Assessing the ability of zebrafish scales to contribute to the short-term homeostatic regulation of [Ca2+] in the extracellular fluid during calcemic challenges

  • Jacky T. Hung
  • Sarah E. Webb
  • Carla Palumbo
  • Agnieszka M. Lesniak
  • Alan M. Shipley
  • Alessandro Rubinacci
  • Joseph G. Kunkel
  • Andrew L. MillerEmail author
Original Article Biology


The elasmoid scales of fish represent a significant internal reservoir of calcium ions (Ca2+), but little is known about the contribution of these scales to the short-term regulation of Ca2+ homeostasis in the extracellular fluid (ECF). This gap in our knowledge is partly due to the technical challenges involved in measuring small Ca2+ fluxes around the scales of live fish in real time. Here, we describe a technique for exfoliating, mounting, and culturing intact living zebrafish Danio rerio scales, then subjecting them to examination using an extracellular, non-invasive, surface-scanning ion-selective electrode technique (SIET). In a Ca2+-sensitive configuration, the SIET can resolve Ca2+ flux values in the low-to-sub picomole/square centimeter/second range, with a spatial resolution of approximately 5 μm. We quantified the Ca2+ fluxes into and out of scales under different extracellular calcemic challenges set to mimic a variety of Ca2+ concentrations in the ECF and showed that the results were similar to those previously reported from isolated mouse metatarsal bone. Our new data extend our current understanding of the role played by fish scales in the short-term homeostatic regulation of Ca2+ concentration in the ECF. They also support the suggestion that scales might provide an inexpensive and complementary model for studying the fundamentals of bone-mediated homeostatic Ca2+ regulation and the diseases that result from its dysregulation.


Extracellular fluid Ca2+ homeostasis Scanning ion-selective electrode technique Zebrafish scales 



This project was supported by the Hong Kong Research Grants Council General Research Fund awards: 16101714 and 16100115. We also acknowledge funding from the Hong Kong Innovation and Technology Commission (ITCPD/17-9). This work was made possible by equipment and support generously provided by Mr. Chris Shipley of Applicable Electronics, LLC., New Haven, CT, USA, and software kindly donated by Mr. Eric Karplus of Science Wares Inc., Falmouth, MA, USA. Initial experiments were carried out by JTH at the MBL (Woods Hole) during the Croucher Foundation-funded JUSTL Program.

Author contributions

JTH was involved in developing the methodology, conducting the research, and preparing the manuscript. SEW helped to supervise the project and was involved with conducting the research, verifying the results, and preparing the manuscript. CP was involved in conducting some of the research and preparing the manuscript. AML was involved in conducting some of the research. AMS helped design the methodology, provided the instrumentation, and helped with student training. AR helped to formulate the research goals and was involved in preparing the manuscript. JGK helped to formulate the research goals and provided advice regarding the methodology and data analysis. ALM formulated the overarching research goals, supervised the project, provided the resources for conducting the project, and prepared the manuscript. All the authors helped review the manuscript.


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Copyright information

© Japanese Society of Fisheries Science 2019

Authors and Affiliations

  1. 1.Division of Life Science and State Key Laboratory for Molecular NeuroscienceThe Hong Kong University of Science and Technology (HKUST)Hong KongChina
  2. 2.Section of Human Morphology, Department of Biomedical, Metabolic and Neural SciencesUniversity of Modena and Reggio EmiliaModenaItaly
  3. 3.Applicable Electronics, LLCNew HavenUSA
  4. 4.Bone Metabolism UnitScientific Institute San RaffaeleMilanItaly
  5. 5.Pickus Center for Biomedical ResearchUniversity of New EnglandBiddefordUSA

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