
ins	nog2^iis1/+; nog2^iis3/iis3	standard conditions	Long-pec	pancreas	ISH	Figure 1
ins	nog2^iis3/iis3	standard conditions	Long-pec	pancreas	ISH	Figure 1

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
AB + CRISPR1-en.nog2e3 + CRISPR2-en.nog2e3	standard conditions	Long-pec	pancreatic B cell ins expression decreased amount, abnormal	Figure 4
iis10Tg/iis10Tg	standard conditions	14-19 somites	notochord EGFP expression increased distribution, abnormal	Figure 5
iis9Tg/iis9Tg	standard conditions	14-19 somites	notochord core promoter sequence-specific DNA binding process quality, normal	Figure 3
iis9Tg/iis9Tg	standard conditions	14-19 somites	notochord EGFP expression increased distribution, abnormal	Figure 3
iis9Tg/iis9Tg	standard conditions	14-19 somites	notochord pancreas induction process quality, normal	Figure 3
iis9Tg/iis9Tg	standard conditions	14-19 somites	pancreas primordium EGFP expression increased distribution, abnormal	Figure 3
iis9Tg/iis9Tg	standard conditions	Long-pec	pancreatic B cell ins expression increased amount, abnormal	Figure 4
nog2^iis1/+	standard conditions	Long-pec	pancreas ins expression decreased amount, abnormal	Figure 1
nog2^iis1/+	standard conditions	Long-pec	pancreas ab3-nkx6 labeling decreased amount, abnormal	Figure 1
nog2^iis1/+; nog2^iis3/iis3	standard conditions	Long-pec	pancreas ins expression decreased amount, abnormal	Figure 1

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
iis1		Small Deletion	nog2
iis2		Small Deletion	nog2
iis3		Indel	nog2
iis4		Indel	nog2
iis5Tg	Tg(gata2a:GFP-LOXP-GAB-5HS4,meis1b:DsRed-LOXP)	Transgenic Insertion	nog2
iis6Tg	Tg(ins:GFP)	Transgenic Insertion
iis7Tg	Tg(FRT-Xla.Actc1:DsRed-GAB-FRT,LOXP-en.nog2e1-LOXP-gata2a:EGFP-5HS4)	Transgenic Insertion
iis8Tg	Tg(FRT-Xla.Actc1:DsRed-GAB-FRT,LOXP-en.nog2.e2-LOXP-gata2a:EGFP-5HS4)	Transgenic Insertion
iis9Tg	Tg(FRT-Xla.Actc1:DsRed-GAB-FRT,LOXP-en.nog2e3-LOXP-gata2a:EGFP-5HS4)	Transgenic Insertion
iis10Tg	Tg(FRT-Xla.Actc1:DsRed-GAB-FRT,LOXP-gata2a-Hsa.NOGD10-LOXP-gata2a:EGFP-5HS4)	Transgenic Insertion

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
en.nog2e3	CRISPR1-en.nog2e3	CRISPR
en.nog2e3	CRISPR2-en.nog2e3	CRISPR
foxa2	MO1-foxa2	MRPHLNO
nog2	CRISPR1-nog2	CRISPR
nog2	MO1-nog2	MRPHLNO

Fish

Antibodies

Orthology

Engineered Foreign Genes

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

Mapping

Amorim et al., 2020 ZDB-PUB-200710-12 PMID:32640228

A Conserved Notochord Enhancer Controls Pancreas Development in Vertebrates

Amorim et al., 2020

ZDB-PUB-200710-12
PMID:32640228