ArticleBMC developmental biology2016
Revisiting in vivo staining with alizarin red S--a valuable approach to analyse zebrafish skeletal mineralization during development and regeneration.
Article in BMC developmental biology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 62 papers.
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Who cites it
62 citing papers in PubMed, 127 citations in OpenAlex.
- Article
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- Live longitudinal imaging of meningeal cerebrovascular injury and its sequelae in adult zebrafish.bioRxiv : the preprint server for biology · 2025Article
- Dendritic atoh1a+ cells serve as Merkel cell precursors during skin development and regeneration.Development (Cambridge, England) · 2025Article
- [Effect of astragaloside IV on osteogenic differentiation of BMSCs in osteoporotic rats via regulation of miR-21 and inhibition of the Notch signaling pathway].Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences · 2025Article
- Modeling traumatic brain and neural injuries: insights from zebrafish.Frontiers in molecular neuroscience · 2025Review
- Zebrafish Models for Skeletal and Extraskeletal Osteogenesis Imperfecta Features: Unveiling Pathophysiology and Paving the Way for Drug Discovery.Calcified tissue international · 2024Review
- Standardization of bone morphometry and mineral density assessments in zebrafish and other small laboratory fishes using X-ray radiography and micro-computed tomography.Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research · 2024Article
- The dwarf neon rainbowfish Melanotaenia praecox, a small spiny-rayed fish with potential as a new Acanthomorpha model fish: I. Fin ray ontogeny and postembryonic staging.Developmental dynamics : an official publication of the American Association of Anatomists · 2024Article
- Matrix first, minerals later: fine-tuned dietary phosphate increases bone formation in zebrafish.JBMR plus · 2024Article
- Adjusting UV-Vis Spectrum of Alizarin by Insertion of Auxochromes.ChemistryOpen · 2024Article
- Aortic calcification accelerates cardiac dysfunction via inducing apoptosis of cardiomyocytes.International journal of medical sciences · 2024Article
- Improvement of osteogenesis and antibacterial properties of a bioactive glass/gelatin-based scaffold using zoledronic acid and CM11 peptide.Veterinary research forum : an international quarterly journal · 2024Article
- COL11A2 as a candidate gene for vertebral malformations and congenital scoliosis.Human molecular genetics · 2023Article
- In Vitro Screening Studies on Eight Commercial Essential Oils-Derived Compounds to Identify Promising Natural Agents for the Prevention of Osteoporosis.Biomedicines · 2023Article
- mTORC1 signaling pathway regulates tooth repair.International journal of oral science · 2023Article
- Zebrafish Models to Study Ectopic Calcification and Calcium-Associated Pathologies.International journal of molecular sciences · 2023Review
- Article
- Fin ray branching is defined by TRAPProceedings of the National Academy of Sciences of the United States of America · 2022Article
- New insights into detecting alizarin from autofluorescence in marked glass eels.Scientific reports · 2022Article
2 more citing papers are in PubMed but not listed here.
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Authors and funding
6 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundThe correct evaluation of mineralization is fundamental for the study of skeletal development, maintenance, and regeneration. Current methods to visualize mineralized tissue in zebrafish rely on: 1) fixed specimens; 2) radiographic and μCT techniques, that are ultimately limited in resolution; or 3) vital stains with fluorochromes that are indistinguishable from the signal of green fluorescent protein (GFP)-labelled cells. Alizarin compounds, either in the form of alizarin red S (ARS) or alizarin complexone (ALC), have long been used to stain the mineralized skeleton in fixed specimens from all vertebrate groups. Recent works have used ARS vital staining in zebrafish and medaka, yet not based on consistent protocols. There is a fundamental concern on whether ARS vital staining, achieved by adding ARS to the water, can affect bone formation in juvenile and adult zebrafish, as ARS has been shown to inhibit skeletal growth and mineralization in mammals.
resultsHere we present a protocol for vital staining of mineralized structures in zebrafish with a low ARS concentration that does not affect bone mineralization, even after repetitive ARS staining events, as confirmed by careful imaging under fluorescent light. Early and late stages of bone development are equally unaffected by this vital staining protocol. From all tested concentrations, 0.01% ARS yielded correct detection of bone calcium deposits without inducing additional stress to fish.
conclusionsThe proposed ARS vital staining protocol can be combined with GFP fluorescence associated with skeletal tissues and thus represents a powerful tool for in vivo monitoring of mineralized structures. We provide examples from wild type and transgenic GFP-expressing zebrafish, for endoskeletal development and dermal fin ray regeneration.
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