Comparison between PMSG- and FSH-induced superovulation for the generation of transgenic rats.
Superovulation protocols using single injections of pregnant mare's serum gonadotropin (PMSG) or minipumps with follicle-stimulating hormone (FSH) were compared in immature Sprague-Dawley (SD) rats. We used the following criteria: total number of ova, rate of fertilization, in vitro embryo development, sensitivity of zygotes to the microinjection of foreign DNA into the pronucleus, and their in-vivo development after transplantation into the oviduct of a recipient. Female SD rats were stimulated with 15 IU PMSG or 10 mg FSH followed by the injection of human chorionic gonadotropin (hCG) at doses of 20 and 30 IU per female. After hCG administration, they were mated with males of the same strain and sacrificed on day 1 of pregnancy. The percentage of mated animals and the fertilization rate was similar in all groups. In rats given PMSG, the number of ovulated zygotes was hCG dose-dependent. In contrast, the dose of hCG did not influence the efficiency of superovulation in rats given FSH, which was equal to PMSG-treated rats at the optimal dose of hCG. The rates of in vitro blastocyst development (31.4 and 23.3%) and the resistance to microinjection into the pronucleus did also not differ significantly between zygotes of both studied groups. The proportion of offspring developing from microinjected zygotes after oviduct transfer (26.2 and 26.8%, respectively) and the rate of transgene integration per newborns (7.3 and 4.9%, respectively) was similar in both experimental groups. The results of this study demonstrate that superovulation of immature SD rats by PMSG is equally effective as FSH treatment and, thus, preferable for transgenic rat technology due to the lower costs and easier handling.
Max-Delbrück Center for Molecular Medicine, Berlin-Buch, and Deutsches Institut für Bluthochdruckforschung, Heidelberg, Germany.
Elena
E
Popova
Alexander
A
Krivokharchenko
Detlev
D
Ganten
Michael
M
Bader
D000818:Animals; D030801:Animals, Genetically Modified; D005260:Female; D005306:Fertilization / Q000187:drug effects; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D018014:Gene Transfer Techniques; D006064:Gonadotropins, Equine / Q000494:pharmacology*; D008845:Microinjections; D010063:Ovum / Q000187:drug effects; D051381:Rats; D012726:Sexual Behavior, Animal / Q000187:drug effects; D013480:Superovulation / Q000187:drug effects*; D015053:Zygote / Q000187:drug effects / Q000254:growth & development
D006064:Gonadotropins, Equine; D005640:Follicle Stimulating Hormone