Effect of Estradiol on Gamete Development and Behavior in Zebrafish Model

Mohammad Mizwaruddin Sidek, Suzanah Abd Rahman


Xenobiotic substance released in the environment is a concern among the public at large. The example of this xenobiotic release into the environment is xenoestrogens. Xenoestrogens have the capability to bind to the estrogen receptors in the body even at low affinity. Food, pesticides and contraceptive pills are known sources of xenoestrogens. In this study, acute toxicity test was conducted to evaluate toxicity of synthetic estrogen such as estradiol to the embryo-larvae of zebrafish model. Morphological changes in the embryo-larvae of zebrafish were also observed. The parameters that were evaluated in acute toxicity study were half lethal concentration (LC50) and few apical endpoints such as coagulation of embryos, development of pericardial edema, and eyes size. Toxicity effect of the compound was evaluated in term of behavior activity of the larvae. Results showed that certain concentration of estradiol caused toxic effects to the embryo-larvae of the zebrafish (p<0.05). The examples of toxic effects that were observed from this test were development of pericardial edema and small eyes in high concentration groups of estradiol used (1700, 850, and 425 mg/L), p<0.05. In terms of behavioral activity, the larvae were greatly affected by the estradiol in which the high group concentrations of 850 and 425 mg/L resulted in the inactivity of the larvae compared with the negative control group, p<0.05. In conclusion, estradiol has effects on mortality rate as indicated by the half lethal concentration (LC50), morphological changes and behavior activity of the embryo-larvae of zebrafish.


Xenobiotic, estradiol, zebrafish, toxicity, development, behaviour.

Full Text:



Ososki , A . L. , & Kennelly , E . J. Phytoestrogens: a review of the present state of research. Phytotherapy Research. 2003; 17(8): 845-869.

Shuster, L. T. , Rhodes , D . J. , Gostout , B. S. , Grossardt , B . R ., & Rocca, W. A. Premature menopause or early menopause: long-term health consequences. Maturitas. 2010; 65(2): 161-166.

Lai, K. M . , Scrimshaw, M. D. , & Lester, J. N. The effects of natural and synthetic steroid estrogens in relation to their environmental occurrence. Critical reviews in toxicology. 2002; 32(2): 113-132.

Sonnenschein , C . , & Soto, A. M. An updated review of environmental estrogen and androgen mimics and antagonists. The Journal of steroid biochemistry and molecular biology. 1998; 65(1): 143-150.

Grady, D., Gebretsadik, T., Kerlikowske, K., Ernster, V., & Petitti, D. Hormone replacement therapy and endometrial cancer risk: a metaanalysis. Obstetrics & Gynecology. 1995; 85(2): 304-313.

Million Women Study Collaborators. Breast cancer and hormone - replacement therapy in the Million Women Study. The Lancet. 2003; 362(9382): 419-427.

Scott, G . R. , & Sloman , K. A. The effects of environmental pollutants on complex fish behavior: integrating behavioral and physiological indicators of toxicity. Aquatic toxicology. 2004; 68(4): 369-392.

Scholz, S., Fischer, S., Gündel, U., Küster, E., Luckenbach, T., & Voelker, D. The zebrafish embryo model in environmental risk assessment—applications beyond acute toxicity testing. Environmental Science and Pollution Research. 2008; 15(5): 394-404.

Kimmel, C. B., Ballard, W. W., Kimmel, S. R., Ullmann, B., & Schilling, T. F. Stages of embryonic development of the zebrafish. Developmental dynamics. 1995; 203(3): 253-310.

Porseryd, T., Volkova, K., Reyhanian Caspillo, N., Källman, T., Dinnetz, P., & Porsh Hällström, I. Persistent effects of developmental exposure to 17αethinylestradiol on the zebrafish (Danio rerio) brain transcriptome and behavior. Frontiers in behavioral neuroscience. 2017; 11: 69.

Hao, R., Bondesson, M., Singh, A. V., Riu, A., McCollum, C. W., Knudsen, T. B., ... & Gustafsson, J. Å. Identification of estrogen target genes during zebrafish embryonic development through transcriptomic analysis. PloS one. 2013; 8(11).

Volkova, K., Caspillo, N. R., Porseryd, T., Hallgren, S., Dinnétz, P., & PorschHällström, I. Developmental exposure of zebrafish (Danio rerio) to 17αethinylestradiol affects non-reproductive behavior and fertility as adults, and increases anxiety in unexposed progeny. Hormones and behavior. 2015; 73: 30-38.

Dong, W., Teraoka, H., Yamazaki, K., Tsukiyama, S., Imani, S., Imagawa, T., Stegeman, J. J., Peterson, R. E., and Hiraga, T. 2,3,7,8-Tetrachlorodibenzo-p-dioxin toxicity in the zebrafish embryo: Local circulation failure in the dorsal midbrain is associated with increased apoptosis. Toxicol Sci. 2002; 69: 191–201.

Souder, J. P., & Gorelick, D. A. Quantification of estradiol uptake in zebrafish embryos and larvae. Toxicological Sciences. 2017; 158(2): 465-474.

Kishida , M. , McLellan , M. , Miranda, J. A . , & Callard , G . V . Estrogen and xenoestrogens upregulate the brain aromatase isoform (P450aromB) and perturb markers of early development in zebrafish (Danio rerio). Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology. 2001; 129(2): 261-268.

OECD (Organization for Economic Cooperation and Development) 236. 2006: OECD guideline for the testing of chemicals: Fish Embryo Toxicity (FET) Test. Organization for Economic Cooperation and Development, Paris, France.

MacPhail , R. C. , Brooks, J. , Hunter , D. L. , Padnos , B. , Irons, T. D. , & Padilla, S. Locomotion in larval zebrafish: influence of time of day, lighting and ethanol. Neurotoxicology. 2009; 30(1): 52-58.

