TY  - JOUR
TI  - Room-Temperature Storage of Zebrafish and Medaka Sperm Using Lactic Acid-Stabilized L-15 Medium
AU  - Imai, Yukiko 
AU  - Asano, Touma 
AU  - Nishimura, Toshiya 
AU  - Takemoto, Kazumasa 
AU  - Kawamura, Akinori 
AU  - Sakai, Noriyoshi  
VL  - 15
IS  - 23
PY  - 2025
DA  - 2025/12/05
SP  - e5534
C1  - Bio-protocol 2025;15:e5534
DO  - 10.21769/BioProtoc.5534
UR  - https://doi.org/10.21769/BioProtoc.5534
AB  - Zebrafish offer numerous advantages as a vertebrate model because of their rapid development, high fecundity, transparent embryos, and ease of genetic manipulation. A wide variety of transgenic and mutant fish lines have been generated, and efficiently sharing these resources is crucial for advancing research. Zebrafish lines have typically been exchanged as early embryos, adult fish, or cryopreserved sperm, making transportation costly and logistically challenging. Here, we provide a protocol for preserving functional zebrafish sperm for more than 7 days at room temperature and subsequent in vitro fertilization using the preserved sperm. In this protocol, sperm collected either from the cloaca of an anesthetized male or from dissected testes is stored in L-15-based storage medium. Importantly, the storage medium, originally developed for zebrafish, is also applicable to medaka, another widely used vertebrate model. This sperm storage method allows researchers to ship sperm using low-cost methods and to investigate key factors for motility and fertilizing ability in those sperm.
KW  - Zebrafish
KW  - Medaka
KW  - Sperm
KW  - In vitro fertilization
KW  - Bioresource
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False