zOPT: an open source optical projection tomography system and methods for rapid 3D zebrafish imaging

Zhang, Hanqing; Waldmann, Laura; Manuel, Remy; Boije, Henrik; Haitina, Tatjana; Allalou, Amin

doi:10.1364/boe.393519

Cited by 15 publications

(13 citation statements)

References 33 publications

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“…A custom-built Optical Projection Tomography (OPT) system was used for imaging of the zebrafish embryos at 5 dpf that were euthanised with an overdose of MS-222 (300mg/L), fixed and stained with alcian blue [ 35 , 36 ]. The OPT system, reconstruction algorithms, and alignment workflow were based on the previously described method [ 37 ].…”

Section: Methodsmentioning

confidence: 99%

The broad role of Nkx3.2 in the development of the zebrafish axial skeleton

et al. 2021

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The transcription factor Nkx3.2 (Bapx1) is an important chondrocyte maturation inhibitor. Previous Nkx3.2 knockdown and overexpression studies in non-mammalian gnathostomes have focused on its role in primary jaw joint development, while the function of this gene in broader skeletal development is not fully described. We generated a mutant allele of nkx3.2 in zebrafish with CRISPR/Cas9 and applied a range of techniques to characterize skeletal phenotypes at developmental stages from larva to adult, revealing loss of the jaw joint, fusions in bones of the occiput, morphological changes in the Weberian apparatus, and the loss or deformation of bony elements derived from basiventral cartilages of the vertebrae. Axial phenotypes are reminiscent of Nkx3.2 knockout in mammals, suggesting that the function of this gene in axial skeletal development is ancestral to osteichthyans. Our results highlight the broad role of nkx3.2 in zebrafish skeletal development and its context-specific functions in different skeletal elements.

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Section: Methodsmentioning

confidence: 99%

The broad role of Nkx3.2 in the development of the zebrafish axial skeleton

et al. 2021

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“…A custom-built Optical Projection Tomography (OPT) system was used for imaging of the zebrafish embryos fixed at 5 dpf and stained with alcian blue (Sharpe et al, 2002;Zhang et al, 2020). The OPT system, reconstruction algorithms, and alignment workflow were based on the previously described method (Allalou et al, 2017).…”

Section: Optical Projection Tomographymentioning

confidence: 99%

The Broad Role of Nkx3.2 in the Development of the Zebrafish Axial Skeleton

Waldmann

Leyhr

Zhang

et al. 2020

Preprint

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The transcription factor Nkx3.2 (Bapx1) is an important chondrocyte maturation inhibitor. Previous Nkx3.2 knock-down and overexpression studies in non-mammalian gnathostomes have focused on its role in primary jaw joint development, while little is known about the function of this gene in broader skeletal development. We generated CRISPR/Cas9 knockout of nkx3.2 in zebrafish and applied a range of techniques to characterize skeletal phenotypes at developmental stages from larva to adult, revealing fusions in bones of the occiput, the loss or deformation of bony elements derived from basiventral cartilages of the vertebrae, and an increased length of the proximal radials of the dorsal and anal fins. These phenotypes are reminiscent of Nkx3.2 knockout phenotypes in mammals, suggesting that the function of this gene in axial skeletal development is ancestral to osteichthyans. Our results highlight the broad role of nkx3.2 in zebrafish skeletal development and its context-specific functions in different skeletal elements.

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“…A custom-built Optical Projection Tomography (OPT) system was used for imaging of the zebrafish embryos fixed at 9 dpf and stained cartilage and bone (Sharpe et al, 2002; Zhang et al, 2020). The OPT system, reconstruction algorithms, and alignment workflow were based on the previously described method (Allalou et al, 2017).…”

Section: Methodsmentioning

confidence: 99%

The role of Gdf5 in the development of the zebrafish fin endoskeleton

Waldmann

Leyhr

Zhang

et al. 2021

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BackgroundThe development of the vertebrate limb skeleton requires a complex interaction of multiple factors to facilitate correct shaping and positioning of bones and joints. Growth and differentiation factor 5 (Gdf5), a member of the transforming growth factor-beta family (TGF-β) is involved in patterning appendicular skeletal elements including joints. Expression of gdf5 in zebrafish has been detected within the first pharyngeal arch jaw joint, fin mesenchyme condensations and segmentation zones in median fins, however little is known about the functional role of Gdf5 outside of Amniota.ResultsWe generated CRISPR/Cas9 knockout of gdf5 in zebrafish and analysed the resulting phenotype at different developmental stages. Homozygous gdf5 mutant zebrafish displayed changes in segmentation of the endoskeletal disc and, in consequence, loss of posterior radials in the pectoral fins. Mutant fish also displayed affected organisation and length of skeletal elements in the median fins, however joint formation and mineralisation process seemed unaffected.ConclusionsOur study demonstrates the importance of Gdf5 for the paired and median fin endoskeleton development in zebrafish and reveals that the severity of the effect increases from anterior to posterior side of the elements. Our findings are consistent with phenotypes observed in human and mouse appendicular skeleton in response to Gdf5 knockout, suggesting a broadly conserved role for Gdf5 in Osteichthyes.

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zOPT: an open source optical projection tomography system and methods for rapid 3D zebrafish imaging

Cited by 15 publications

References 33 publications

The broad role of Nkx3.2 in the development of the zebrafish axial skeleton

The broad role of Nkx3.2 in the development of the zebrafish axial skeleton

The Broad Role of Nkx3.2 in the Development of the Zebrafish Axial Skeleton

The role of Gdf5 in the development of the zebrafish fin endoskeleton

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