不同条件延缓未受精卵激活对鱼类细胞电融合的影响

泥鳅未受精卵先在合成卵巢液、郝氏液中分别浸泡15、20、30min,然后再受精孵化,统计发育情况,结果表明,合成卵巢液可有效延缓未受精卵遇水后被激活的时间,郝氏液对延缓未受精卵被激活也有一定作用,但效果比合成卵巢液差。用合成卵巢液配制不同浓度胰酶处理受体卵进行细胞电融合实验,胰酶质量浓度为5 mg/mL时,融合卵发育至囊胚期的比例比对照组(郝氏液)高2%,统计分析表明该差异是显著的(P<0.05);但发育至原肠期及孵出期的比例则与对照组无显著差异。胰酶质量浓度为2和1 mg/mL时,融合卵发育到各阶段的比例均低于对照组,统计分析表明两者的差异是显著的(P<0.05)。研究表明,溶液种类、胰酶浓度等对电融合卵的发育有明显的影响,但延缓受体卵的激活时间并不能提高电融合卵的出苗率。用郝氏液配制的2 mg/mL的胰酶溶液处理受体卵,可获得较高比例的融合鱼。

Effect on Fish Cell Electrofusion by Delayed Activation Time for Unfertilized Oocytes under Different Conditions

Unfertilized oocytes of loach Praramisgurnus dabryanus were socked in synthetic ovarian fluid(SOF) and Holfreter's solution for 15,20 or 30 minutes respectively before fertilization.The results showed that SOF effective delayed activation time of oocytes after being in water;and Holfreter's solution also had some effects,but not so efficient as SOF.The oocytes,which were digested by different concentrations of Trypsin,used as recipient.Fusion experiments indicated that Trypsin whose mass concentration is 5 mg/mL in SOF,in comparison with the control group(Holfreter's solution),noticeably raised the development rate of fused eggs to blastula stage.The statistical analysis showed the difference is obvious(P<0.05).The ratios of fused oocytes developed to gastrula and hatching stages,however,were not so different with the control group.In addition,when the mass concentration of Trypsin were 2 and 1 mg/mL,the ratios of fused oocytes developed to various stages were all lower than those of the control group,the statistical analysis indicated the differences were significant(P<0.05).This paper demonstrated that the types of solutions and concentrations of Trypsin have obvious effects on the development of electrofusion oocytes,but the delayed activation time of recipient oocytes would not raise the hatching rate.The recipient oocytes treated with 2 mg/mL Trypsin administrated by Holfreter's solution might achieve higher rate of fused fish.