本研究的目的是调查精子的自发运动和优化精子收集,短期精子储存,斑马鱼Daniorerio受精.精子在水中的运动在精子激活后的16s沿鞭毛传播,然后从鞭毛末端衰减35s,并在激活后的61s完全消失。对于人工施肥,必须将milt添加到固定溶液中,停止精子的运动,并保持精子不动,直到受精。E400和Kurokura作为等渗溶液被证明是储存精子用于受精6小时的合适补充剂。E400在0-2°C下储存精子12小时。E400扩展器的精子活动力在剥离后12小时仅下降至36%,而Kurokura扩展器的精子活动力下降至19%。为了达到最佳的受精水平和游动幼虫率,事实证明,每100个卵中含有6,000,000个精子的E400补充剂和100µL活化溶液的试管比使用培养皿更成功。最高的受精率和游动幼虫率分别达到80%和40-60%,分别,在0-2°C下,在E400扩展器中储存1.5h。
The aim of the present study was to investigate the spontaneous motility of spermatozoa and to optimize sperm collection, short-term sperm storage, and fertilization in zebrafish Danio rerio. The movement of spermatozoon in water was propagated along the flagellum at 16 s after sperm activation then damped from the end of the flagellum for 35 s and fully disappeared at 61 s after activation. For artificial fertilization, milt must be added to an immobilizing solution, which stops the movement of sperm and keeps the sperm motionless until fertilization. E400 and Kurokura as isotonic solutions were shown to be suitable extenders to store sperm for fertilization for 6 h. E400 stored sperm for 12 h at 0-2 °C. Sperm motility decreased only to 36% at 12 h post stripping for the E400
extender and to 19% for the Kurokura
extender. To achieve an optimal level of fertilization and swim-up larvae rates, a test tube with a well-defined amount of 6,000,000 spermatozoa in E400
extender per 100 eggs and 100 µL of activation solution has proven to be more successful than using a Petri dish. The highest fertilization and swim-up larvae rates reached 80% and 40-60%, respectively, with milt stored for 1.5 h in the E400
extender at 0-2 °C.