【摘要】 本实验用化学药物对家兔胚胎细胞周期进行调控 ,以建立细胞周期同期化的方法。实验结果表明 :(1)用 1 0 0 μg/ml噻氨酯哒唑 (nocodazole)处理家兔 16 细胞胚胎 12h ,可使胚胎细胞完全同期化于M期 ;(2 )细胞周期同期化于M期的家兔胚胎细胞用 0 2 0 μg/ml阿菲迪霉素 (aphidi colin)处理 8h ,可完全抑制DNA的合成 ,使胚胎细胞同期化于G1期 ;(3)细胞周期同期化于M期的家兔胚胎细胞在体外培养 8h后 ,然后用 1 0 0 μg/ml环己酰胺 (cycloheximide)处理 8h ,可使胚胎细胞同期化于G2 期。本研究的结果还表明细胞周期同期化处理不影响胚胎的体外发育 ,但胚胎移植后的产仔率明显低于对照组 (P <0 0 5 )。更多还原

【Abstract】 The objective of this experiment was to control the rabbit embryonic cell cycle with chemicals to give programs for the cycle synchronization.The DNA contents of blastomeres were analyzed by scanning microspectrophotometric assay following Feulgen staining reaction.The reliable methods for synchronizing cell cycle at M,G 1 and G 2 phase were given in the followings.When the early 16 cell stage rabbit embryos were cultured in M199 supplemented with 1 00 μg/ml nocodazole for 12 h,the blastomeres were all synchronized at M phase.When the embryos which cells arrested at M phase were cultured in M199 containing 0 20 μg/ml alphidicolin for 8h,the embryos were divided into 32 cell,but they did not synthesized DNA and were arrested at G 1 phase.The embryonic cells were arrested at G 2 phase after the embryos were first cultured in 1 0 μg/ml nocodazole for 12h,then in M199 for 8h and finally in 1 00 μg/ml cycloheximide for 8h.The developmental abilities of embryos synchronized at special cell cycle stage were not be effected when they were cultured in M199 in vitro, but rates of living pups were significantly lower than the control after the synchronized embryos were transferred into recipients(P<0 05).更多还原