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Azole pharmaceuticals induce germinal vesicle breakdown (GVBD) in preovulatory oocytes of zebrafish (Danio rerio): an in vitro study

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Abstract

Azoles, the antifungal pharmaceuticals are emerging as a new class of water contaminants with a potential to influence the endocrine physiology of surrounding aquatic fauna. In this study, we made an attempt to assess the relative efficacy of widely used azoles belonging to two subclasses, i.e., (i) triazoles (letrozole, fluconazole, itraconazole) and (ii) imidazoles (ketaconazole, ornidazole, clotrimazole), on the onset of germinal vesicle breakdown (GVBD) (an initial step in the final maturation of oocytes) in fully grown preovulatory oocytes of zebrafish (Danio rerio) using an in vitro model. Oocytes (> 650 μm) isolated manually from gravid ovaries were exposed to (i) 0.01 and/or 0.1, 1.0, 5.0, 10, 15, and 20 ng/ml and (ii) 1.0, 2.0, 3.0, 4.0, and 5.0 μg/ml of drugs. Zebrafish Ringer’s solution (vehicle) and 0.01% ethyl alcohol (solvent) were used as negative controls. 17α, 20 β-Dihydroxy-4-pregnen-3-one (17α-DHP) and diethylstibestrol (DES), potent inducers of GVBD in fish, were used as positive controls. GVBD was scored hourly from 0–6 h. In negative controls, there were no indications of GVBD even at the 6th hour, while in 17α-DHP- and DES-exposed oocytes, GVBD was initiated from the 1st hour, reaching 80% and 76% respectively at the 6th hour. Among azoles, letrozole induced GVBD in 73–85%, fluconazole (30–33%), itraconazole (23–33%), ketaconazole (46–53%), ornidazole (36–40%), and clotrimazole (30–33%) of oocytes. These results suggest that azole pharmaceuticals induce GVBD in fish oocytes that may be attributed to their variable degree of cytochrome P450 enzyme inhibitor activity.

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All data generated or analyzed during this study are included in this published article.

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Acknowledgments

We thank Ms. Vani Joshi, Ms. Shubha Markande, and Ms. Madhuri Kanetkar for assisting in conducting the experiments and analysis of the data.

Funding

The work is supported by UGC-SAP-DSA-I Program of Dept. of Zoology, KUD. BBG gratefully acknowledges UGC, New Delhi, for grant of BSR-RFSMS fellowship.

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  1. Department of Zoology, Karnatak University, Dharwad, 580003, India

    Pancharatna A. Katti & Basavaraj B. Goundadkar

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  1. Pancharatna A. Katti
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  2. Basavaraj B. Goundadkar
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KP conceived and designed the experiments and wrote the paper; BG performed the experiments, analyzed the data, and prepared the graphs and manuscript.

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Correspondence to Pancharatna A. Katti.

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Permission to work on zebrafish was obtained from the Animal Ethical Committee (Committee for the Purpose of Control and Supervision of Experiments on Animals—CPCSEA), Government of India, under Institutional Registration #639/GO/02/a/CPCSEA.

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Katti, P.A., Goundadkar, B.B. Azole pharmaceuticals induce germinal vesicle breakdown (GVBD) in preovulatory oocytes of zebrafish (Danio rerio): an in vitro study. Environ Sci Pollut Res 28, 3694–3702 (2021). https://doi.org/10.1007/s11356-020-10719-x

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