Journal of Integrative Natural Science (통합자연과학논문집)

The Basic Science Institute Chosun University (조선대학교 기초과학연구원)
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A Practical Protocol of Zebrafish Heart Rate Measurement for High School Students

  • Received : 2021.11.09
  • Accepted : 2021.12.29
  • Published : 2021.12.31
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Abstract

To study the effects of hormones and neurotransmitters, zebrafish (Danio rerio) are a great substitute for water fleas (Daphnia). The zebrafish is an ideal vertebrate model because it has a transparent embryonic stage. It is easy to get consistent heart rate measurements in embryonic zebrafish when treating them with hormones and neurotransmitters. To observe the heart rate, two to three embryonic zebrafish are anesthetized with MS-222 and then transferred to a glass slide specifically designed for heart observation and easy application of various chemicals. After the heartbeats are counted for 2 minutes, apply either 100 µM epinephrine or 100 µM acetylcholine to the zebrafish. Wait 5, 10, and 20 minutes and count the heartbeats at each time point. All procedures are repeated three times. The final results are averaged and analyzed by using statistical methods. The above method which we have developed is practical enough for high school students to measure the heart rate in zebrafish under various conditions and to analyze the data set.

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Acknowledgement

The study was supported by a research fund from Chosun University, 2020. We would also like to thank Dr. Eleanor Thorne for her careful reading of the manuscript and for great comments to improve and clarify the manuscript.

References

  1. Corotto, F., D. Ceballos, A. Lee, L. Vinson, "Making the Most of the Daphnia Heart Rate Lab: Optimizing the Use of Ethanol, Nicotine & Caffeine", Am Biol Teach, Vol. 72, pp. 176-179, 2010. https://doi.org/10.1525/abt.2010.72.3.9
  2. Bekker, J. M., B. J. Krijgsman, "Physiological Investigations into the Heart Function of Daphnia", J Physiol, Vol. 115, pp. 249-257, 1951. https://doi.org/10.1113/jphysiol.1951.sp004669
  3. Schwerte, T., C. Prem, A. Mairosl, B. Pelster, "Development of the Sympatho-Vagal Balance in the Cardiovascular System in Zebrafish (Danio Rerio) Characterized by Power Spectrum and Classical Signal Analysis", J Exp Biol, Vol. 209, pp. 1093-1100, 2006. https://doi.org/10.1242/jeb.02117
  4. Meyers, J. R., "Zebrafish: Development of a Vertebrate Model Organism", Current Protocols Essential Laboratory Techniques, Vol. 16, pp. e19, 2018. https://doi.org/10.1002/cpet.19
  5. Nikam, V. S., D. Singh, R. Takawale, M. R. Ghante, "Zebrafish: An Emerging Whole-Organism Screening Tool in Safety Pharmacology", Indian journal of pharmacology, Vol. 52, pp. 505-513, 2020. https://doi.org/10.4103/ijp.IJP_482_19
  6. Poon, K. L., T. Brand, "The Zebrafish Model System in Cardiovascular Research: A Tiny Fish with Mighty Prospects", Global Cardiology Science and Practice, Vol. 2013, pp. 2013.
  7. 조정훈, 김선영, "제브라피시를 활용한 동물의 생식과 발생 및 순환 관련 탐구 모듈 개발 가능성 탐색", 생물교육(구 생물교육학회지), Vol. 41, pp. 392-404, 2013.
  8. Craig, M. P., S. D. Gilday, J. R. Hove, "Dose-Dependent Effects of Chemical Immobilization on the Heart Rate of Embryonic Zebrafish", Lab Animal, Vol. 35, pp. 41-47, 2006. https://doi.org/10.1038/laban1006-41
  9. Parish, H. G., J. R. Morton, J. C. Brown, "A Systematic Review of Epinephrine Stability and Sterility with Storage in a Syringe", Allergy, Asthma & Clinical Immunology, Vol. 15, pp. 7, 2019. https://doi.org/10.1186/s13223-019-0324-7
  10. Saleheen, A., B. M. Campbell, R. A. Prosser, C. A. Baker, "Estimation of Available Epinephrine Dose in Expired and Discolored Autoinjectors Via Quantitative Smartphone Imaging", Anal Bioanal Chem, Vol. 412, pp. 2785-2793, 2020. https://doi.org/10.1007/s00216-020-02505-y
  11. Trautner, E. M., T. R. Bradley, "The Early Stages of the Oxidation of Adrenaline in Dilute Solution", Aust J Sci Res B, Vol. 4, pp. 303-343, 1951. https://doi.org/10.1071/BI9510303
  12. Sletten, D. M., K. K. Nickander, P. A. Low, "Stability of Acetylcholine Chloride Solution in Autonomic Testing", J Neurol Sci, Vol. 234, pp. 1-3, 2005. https://doi.org/10.1016/j.jns.2005.02.007