PUBLICATION

Entpd5 is essential for skeletal mineralization and regulates phosphate homeostasis in zebrafish

Authors

Huitema, L.F., Apschner, A., Logister, I., Spoorendonk, K.M., Bussmann, J., Hammond, C.L., and Schulte-Merker, S.

ID

ZDB-PUB-121220-19

Date

2012

Source

Proceedings of the National Academy of Sciences of the United States of America 109(52): 21372-21377 (Journal)

Registered Authors

Apschner, Alexander, Bussmann, Jeroen, Hammond, Chrissy, Huitema, Leonie, Logister, Ive, Schulte-Merker, Stefan, Spoorendonk, Kirsten

Keywords

none

Datasets

GEO:GSE35737

MeSH Terms

Molecular Sequence Data
Bone and Bones/embryology
Bone and Bones/metabolism
Bone and Bones/pathology
Phenotype
Humans
Osteoblasts/enzymology
Zebrafish Proteins/chemistry
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism*
Base Sequence
Calcification, Physiologic*
Animals
Pyrophosphatases/chemistry
Pyrophosphatases/genetics
Pyrophosphatases/metabolism*
Alleles
Amino Acid Sequence
Organ Specificity
Zebrafish/embryology
Zebrafish/metabolism*
Phosphates/metabolism*
Phosphoric Diester Hydrolases/genetics
Embryo, Nonmammalian/metabolism
Homeostasis*
Mutation/genetics

(all 26)

PubMed

23236130 Full text @ Proc. Natl. Acad. Sci. USA

Abstract

Bone mineralization is an essential step during the embryonic development of vertebrates, and bone serves vital functions in human physiology. To systematically identify unique gene functions essential for osteogenesis, we performed a forward genetic screen in zebrafish and isolated a mutant, no bone (nob), that does not form any mineralized bone. Positional cloning of nob identified the causative gene to encode ectonucleoside triphosphate/diphosphohydrolase 5 (entpd5); analysis of its expression pattern demonstrates that entpd5 is specifically expressed in osteoblasts. An additional mutant, dragonfish (dgf), exhibits ectopic mineralization in the craniofacial and axial skeleton and encodes a loss-of-function allele of ectonucleotide pyrophosphatase phosphodiesterase 1 (enpp1). Intriguingly, generation of double-mutant nob/dgf embryos restored skeletal mineralization in nob mutants, indicating that mechanistically, Entpd5 and Enpp1 act as reciprocal regulators of phosphate/pyrophosphate homeostasis in vivo. Consistent with this, entpd5 mutant embryos can be rescued by high levels of inorganic phosphate, and phosphate-regulating factors, such as fgf23 and npt2a, are significantly affected in entpd5 mutant embryos. Our study demonstrates that Entpd5 represents a previously unappreciated essential player in phosphate homeostasis and skeletal mineralization.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
col10a1a	entpd5a^{hu3718/hu3718}	standard conditions	Protruding-mouth	osteoblast	ISH	Fig. 1
col10a1a	WT	standard conditions	Protruding-mouth	osteoblast	ISH	Fig. 1
col1a2	entpd5a^{hu3718/hu3718}	standard conditions	Protruding-mouth	osteoblast	ISH	Fig. 1
col1a2	WT	standard conditions	Protruding-mouth	osteoblast	ISH	Fig. 1
entpd5a	entpd5a^{hu3718/hu3718}	standard conditions	Days 7-13	whole organism	RTPCR	Fig. 4
entpd5a	WT	standard conditions	Protruding-mouth	cleithrum	ISH	Fig. 2
entpd5a	WT	standard conditions	Protruding-mouth	opercular flap	ISH	Fig. 2
entpd5a	WT	standard conditions	Protruding-mouth	parasphenoid	ISH	Fig. 2
entpd5a	WT	standard conditions	Days 7-13	whole organism	RTPCR	Fig. 4
fgf23	entpd5a^{hu3718/hu3718}	standard conditions	Days 7-13	whole organism	RTPCR	Fig. 4

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
enpp1^{hu4581/hu4581}	standard conditions	Days 7-13	axial fin skeleton mineralized, abnormal	Fig. 3
enpp1^{hu4581/hu4581}	standard conditions	Days 7-13	bone mineralization increased process quality, abnormal	Fig. 3
enpp1^{hu4581/hu4581}	standard conditions	Days 7-13	cleithrum nodular, abnormal	Fig. 3
enpp1^{hu4581/hu4581}	standard conditions	Days 7-13	vertebra centrum fused with centrum, abnormal	Fig. 3
enpp1^{hu4581/hu4581}; enpp1^sa156/sa156	standard conditions	Days 7-13	axial fin skeleton mineralized, abnormal	Fig. 3
enpp1^{hu4581/hu4581}; enpp1^sa156/sa156	standard conditions	Days 7-13	bone mineralization increased process quality, abnormal	Fig. 3
enpp1^{hu4581/hu4581}; enpp1^sa156/sa156	standard conditions	Days 7-13	vertebra centrum fused with centrum, abnormal	Fig. 3
enpp1^{hu4581/hu4581}; entpd5a^{hu3718/hu3718}	standard conditions	Days 7-13	bone tissue mineralized, normal	Fig. 4
entpd5a^{hu3718/hu3718}	standard conditions	Day 6	bone mineralization disrupted, abnormal	Fig. 1
entpd5a^{hu3718/hu3718}	standard conditions	Day 6	cartilage development process quality, normal	Fig. 1

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
hu3718		Point Mutation	entpd5a
hu4581		Point Mutation	enpp1
hu5310		Point Mutation	entpd5a
hu5939Tg	TgBAC(entpd5a:YFP)	Transgenic Insertion
hu7432Tg	TgBAC(enpp1:enpp1,CMV:YFP)	Transgenic Insertion
sa156		Point Mutation	enpp1
zf132Tg	Tg(Ola.Sp7:NLS-GFP)	Transgenic Insertion

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Human Disease / Model

Sequence Targeting Reagents

Fish

Fish
enpp1^{hu4581/hu4581}
enpp1^{hu4581/hu4581}; enpp1^sa156/sa156
enpp1^{hu4581/hu4581}; entpd5a^{hu3718/hu3718}
entpd5a^{hu3718/hu3718}
entpd5a^{hu3718/hu3718}; zf132Tg
entpd5a^{hu5310/hu5310}
hu5939Tg
hu7432Tg
WT
zf132Tg

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Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
GFP	EFG	GFP
YFP	EFG	YFP

Mapping

Huitema et al., 2012 ZDB-PUB-121220-19 PMID:23236130

Entpd5 is essential for skeletal mineralization and regulates phosphate homeostasis in zebrafish

Huitema et al., 2012

ZDB-PUB-121220-19
PMID:23236130