Nagel, R. DarT: the embryo test with the zebrafish Danio rerio—a general model in ecotoxicology and toxicology. Altex. 2002; 19(Suppl 1): 38-48.

Yang, Y., Ma, H., Zhou, J., Liu, J., & Liu, W. Joint toxicity of permethrin and cypermethrin at sublethal concentrations to the embryo-larval zebrafish. Chemosphere. 2014; 96: 146-154.

Guiney, P. D., Walker, M. K., Spitsbergen, J. M., and Peterson, R. E. Hemodynamic dysfunction and cytochrome P4501A mRNA expression induced by 2,3,7,8 tetrachlorodibenzo-p-dioxin during embryonic stages of lake trout development. Toxicol. Appl. Pharmacol. 2000;168: 1–14.

Hagedorn , M. M., Kleinhans, F. W., Freitas, R., Liu, J., Hsu, E., Wildt, D. E., & Rall, W. F. Water Distribution and Permeability of Zebrafish Embryos (Brachydanio Rerio). Journal of zebrafish. 1997; 65(3): 199-217.

Hill, A. J., Bello, S. M., Prasch, A. L., Peterson, R. E., & Heideman, W. Water permeability and TCDD-induced edema in zebrafish early-life stages. Toxicological Sciences. 2004; 78(1): 78-87. Li, Z., Ptak, D., Zhang, L., Walls, E. K., Zhong, W., & Leung, Y. F. Phenylthiourea specifically reduces zebrafish eye size. PLoS One. 2012; 7(6): e40132.

Parker, B., & Connaughton , V. P. Effects of nicotine on growth and development in larval zebrafish. Zebrafish. 2007; 4(1): 59-68. https://doi: 10.1089/zeb.2006.9994

Pinazo –Durán, M. D., Pons-Vázquez, S., Gallego-Pinazo, R., Estrada, C. G., Zanón-Moreno, V., Bou, V. V., & Solana, P. S. Thyroid hormone deficiency disrupts rat eye neurodevelopment. Brain research. 2011; 1392: 16-26.

Takabayashi, S., Umeki, K., Yamamoto, E., Suzuki, T., Okayama, A., & Katoh, H. A novel hypothyroid dwarfism due to the missense mutation Arg479Cys of the thyroid peroxidase gene in the mouse. Molecular Endocrinology. 2006; 20(10): 25842590.

Bailey, J., Oliveri, A., & Levin, E. D. Zebrafish model systems for developmental neurobehavioral toxicology. Birth Defects Research Part C: Embryo Today: Reviews. 2013; 99(1): 14-23.

D’Amico, L. J., Li, C. Q., Seng, W. L., & McGrath, P. Developmental neurotoxicity assessment in zebrafish: a survey of 200 environmental toxicants. Society of Toxicology. 2008; 17(8): 845-869.

Emran, F., Rihel, J., Adolph, A. R., Wong, K. Y., Kraves, S., & Dowling, J. E. OFF ganglion cells cannot drive the optokinetic reflex in zebrafish. Proceedings of the National Academy of Sciences. 2007; 104(48): 19126-19131.

Burgess, H. A., & Granato, M. Modulation of locomotor activity in larval zebrafish during light adaptation. Journal of Experimental Biology. 2007; 210(14), 2526-2539.

Thompson, R. F., & Spencer, W. A. Habituation: a model phenomenon for the study of neuronal substrates of behavior. Psychological review. 1966; 73(1): 16.

Guo , S. Linking genes to brain, behavior and neurological diseases: what can we learn from zebrafish?. Genes, Brain and Behavior. 2004; 3(2): 63-74.

Kokel, D. , Bryan, J. , Laggner, C., White, R., Cheung, C. Y. J., Mateus, R., ... & MacRae, C. A. Rapid behavior-based identification of neuroactive small molecules in the zebrafish. Nature chemical biology. 2010; 6(3): 231.

Jin, Y., Wang, W., Sheng, G. D., Liu, W., & Fu, Z. Hepatic and extrahepatic expression of estrogen-responsive genes in male adult zebrafish (Danio rerio) as biomarkers of short-term exposure to 17β-estradiol. Environmental monitoring and assessment. 2008; 146(1-3): 105-111.

Brand, M., Heisenberg, C. P., Warga, R. M., Pelegri, F., Karlstrom, R. O., Beuchle, D., ... & Granato, M. Mutations affecting development of the midline and general body shape during zebrafish embryogenesis. Development. 1996; 123(1): 129-142.

Sárria, M. P., Soares, J., Vieira, M. N., Castro, L. F. C., Santos, M. M., & Monteiro, N. M. Rapid-behaviour responses as a reliable indicator of estrogenic chemical toxicity in zebrafish juveniles. Chemosphere. 2011; 85(10): 1543-1547

Copyright (c) 2020 Journal of Biomedical and Clinical Sciences (JBCS)

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Journal of Biomedical and Clinical Sciences (JBCS) publishes original research articles, short review, short communication, case reports, quiz and letter to editor that covering the areas of fundamental aspects of biomedical and clinical sciences, from pre-clinical towards translational research and their applications in medicine, dentistry and healthcare management. JBCS aims to provide a platform for basic science and clinical researchers to communicate and share their research findings to the advancement of technologies in biomedical and clinical sciences by providing maximum access to scholarly communication.

Flag Counter           


                                              Copyright © 2016 AMDI Publisher, Universiti Sains Malaysia.
Disclaimer : This website has been updated to the best of our knowledge to be accurate. However, Universiti Sains Malaysia shall not be liable for any loss or damage caused by the usage of any information obtained from this web site.
                                            Best viewed: Mozilla Firefox 4.0 & Google Chrome at 1024 × 768 resolution.