Zebrafish as a unique model system in bone research: the power of genetics and<i>in vivo</i>imaging

Spoorendonk, Kirsten M.; Hammond, Chrissy L.; Huitema, Leonie F. A.; Vanoevelen, Jo; Schulte‐Merker, Stefan

doi:10.1111/j.1439-0426.2010.01409.x

Cited by 71 publications

(59 citation statements)

References 39 publications

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“…In a forward genetic screen in zebrafish (14), we uncovered 14 mutant lines out of 429 families screened. One mutant, no bone (nob hu3718 ), completely lacked a mineralized skeleton ( Fig.…”

Section: Resultsmentioning

confidence: 99%

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

Huitema

Apschner

Logister

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

100

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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-offunction 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.

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Section: Resultsmentioning

confidence: 99%

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

Huitema

Apschner

Logister

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

100

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“…A forward-genetic screen was performed using Alizarin Red staining as described previously (Spoorendonk et al, 2010). The mutation in abcc6a was detected using whole-genome sequencing, as reported elsewhere ( Mackay and Schulte-Merker, 2014).…”

Section: Identification Of the Gräte Mutationmentioning

confidence: 99%

Vitamin K reduces hypermineralisation in zebrafish models of PXE and GACI

2015

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The mineralisation disorder pseudoxanthoma elasticum (PXE) is associated with mutations in the transporter protein ABCC6. Patients with PXE suffer from calcified lesions in the skin, eyes and vasculature, and PXE is related to a more severe vascular calcification syndrome called generalised arterial calcification of infancy (GACI). Mutations in ABCC6 are linked to reduced levels of circulating vitamin K. Here, we describe a mutation in the zebrafish (Danio rerio) orthologue abcc6a, which results in extensive hypermineralisation of the axial skeleton. Administration of vitamin K to embryos was sufficient to restore normal levels of mineralisation. Vitamin K also reduced ectopic mineralisation in a zebrafish model of GACI, and warfarin exacerbated the mineralisation phenotype in both mutant lines. These data suggest that vitamin K could be a beneficial treatment for human patients with PXE or GACI. Additionally, we found that abcc6a is strongly expressed at the site of mineralisation rather than the liver, as it is in mammals, which has significant implications for our understanding of the function of ABCC6.

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“…The zebrafish is also a very attractive model for studying the mechanisms underlying bone formation, 29,30 because the key regulators of bone formation are highly conserved between mammals and teleosts, and the corresponding orthologs share significant sequence similarities and an overlap in expression patterns when compared with mammals. [31][32][33] Molecules essential to promoting osteoclastogenesis 34 include (1) macrophage colony-stimulating factor (M-CSF), (2) receptor for activation of tumor necrosis factor kappa B (TNF-κB or receptor for necrosis factor [RANK]), and (3) RANK ligand (RANK-L, OPGL, or TRANCE).…”

mentioning

confidence: 99%

Mesoporous silica nanoparticles as a compound delivery system in zebrafish embryos

Sharif¹,

Porta²,

Meijer³

et al. 2012

IJN

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Silica nanoparticles can be efficiently employed as carriers for therapeutic drugs in vitro. Here, we use zebrafish embryos as a model organism to see whether mesoporous silica nanoparticles (MSNPs) can be incorporated to deliver compounds in vivo. We injected 35-40 nL (10 mg/mL) of custom-synthesized, fluorescently-tagged 200 nm MSNPs into the left flank, behind the yolk sac extension, of 2-day-old zebrafish embryos. We tracked the distribution and translocation of the MSNPs using confocal laser scanning microscopy. Some of the particles remained localized at the injection site, whereas others entered the bloodstream and were carried around the body. Embryo development and survival were not significantly affected by MSNP injection. Acridine orange staining revealed that MSNP injections did not induce significant cell death. We also studied cellular immune responses by means of lysC::DsRED2 transgenic embryos. MSNP-injected embryos showed infiltration of the injection site with neutrophils, similar to controls injected with buffer only. In the same embryos, counterstaining with L-plastin antibody for leukocytes revealed the same amount of cellular infiltration of the injection site in embryos injected with MSNPs or with buffer only. Next, we used MSNPs to deliver two recombinant cytokines (macrophage colony-stimulating factor and receptor for necrosis factor ligand) to zebrafish embryos. These proteins are known to activate cells involved in bone remodeling, and this can be detected with the marker tartrate-resistant acid phosphatase. Coinjection of these proteins loaded onto MSNPs produced a significant increase in the number of tartrate-resistant acid phosphatase-positive cells after 2-3 days of injection. Our results show that MSNPs can be used to deliver bioactive compounds into zebrafish larvae without producing higher mortality or gross evidence of teratogenicity.

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Zebrafish as a unique model system in bone research: the power of genetics andin vivoimaging

Cited by 71 publications

References 39 publications

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

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

Vitamin K reduces hypermineralisation in zebrafish models of PXE and GACI

Mesoporous silica nanoparticles as a compound delivery system in zebrafish embryos

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