Zebrafish (Danio rerio) as a Viable Model to Study the Blood-Brain Barrier

  • Tianzhi Yang
  • Shuhua BaiEmail author
Part of the Neuromethods book series (NM, volume 142)


As the blood-brain barrier (BBB) is essential for maintaining brain homeostasis and protecting the brain from exogenous substances, impermeability of the BBB is a major obstacle for drug delivery into the brain. Under pathological conditions, the integrity of the BBB is susceptible to disruption and can be broken down in severe brain diseases. Therefore, the understanding of intrinsic complexity as well as modulation of the BBB is critical to discover potential therapeutics for the treatment of brain diseases. Zebrafish (Danio rerio) have emerged as a suitable animal model in studying pathology of diseases and screening leading compounds in the drug development and discovery because of their highly conserved nature in both genetics and cell biology as higher vertebrates. Importantly, due to their small body size, ease of care, rapid development, and transparency in the early embryo stage, zebrafish allow researchers to study the BBB and carry out high-throughput screening of potential therapeutics with cost-effectiveness. We thus aim to provide a technical overview of the procedures that can be used to analyze BBB integrity and functionality in zebrafish. Low permeability and strong tight junction-based BBB in zebrafish are very similar to those of higher vertebrates. Zebrafish could be an excellent experimental model organism for studying the development and maintenance of the BBB, defining disease pathway, and discovering specific and powerful therapies for the treatment of brain diseases.

Key words

Zebrafish Blood-brain barrier Permeability Efflux Drug delivery Microinjection 


  1. 1.
    Wilhelm I, Fazakas C, Krizbai IA (2011) In vitro models of the blood-brain barrier. Acta Neurobiol Exp (Wars) 71:113–128Google Scholar
  2. 2.
    De Rosa G, Salzano G, Caraglia M, Abbruzzese A (2012) Nanotechnologies: a strategy to overcome blood-brain barrier. Curr Drug Metab 13:61–69CrossRefGoogle Scholar
  3. 3.
    Pardridge WM (2005) The blood-brain barrier: bottleneck in brain drug development. NeuroRx 2:3–14. Scholar
  4. 4.
    Pardridge WM (2007) Blood-brain barrier delivery. Drug Discov Today 12:54–61. Scholar
  5. 5.
    Nicolazzo JA, Charman SA, Charman WN (2006) Methods to assess drug permeability across the blood-brain barrier. J Pharm Pharmacol 58:281–293. Scholar
  6. 6.
    Reichel A, Begley DJ, Abbott NJ (2003) An overview of in vitro techniques for blood-brain barrier studies. Methods Mol Med 89:307–324. Scholar
  7. 7.
    Davis TP, Abbruscato TJ, Egleton RD (2015) Peptides at the blood brain barrier: knowing me knowing you. Peptides 72:50–56. Scholar
  8. 8.
    Keaney J, Campbell M (2015) The dynamic blood-brain barrier. FEBS J 282:4067–4079. Scholar
  9. 9.
    Haseloff RF, Dithmer S, Winkler L, Wolburg H, Blasig IE (2015) Transmembrane proteins of the tight junctions at the blood-brain barrier: structural and functional aspects. Semin Cell Dev Biol 38:16–25. Scholar
  10. 10.
    Kniesel U, Wolburg H (2000) Tight junctions of the blood-brain barrier. Cell Mol Neurobiol 20:57–76CrossRefGoogle Scholar
  11. 11.
    Mahar Doan KM et al (2002) Passive permeability and P-glycoprotein-mediated efflux differentiate central nervous system (CNS) and non-CNS marketed drugs. J Pharmacol Exp Ther 303:1029–1037. Scholar
  12. 12.
    Mahringer A, Ott M, Reimold I, Reichel V, Fricker G (2011) The ABC of the blood-brain barrier - regulation of drug efflux pumps. Curr Pharm Des 17:2762–2770CrossRefGoogle Scholar
  13. 13.
    Rubinstein AL (2003) Zebrafish: from disease modeling to drug discovery. Curr Opin Drug Discov Devel 6:218–223PubMedGoogle Scholar
  14. 14.
    Lieschke GJ, Currie PD (2007) Animal models of human disease: zebrafish swim into view. Nat Rev Genet 8:353–367. Scholar
  15. 15.
    Umans RA, Taylor MR (2012) Zebrafish as a model to study drug transporters at the blood-brain barrier. Clin Pharmacol Ther 92:567–570. Scholar
  16. 16.
    Xie J, Farage E, Sugimoto M, Anand-Apte B (2010) A novel transgenic zebrafish model for blood-brain and blood-retinal barrier development. BMC Dev Biol 10:76. Scholar
  17. 17.
    Santoriello C, Zon LI (2012) Hooked! Modeling human disease in zebrafish. J Clin Invest 122:2337–2343. Scholar
  18. 18.
    Jeong JY et al (2008) Functional and developmental analysis of the blood-brain barrier in zebrafish. Brain Res Bull 75:619–628. Scholar
  19. 19.
    Chakraborty C, Hsu CH, Wen ZH, Lin CS, Agoramoorthy G (2009) Zebrafish: a complete animal model for in vivo drug discovery and development. Curr Drug Metab 10:116–124CrossRefGoogle Scholar
  20. 20.
    Karlsson J, von Hofsten J, Olsson PE (2001) Generating transparent zebrafish: a refined method to improve detection of gene expression during embryonic development. Mar Biotechnol (NY) 3:522–527. Scholar
  21. 21.
    Avdesh A et al (2012) Regular care and maintenance of a zebrafish (Danio rerio) laboratory: an introduction. J Vis Exp e4196.
  22. 22.
    Sive HL, Grainger RM, Harland RM (2010) Calibration of the injection volume for microinjection of Xenopus oocytes and embryos. Cold Spring Harb Protoc 2010:pdb prot5537. Scholar
  23. 23.
    Eliceiri BP, Gonzalez AM, Baird A (2011) Zebrafish model of the blood-brain barrier: morphological and permeability studies. Methods Mol Biol 686:371–378. Scholar
  24. 24.
    Yang T et al (2015) Exosome delivered anticancer drugs across the blood-brain barrier for brain cancer therapy in Danio rerio. Pharm Res 32:2003–2014. Scholar
  25. 25.
    Yang T et al (2017) Delivery of small interfering RNA to inhibit vascular endothelial growth factor in zebrafish using natural brain endothelia cell-secreted exosome nanovesicles for the treatment of brain cancer. AAPS J 19:475–486. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Basic Pharmaceutical Sciences, School of PharmacyHusson UniversityBangorUSA

Personalised recommendations