Crtap and p3h1 knock out zebrafish support defective collagen chaperoning as the cause of their osteogenesis imperfecta phenotype

Tonelli, Francesca; Cotti, Silvia; Leoni, Laura; Besio, Roberta; Gioia, Roberta; Marchese, Loredana; Giorgetti, Sofia; Villani, Simona; Gistelinck, Charlotte; Wagener, Raimund; Kobbe, Birgit; Fiedler, Imke A.K.; Larionova, Daria; Busse, Björn; Eyre, David R.; Rossi, Antonio; Witten, P. Eckhard; Forlino, Antonella

doi:10.1016/j.matbio.2020.03.004

Cited by 30 publications

(34 citation statements)

References 56 publications

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“…CRISPR/Cas9 gene editing has been recently used to generate knock-out zebrafish for crtap and p3h1, two genes that are part of a protein complex which is involved in prolyl 3-hydroxylation and proper folding of collagen type I. Loss-of-function mutations in the human ortholog genes cause recessive forms of OI. These zebrafish models faithfully mimic the human disease and support the defective chaperone role of the 3-hydroxylation complex as the primary cause of the skeletal phenotype (17).…”

Section: Reverse Genetic Approach: Morpholino Knockdown and Gene Editingmentioning

confidence: 58%

“…TEM represents a powerful method to analyze ultrastructural features of tissues since it provides much higher magnification and resolution compared to light microscopy, allowing visualization of cellular and matrix structures at a subnanometer scale. For instance, an altered distribution of bone collagen fiber diameter, a frequently described feature in various skeletal pathological conditions, was detected in the crtap and p3h1 knock-out models of OI type VII and VIII by TEM, revealing the crucial role of the collagen post translational modification complex in bone organization (17). TEM was also used to show enlarged endoplasmic reticulum cisterna in these models, reinforcing ER stress as a key element in the OI phenotype and a potential target for new therapeutic approaches (17,226,227).…”

Section: Transmission Electron Microscopy Analysismentioning

confidence: 98%

“…The haploid zebrafish genome has 25 chromosomes containing 1.7 billion base pairs (4). Various forward and reverse genetic approaches have been applied to generate mutant lines that mimic many different human diseases, including skeletal diseases ranging from secondary osteoporosis (OP) to rare disorders such as osteogenesis imperfecta (OI) (12)(13)(14)(15)(16)(17)(18)(19)(20). A major benefit of zebrafish is the simplicity of combining mutant and transgenic lines that express fluorescent reporter proteins under the control of responsive elements for signaling pathways or promoters of cell-type-specific markers.…”

Section: Introductionmentioning

confidence: 99%

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Zebrafish: A Resourceful Vertebrate Model to Investigate Skeletal Disorders

et al. 2020

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Animal models are essential tools for addressing fundamental scientific questions about skeletal diseases and for the development of new therapeutic approaches. Traditionally, mice have been the most common model organism in biomedical research, but their use is hampered by several limitations including complex generation, demanding investigation of early developmental stages, regulatory restrictions on breeding, and high maintenance cost. The zebrafish has been used as an efficient alternative vertebrate model for the study of human skeletal diseases, thanks to its easy genetic manipulation, high fecundity, external fertilization, transparency of rapidly developing embryos, and low maintenance cost. Furthermore, zebrafish share similar skeletal cells and ossification types with mammals. In the last decades, the use of both forward and new reverse genetics techniques has resulted in the generation of many mutant lines carrying skeletal phenotypes associated with human diseases. In addition, transgenic lines expressing fluorescent proteins under bone cell-or pathway-specific promoters enable in vivo imaging of differentiation and signaling at the cellular level. Despite the small size of the zebrafish, many traditional techniques for skeletal phenotyping, such as x-ray and microCT imaging and histological approaches, can be applied using the appropriate equipment and custom protocols. The ability of adult zebrafish to remodel skeletal tissues can be exploited as a unique tool to investigate bone formation and repair. Finally, the permeability of embryos to chemicals dissolved in water, together with the availability of large numbers of small-sized animals makes zebrafish a perfect model for high-throughput bone anabolic drug screening. This review aims to discuss the techniques that make zebrafish a powerful model to investigate the molecular and physiological basis of skeletal disorders.

