A rapid and nondestructive protocol for whole-mount bone staining of small fish and Xenopus

Sakata‐Haga, Hiromi; Uchishiba, Maimi; Shimada, Hiroki; Tsukada, Tsuyoshi; Mitani, Mayumi; Arikawa, Tomohiro; Shoji, Hiromichi; Hatta, Toshihisa

doi:10.1038/s41598-018-25836-4

Cited by 35 publications

(34 citation statements)

References 9 publications

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“…We also evaluated the bone deformation caused by Cd pathotoxicity, similar to human cases reported as osteomalacia in postmenopausal women upon chronic Cd exposure (Huff et al ; Lane et al ; Aoshima ). We analyzed the whole‐mount bone staining with alizarin red S (Sakata‐Haga et al ). We observed normal bone morphology at 0 ppm Cd in 3‐month‐old and 12‐month‐old female and male medaka (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Extracellular vesicles from human bone marrow mesenchymal stem cells repair organ damage caused by cadmium poisoning in a medaka model

et al. 2019

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Treatment modalities for kidney disease caused by long‐term exposure to heavy metals, such as cadmium (Cd), are limited. Often, chronic, long‐term environmental exposure to heavy metal is not recognized in the early stages; therefore, chelation therapy is not an effective option. Extracellular vesicles (EVs) derived from stem cells have been demonstrated to reduce disease pathology in both acute and chronic kidney disease models. To test the ability of EVs derived from human bone marrow mesenchymal stem cells (hBM‐MSCs) to treat Cd damage, we generated a Cd‐exposed medaka model. This model develops heavy metal‐induced cell damage in various organs and tissues, and shows decreased overall survival. Intravenous injection of highly purified EVs from hBM‐MSCs repaired the damage to apical and basolateral membranes and mitochondria of kidney proximal tubules, glomerular podocytes, bone deformation, and improved survival. Our system also serves as a model with which to study age‐ and sex‐dependent cell injuries of organs caused by various agents and diseases. The beneficial effects of EVs on the tissue repair process, as shown in our novel Cd‐exposed medaka model, may open new broad avenues for interventional strategies.

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Section: Resultsmentioning

confidence: 99%

“…Whole‐mount bone staining of medaka was performed as previously described, with the modifications described below (Sakata‐Haga et al ). Tricaine‐S‐anesthetized medaka were immersed in freshly prepared fixative (5% Formalin/5% Triton X‐100/1% KOH) (HT501128, T8787, 221473, Sigma‐Aldrich) for 24 h at 42°C with rotation.…”

Section: Methodsmentioning

confidence: 99%

Extracellular vesicles from human bone marrow mesenchymal stem cells repair organ damage caused by cadmium poisoning in a medaka model

et al. 2019

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“…Many different protocols exist for ARS staining of mineralized tissues, however the main steps are based on (i) removing the pigmentation of the tissue with a bleaching solution (basic pH), (ii) neutralization of depigmentation, (iii) staining the animal with ARS, and (iv) clearing the animal of excess stain ( 244 ). The ARS molecule is a dihydroxyanthraquinone, likely binding the Ca 2+ on the hydroxyapatite surface to form either a salt or a chelate form ( 245 ), thus it specifically stains mineralized tissue.…”

Section: Bone Histology: From Whole Mount To Sectionsmentioning

confidence: 99%

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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“…Although harsh and not applicable to perform immunostaining or XFP visualization, these techniques are still in use, even though the new applications are being proposed, such as observation of ectopic calcium deposits in dystrophin‐deficient mdx mouse muscles generated upon dietary phosphorus overload . Recently, KOH‐based clearing approach was optimized by Sakata‐Haga and colleagues, who described the first fast (3 days long), and nondestructive variation of this whole‐mount protocol for the efficient clearing and visualization of bone in zebrafish and Xenopus frog.…”

Section: Application Of Toc For Particular Peripheral Organsmentioning

confidence: 99%

Advances in Ex Situ Tissue Optical Clearing

Matryba

Kaczmarek

Gołąb

2019

Laser & Photonics Reviews

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Examination of whole organs with subcellular resolution in health, disease, and during development is necessary to decipher their biological complexity. However, until recently, this has been virtually impossible due to the natural opacity of organ tissue. Recent progress in tissue optical clearing (TOC) has overcome this limitation by turning organs into transparent, light-permitting specimens. At least 20 original TOC methods have been developed in less than a decade, which were followed by hundreds of attempts that were aimed at their optimization and practical application. The majority of proof-of-concept studies have focused on the brain. However, it is apparent that TOC might be equally valuable when applied to peripheral organs or even the whole body. The progress in TOC for peripheral organs is delineated in an organ-by-organ fashion and whole-body clearing approaches are discussed. Additionally, physical and optical approaches for TOC affecting the optical properties of the samples and image quality are discussed to explore their advantages, limitations, and future possibilities.

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A rapid and nondestructive protocol for whole-mount bone staining of small fish and Xenopus

Cited by 35 publications

References 9 publications

Extracellular vesicles from human bone marrow mesenchymal stem cells repair organ damage caused by cadmium poisoning in a medaka model

Extracellular vesicles from human bone marrow mesenchymal stem cells repair organ damage caused by cadmium poisoning in a medaka model

Zebrafish: A Resourceful Vertebrate Model to Investigate Skeletal Disorders

Advances in Ex Situ Tissue Optical Clearing

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