微注射MT-hGH基因入雄原核后兔受精卵体外发育的研究

Study on Development of Fertilized Rabbit Ova in vitro Following Microinjeetion of MT-hGH Gene in to Male Pronucleus

将MT-hGH基因用微注射的方法,注入兔原核卵的雄原核中,微注射的受精卵和不经微注射对照的受精卵,经体外培养9h,21h,33h,53h,77h和125h,对其活力,发育速率和发育阶段进行观察。其结果是对照组和注射组的存活率随着体外培养时间的延长而下降,对照组受精卵体外培养9h存活率为99.2％,到125h下降至68.64％;注射组受精卵体外培养9h存活率为98.2％到125h下降至62.1％,体外培养9h和体外培养125h,胚胎存活率差异极显著(P<0.01)。在相同的时间内对照组胚胎存活率略高于微注射组,两者差异不显著。体外培养胚胎的发育速率对照组和微注射组均低于体内发育,而体外培养对照组胚胎发育速率与微注射组比较无显著差异,前者略高于后者。将注射MT-hGH基因体外培养发育正常的胚胎,移入8只同步假孕受体输卵管中,3只受体妇娠期满产仔5只,目前4只仔兔生长良好,其月令体重相当于对照仔兔1.3倍。注入外源基因整合情况有待检测后另文发表。

Fertilized rabbit ova were microinjccted MT-hGH gene into male pronucleus.Microinjccted ova and not injected control ova were cultured in vitro.The survival potency,developmental speed and developmental stage of these ova were observed at different times.After 9 hours,21hr,33hr,53hr,77hr and 125hr culture in vitro,survival rate of microinjccted and control embryos were observed. After 9 hours culture in vitro,survived rate of control embryos were 99.2%, after 125hr culture in vitro was 68.64%.Survived rate of microinjected embryos after 9hr cultrue in vitro was 98.2%,125hr was 62.1%. Survived rate of embryos eultrue in vitro remarkable decreased at differnet times.Survived rate of microinjected embryos lower than control,but did not remarkable differ at the same times from the culture in vitro.Developmental speed of embryos cultured in vitro was lower than vivo.There is not difference between developmental speed of embryos in vitro controled and mieroinjected.After 4-5hr culture in vitro microinjected embryos of normal development were transffered surgically to the oviducts of 8 synchronized pseudopregnant recipients.3 recipients bring 5 offsprings.Up to now 4 offsprings survived and theirs boby weight were 1.3 times more than control.