Publication Date:
2022-05-26
Description:
© The Author(s), 2017. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Theriogenology 92 (2017): 149–155, doi:10.1016/j.theriogenology.2017.01.007.
Description:
Cryogenic storage of sperm from genetically altered Xenopus improves cost effectiveness and animal welfare associated with their use in research; currently it is routine for X. tropicalis but not reliable for X. laevis. Here we compare directly the three published protocols for Xenopus sperm freeze-thaw and determine whether sperm storage temperature, method of testes maceration and delays in the freezing protocols affect successful fertilisation and embryo development in X. laevis. We conclude that the protocol is robust and that the variability observed in fertilisation rates is due to differences between individuals. We show that the embryos made from the frozen-thawed sperm are normal and that the adults they develop into are reproductively indistinguishable from others in the colony. This opens the way for using cryopreserved sperm to distribute dominant genetically altered (GA) lines, potentially saving travel-induced stress to the male frogs, reducing their numbers used and making Xenopus experiments more cost effective.
Description:
The EXRC is supported by the Wellcome Trust (101480/Z), BBSRC (BB/K019988/1) and NC3Rs (NC/P001009/1). The NXR is supported by a grant from the NIH (P40 OD010997).
Keywords:
Xenopus
;
Sperm
;
Cryopreservation
;
Stock centres
;
Genetically altered lines
;
3Rs
Repository Name:
Woods Hole Open Access Server
Type:
Article
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