Initial Study Of Capacitation On Blue Fox Sperm In Vitro

The capacitation of sperm in vitro is the basic process of the In vitro fertilization (â…£) and Embryo transplant (ET). To work over the capacitation of sperm in vitro is with great significance of the reproduction study. In the experiment, the swim-up was used for select sperms. The experiment was divided for there parts. The first, the fresh ejaculated semen of the blue fox was incubated with the substrate of mTyrod's solution (group T), BO solution(B) and the high ionic strength medium (H) at 38.5℃in a 5% CO₂ atmosphere. The second, the sperm was incubated in BO solution with the different level of temperature of 37.5℃(group B1),38.5^C(B2),39.5℃(B3) in a 5% CO₂ atmosphere. The last, the 0,10,20,50μg/ml of heparin was add to the solution of mTyrod's respectively at 38.5℃in a 5% CO₂ atmosphere(group T1-T4). The time for the incubation of the capacitation were last for 6 hours. Sperm quality in blue fox during the capacitation in vitro was evaluated by examing viability with eosin-nigrosine staining, examing the acrosome reaction rate by using Coomassie blue staining, and observing the activity of the sperms at the 0,1,2,4,6 hour of the incubation for the capacitation for 4 repeats.The result shows that the higher acrosome reaction rate was the lower activity the sperms become. For the first part, in the incubation of the capacitation for 6 hours, the sequence of the acrosome reaction rate for group T,B,H were H＞B＞T (P＜0.05); the viability rate of group T,B,H were T＞H＞B. These results indicated that the BO solution(B) is better than other solutions on protecting sperms. For the second part, the results showed that the acrosome reaction rate of group B2 was 72.49±5.26%, which was higher than the other groups notably(P＜0.05); the sequence of the viability rate for group B1,B2,B3 were B1＞B3＞B2; the change of the activity of the sperms were B2＞B1＞B3. That means when the acrosome reaction take place the activity of the sperms will become lower. The 38.5℃is suit for the capacitation in vitro of the blue fox. For the last part, the 0,10,20,50μg/mi of the heparin was named group T1,T2,T3,T4 for experiment. The sequence of the acrosome reaction rate for group T1,T2,T3,T4 were T3＞T2＞TI＞T4; the acrosome reaction rate of group T3 was 68.57±8.03% which was higher than the other groups notably(P＜0.05); the sequence of the viability rate for these groups were also T3＞T2＞T1＞T4(P＞0.05); the change of the activity of the sperms were T2,T3＞T1＞T4(P＞0.05). The results showed that the suitable consistence of the heparin was 20μg/ml.On the foundation of these experiment, the thesis conclude that: the BO solution(B) is better than the mTyrod's solution and the high ionic strength medium; the suitable time of incubation for the capacitation was t=6 hour; the high ionic strength medium was good for the capacitation, but high ionic strength was bad for the preservation of sperm; The 38.5℃is suit for the capacitation in vitro of the blue fox; and the suitable consistence of the heparin was 20μg/ml.