PUBLICATION

The Ribosome Biogenesis Protein Nol9 Is Essential for Definitive Hematopoiesis and Pancreas Morphogenesis in Zebrafish

Authors

Bielczyk-Maczyńska, E., Lam Hung, L., Ferreira, L., Fleischmann, T., Weis, F., Fernández-Pevida, A., Harvey, S.A., Wali, N., Warren, A.J., Barroso, I., Stemple, D.L., Cvejic, A.

ID

ZDB-PUB-151202-3

Date

2015

Source

PLoS Genetics 11: e1005677 (Journal)

Registered Authors

Stemple, Derek L.

Keywords

Larvae, Pancreas, Embryos, Zebrafish, Ribosomal RNA, Ribosomes, Biosynthesis, In situ hybridization

MeSH Terms

Hematopoiesis/genetics
Zebrafish
Hematopoietic Stem Cells/pathology
Morphogenesis/genetics*
Pancreas/metabolism
Pancreas/pathology
RNA, Ribosomal, 28S/genetics
Animals
Humans
Tumor Suppressor Protein p53/genetics*
Disease Models, Animal
Ribosomes/genetics*
Ribosomes/pathology
Polynucleotide 5'-Hydroxyl-Kinase/biosynthesis*
Polynucleotide 5'-Hydroxyl-Kinase/genetics

(all 15)

PubMed

26624285 Full text @ PLoS Genet.

Abstract

Ribosome biogenesis is a ubiquitous and essential process in cells. Defects in ribosome biogenesis and function result in a group of human disorders, collectively known as ribosomopathies. In this study, we describe a zebrafish mutant with a loss-of-function mutation in nol9, a gene that encodes a non-ribosomal protein involved in rRNA processing. nol9sa1022/sa1022 mutants have a defect in 28S rRNA processing. The nol9sa1022/sa1022 larvae display hypoplastic pancreas, liver and intestine and have decreased numbers of hematopoietic stem and progenitor cells (HSPCs), as well as definitive erythrocytes and lymphocytes. In addition, ultrastructural analysis revealed signs of pathological processes occurring in endothelial cells of the caudal vein, emphasizing the complexity of the phenotype observed in nol9sa1022/sa1022 larvae. We further show that both the pancreatic and hematopoietic deficiencies in nol9sa1022/sa1022 embryos were due to impaired cell proliferation of respective progenitor cells. Interestingly, genetic loss of Tp53 rescued the HSPCs but not the pancreatic defects. In contrast, activation of mRNA translation via the mTOR pathway by L-Leucine treatment did not revert the erythroid or pancreatic defects. Together, we present the nol9sa1022/sa1022 mutant, a novel zebrafish ribosomopathy model, which recapitulates key human disease characteristics. The use of this genetically tractable model will enhance our understanding of the tissue-specific mechanisms following impaired ribosome biogenesis in the context of an intact vertebrate.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Qualifier	Anatomy	Assay	Figure
EGFP	jh1Tg; jh2Tg	standard conditions	Day 4		exocrine pancreas	IFL	Fig. 3
EGFP	jh1Tg; jh2Tg	control	Day 5		exocrine pancreas	IFL	Fig. S5
EGFP	jh1Tg; jh2Tg + MO1-nol9	control	Day 5		exocrine pancreas	IFL	Fig. S5
EGFP	nol9^{sa1022/sa1022}; jh1Tg; jh2Tg	standard conditions	Day 4		exocrine pancreas	IFL	Fig. 3
foxn1	nol9^sa1022/+	standard conditions	Day 4		thymic epithelium	ISH	Fig. 4
foxn1	WT	standard conditions	Day 4		thymic epithelium	ISH	Fig. 4
hbae1.1	nol9^{sa1022/sa1022}	standard conditions	Day 5	Not Detected	hematopoietic system nucleate erythrocyte	ISH	Fig. 1
hbae1.1	nol9^{sa1022/sa1022}	standard conditions	Day 4		post-vent region erythroid progenitor cell	ISH	Fig. 7
hbae1.1	nol9^{sa1022/sa1022}; tp53^zdf1/+	standard conditions	Day 4		post-vent region erythroid progenitor cell	ISH	Fig. 7
hbae1.1	nol9^{sa1022/sa1022}; tp53^zdf1/zdf1	standard conditions	Day 4		post-vent region erythroid progenitor cell	ISH	Fig. 7

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
jh1Tg; jh2Tg + MO1-nol9	control	Day 5	endocrine pancreas size, normal	Fig. S5
jh1Tg; jh2Tg + MO1-nol9	control	Day 5	exocrine pancreas decreased size, abnormal	Fig. S5
jh1Tg; jh2Tg + MO1-nol9	control	Day 5	exocrine pancreas EGFP expression decreased amount, abnormal	Fig. S5
nol9^{sa1022/sa1022}	standard conditions	Protruding-mouth	caudal hematopoietic tissue tp53 expression increased amount, abnormal	Fig. 8
nol9^{sa1022/sa1022}	standard conditions	Protruding-mouth	intestine tp53 expression increased amount, abnormal	Fig. 8
nol9^{sa1022/sa1022}	standard conditions	Protruding-mouth	liver tp53 expression increased amount, abnormal	Fig. 8
nol9^{sa1022/sa1022}	standard conditions	Protruding-mouth	post-vent region hematopoietic multipotent progenitor cell myb expression decreased amount, abnormal	Fig. 4
nol9^{sa1022/sa1022}	standard conditions	Protruding-mouth	post-vent region hematopoietic multipotent progenitor cell decreased amount, abnormal	Fig. 4
nol9^{sa1022/sa1022}	standard conditions	Day 4	hematopoietic multipotent progenitor cell decreased amount, abnormal	Fig. 7
nol9^{sa1022/sa1022}	standard conditions	Day 4	lymphocyte decreased amount, abnormal	Fig. 4

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
jh1Tg	Tg(ptf1a:EGFP)	Transgenic Insertion
jh2Tg	Tg(ins:mCherry)	Transgenic Insertion
la2Tg	Tg(-6.0itga2b:EGFP)	Transgenic Insertion
sa1022		Point Mutation	nol9
zdf1		Point Mutation	tp53
zf169Tg	TgPAC(myb:2xmyb-EGFP)	Transgenic Insertion

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
nol9	MO1-nol9	MRPHLNO

Fish

Fish
jh1Tg
jh1Tg; jh2Tg
jh1Tg; jh2Tg + MO1-nol9
nol9^{sa1022/sa1022}
nol9^{sa1022/sa1022}; jh1Tg
nol9^{sa1022/sa1022}; jh1Tg; jh2Tg
nol9^{sa1022/sa1022}; la2Tg
nol9^{sa1022/sa1022}; tp53^zdf1/+
nol9^{sa1022/sa1022}; tp53^zdf1/zdf1
nol9^{sa1022/sa1022}; zf169Tg

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Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab1-gcg	monoclonal		IgG1	Mouse
Ab1-ins	polyclonal	ins		Guinea pig
Ab2-cpa1	polyclonal			Rabbit
Ab3-sst	polyclonal			Rabbit
Ab4-krt18	monoclonal		IgG1	Mouse

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Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP
mCherry	EFG	mCherry

Mapping

Bielczyk-Maczyńska et al., 2015 ZDB-PUB-151202-3 PMID:26624285

The Ribosome Biogenesis Protein Nol9 Is Essential for Definitive Hematopoiesis and Pancreas Morphogenesis in Zebrafish

Bielczyk-Maczyńska et al., 2015

ZDB-PUB-151202-3
PMID:26624285