【作者】 向红；

【导师】 纳冬荃； 刘苹；

【作者基本信息】 昆明医学院 ， 劳动卫生与环境卫生学， 2003， 硕士

【摘要】 目的 探索家兔超数排卵的影响因素和受精卵显微穿刺操作的适宜条件。方法 采用FSH+HCG激素组合的处理方法，比较性周期不同阶段、品种和季节变化对家兔超数排卵的影响；将受精卵暴露于体外(23℃、1800lx)不同时间段后观察其体外培养的发育情况；用含0.02％胰酶的水解乳蛋白液洗胚，分析不同洗胚时间受精卵的卵裂率；在显微穿刺操作过程中，比较不同粗细穿刺针，不同的穿刺时间对受精卵体外发育的影响。结果家兔发情期或发情后期超数排卵数明显比非发情期高(P＜0.05)，日本大耳兔超数排卵数(38.8±10.6)比本地家兔的超数排卵数(30.0±5.7)高，昆明地区的季节变化对家兔超数排卵未见有明显的影响；受精卵暴露于体外30分钟内，体外培养8小时卵裂率、48小时桑葚胚率与对照组一致(P＞0.05)；用0.02％胰酶水解乳蛋白液洗胚3分钟为最佳时间；用直径小于1μm注射针穿刺兔受精卵原核后体外培养8小时卵裂率(89.2％)与对照组(92.0％)一致(P＞0.05)；受精卵穿刺时间在10分钟内，48小时培养8—细胞期以上卵裂率为80.2％，与对照组(85.8％)相近。结论 日本大耳兔在发情期或发情后期超排卵数较多，兔受精卵在体外暴露30分钟内、0.02％胰酶的水解乳蛋白液洗3分钟、用小于1μm穿刺针以及10分钟内完成显微穿刺操作对其体外发育能力无明显影响。更多还原

【Abstract】 Objective: To study the influential factors of superovulation and the appropriate condition of microinjection manipulation for zygotes. Methods: Comparing the effect of sex cycle, breed and seasons on superovulation of female rabbits after they were treated with FSH and HCG. Evaluating the development of embryos after they were exposed to room temperature (23 degrees C ) and visible light (18001x) for various lengths of time. Analyzing the cleavage rate of zygotes after they were washed in 0.02% insulinase lactalbumin hydrolysate solution for different lengths of time. Exploring the influence of the size of micropipette and the different length of time of injection on the survival rate of embryos in microinjection procedure. Result: The average number of superovulation in rabbits during estrum was higher than that during no estrum(P<0.05). The average number of embryos was 38±10.6 in Japan white rabbits, 30.0±5.7 in native white rabbits. The influence of season changes on superovulation was not obvious (P>0.05) in Kunming. The 8 hour 2-cell cleavage rate and 48 hour morula rate in vitro culture were comparable to that of control group when the exposed time of eggs was less than 30min. The appropriate time for washing embryos in 0.02% insulinase lactalbumin hydrolysate solution was 3min. There was no significant difference between trial group and control group in terms of cleavage rate when the size of micropipette is less than 1μm. When the length of ume of microinjection is 10min, 8-cell-morula cleavage rate for 48hr in vitro culture was 80.2% , as well as the control group. Conclusion: Japanwhite rabbits treated with FSH and HCG during estrum can harvest more embryos. The development of embryos in vitro was not obviously affected when they were exposed outside for 30min, washed in 0.02% insulinase lactalbumin hydrolysate solution for 3min, pricked with the size of pipette within 1μm and took 10min in microinjection procedure.更多还原