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Section: Reverse Genetic Approach: Morpholino Knockdown and Gene Editingmentioning

confidence: 58%

Section: Transmission Electron Microscopy Analysismentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

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Zebrafish: A Resourceful Vertebrate Model to Investigate Skeletal Disorders

et al. 2020

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“…Diameters were measured on fully transversely sectioned fibres using ImageJ software (NIH, Bethesda, MD, USA). Analysis of collagen fibres was based on previously established protocol [ 36 ], the number of fibres analysed exceeds the established one.…”

Section: Methodsmentioning

confidence: 99%

“…Danio rerio (zebrafish) is an established model for the study of bone formation, given that basic bone cell differentiation pathways and ossification processes have been conserved across vertebrates [ 35 ]. This study examines the post-cranial skeleton in zebrafish, given that the vertebral column is the most studied anatomical structure in biomedical research and in aquaculture [ 36 , 37 ]. Likewise, fin rays are popular for the study of bone regeneration and fish health [ 38 , 39 ].…”

Section: Introductionmentioning

confidence: 99%

More Bone with Less Minerals? The Effects of Dietary Phosphorus on the Post-Cranial Skeleton in Zebrafish

Cotti

Huysseune

Koppe³

et al. 2020

IJMS

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Dietary phosphorus (P) is essential for bone mineralisation in vertebrates. P deficiency can cause growth retardation, osteomalacia and bone deformities, both in teleosts and in mammals. Conversely, excess P supply can trigger soft tissue calcification and bone hypermineralisation. This study uses a wide range of complementary techniques (X-rays, histology, TEM, synchrotron X-ray tomographic microscopy, nanoindentation) to describe in detail the effects of dietary P on the zebrafish skeleton, after two months of administering three different diets: 0.5% (low P, LP), 1.0% (regular P, RP), and 1.5% (high P, HP) total P content. LP zebrafish display growth retardation and hypomineralised bones, albeit without deformities. LP zebrafish increase production of non-mineralised bone matrix, and osteoblasts have enlarged endoplasmic reticulum cisternae, indicative for increased collagen synthesis. The HP diet promotes growth, high mineralisation, and stiffness but causes vertebral centra fusions. Structure and arrangement of bone matrix collagen fibres are not influenced by dietary P in all three groups. In conclusion, low dietary P content stimulates the formation of non-mineralised bone without inducing malformations. This indicates that bone formation and mineralisation are uncoupled. In contrast, high dietary P content promotes mineralisation and vertebral body fusions. This new zebrafish model is a useful tool to understand the mechanisms underlying osteomalacia and abnormal mineralisation, due to underlying variations in dietary P levels.

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A new case of osteogenesis imperfecta type VIII and retinal detachment

Souza

Nunes

Magalhães

et al. 2020

American J of Med Genetics Pt A

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Osteogenesis imperfecta (OI) type VIII (OMIM: 610915) is a rare autosomal recessive disorder characterized by white sclerae, severe growth deficiency, and bone fragility. This condition results from pathogenic variants of P3H1, a gene that codes for P3H1, an important protein involved in the prolyl‐3‐hydroxylation complex required for collagen type I folding. Here, we described a woman with OI type VIII due to a homozygous mutation of c.1914+1G>C (NM_001243246.1) in P3H1 and retinal detachment. We compared our case to five severe OI and retinal detachment cases reported in the literature. The only case previously reported with a molecular diagnosis had a similar mutation in P3H1 c.1914+1G>A and a giant retinal detachment. We suggest that individuals with OI type VIII should be submitted to careful fundoscopic examination.

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Crtap and p3h1 knock out zebrafish support defective collagen chaperoning as the cause of their osteogenesis imperfecta phenotype

Cited by 30 publications

References 56 publications

Zebrafish: A Resourceful Vertebrate Model to Investigate Skeletal Disorders

Zebrafish: A Resourceful Vertebrate Model to Investigate Skeletal Disorders

More Bone with Less Minerals? The Effects of Dietary Phosphorus on the Post-Cranial Skeleton in Zebrafish

A new case of osteogenesis imperfecta type VIII and retinal detachment

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