Working Paper Article Version 1 This version is not peer-reviewed

Optimization of Sperm Storage and Fertilization in Zebrafish [Danio rerio (Hamilton)]

Version 1 : Received: 26 November 2020 / Approved: 27 November 2020 / Online: 27 November 2020 (10:11:39 CET)

How to cite: Cheng, Y.; Franěk, R.; Rodina, M.; Xin, M.; Cosson, J.; Linhart, O. Optimization of Sperm Storage and Fertilization in Zebrafish [Danio rerio (Hamilton)]. Preprints 2020, 2020110682 Cheng, Y.; Franěk, R.; Rodina, M.; Xin, M.; Cosson, J.; Linhart, O. Optimization of Sperm Storage and Fertilization in Zebrafish [Danio rerio (Hamilton)]. Preprints 2020, 2020110682

Abstract

The zebrafish Danio rerio is suitable to study gametes as a model organism. There were > 70% of zebrafish spermatozoa activated, because they were contaminated with urine or excrement. The movement of spermatozoon in water was propagated along the flagellum at 16 s after sperm activation, then damped from the end of the flagellum for 35 s and fully disappear at 61 s after activation. For artificial fertilization, milt must be added to an extender, which stops the movement of sperm and keeps the sperm motionless until fertilization. E400 was shown to be the most suitable extender as it allows to store sperm for fertilization for 6 to 12 h at 0-2oC. Sperm motility decreased only to 36% at 12 h post stripping (HPS) for E400 extender and to 19% for Kurokura extender. To achieve an optimal level of fertilization and hatching, a test tube with a well-defined amount of 6,000,000 spermatozoa in E400 extender per 100 eggs and 100 µl of activation solution has proved to be more successful than using a Petri dish. The highest fertilization and hatching rates reached 80% and 40-60%, respectively, with milt stored for 1.5 h in E400 extender at 0-2oC.

Subject Areas

zebrafish; Danio rerio; sperm motility; fertilization; short-term storage; extender

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