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Room-Temperature Storage of Zebrafish and Medaka Sperm Using Lactic Acid-Stabilized L-15 Medium.

Yukiko Imai1, Touma Asano1, Toshiya Nishimura2

  • 1Division of Life Science, Graduate School of Science and Engineering, Saitama University, Saitama, Japan.

Bio-Protocol
|December 12, 2025
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Summary

Researchers can now preserve zebrafish and medaka sperm at room temperature for over a week. This breakthrough simplifies fish line sharing through cost-effective shipping and successful in vitro fertilization.

Keywords:
BioresourceIn vitro fertilizationMedakaSpermZebrafish

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Area of Science:

  • Aquatic biology
  • Developmental biology
  • Genetics

Background:

  • Zebrafish are valuable vertebrate models due to their rapid development, high fecundity, and genetic manipulability.
  • Efficient sharing of transgenic and mutant zebrafish lines is essential for research advancement.
  • Current methods for exchanging fish lines (embryos, adults, cryopreserved sperm) are costly and logistically difficult.

Purpose of the Study:

  • To develop a protocol for preserving functional zebrafish sperm at room temperature for extended periods.
  • To enable successful in vitro fertilization using preserved sperm.
  • To facilitate low-cost, international exchange of zebrafish and medaka lines.

Main Methods:

  • Sperm collection from anesthetized male zebrafish (cloaca or dissected testes).
  • Storage of sperm in an L-15-based medium at room temperature.
  • In vitro fertilization trials using preserved sperm samples.

Main Results:

  • Functional zebrafish sperm preserved for over 7 days at room temperature.
  • The storage medium is also effective for medaka sperm preservation.
  • High fertilization rates achieved using room-temperature preserved sperm.

Conclusions:

  • A novel protocol enables room-temperature sperm preservation for zebrafish and medaka, lasting over a week.
  • This method significantly reduces the cost and logistical challenges of exchanging fish lines.
  • The protocol supports global research collaboration by simplifying the sharing of valuable genetic resources.