Delayed in vitro fertilization of zebrafish eggs in Hank's saline containing bovine serum albumin.
In zebrafish it is possible to create viable diploid fish whose genomic DNA is derived only from the female parent (parthenogenesis) or, as was more recently shown, only from the male (androgenesis). Androgenesis requires holding zebrafish eggs in an inactivated state in vitro for an hour or more. Previously this was achieved by placing the zebrafish eggs in ovary fluid obtained from rainbow trout (Onchorhynchus mykiss) or coho salmon (Onchorhynchus kisutch). Here we report that adding bovine serum albumin (BSA) to Hank's buffered saline prevents zebrafish egg activation in vitro. Of the zebrafish eggs placed in Hank's saline plus 0.5% BSA, 85% +/- 8.7% were fertilizable after incubation for one hour at room temperature (23 degrees C). Longer incubations are possible but with lower efficiency of fertilization. This technique not only could facilitate androgenesis, but also might be useful when making transgenics by microinjection, when performing antibody or RNA injections before fertilization, or for studying the mechanisms of egg activation in zebrafish.
Sakai
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N
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139 USA.
Burgess
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S
Hopkins
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N
D000818:Animals; D005260:Female; D005306:Fertilization / Q000502:physiology*; D008297:Male; D010063:Ovum / Q000187:drug effects*; D010312:Parthenogenesis; D012710:Serum Albumin, Bovine / Q000494:pharmacology*; D012965:Sodium Chloride; D015027:Zebrafish / Q000502:physiology*
D012710:Serum Albumin, Bovine; D012965:Sodium Chloride