Liu Hai-Jun, Ma Jing-Hua, Liu Ru-Ming, Liu Zhang-Guo, Huang Hai-Jun, Zou Jian-Qiang, Liu Jian-Xun, Zhang Xian-Fu
College of Animal Science and Technology, Zhejiang Agriculture & Forestry University, Hangzhou, Zhejiang, China.
Hangzhou Safari Park, Hangzhou, Zhejiang, China.
Anim Reprod. 2022 Jul 4;19(2):e20220034. doi: 10.1590/1984-3143-AR2022-0034. eCollection 2022.
The objective of this study was to evaluate the fertilization capability of White Bengal Tiger frozen-thawed completely immotile spermatozoa after interspecific intracytoplasmic sperm injection (ICSI) with bovine oocytes. The fertilization status of presumptive zygotes was assessed 18 h after ICSI by immunofluorescence staining and confocal microscopy. The fertilization rate was 34.8% (8/23), as confirmed by the extrusion of two polar bodies, or male and female pronuclei formation. For unfertilized oocytes (65.2%, 15/23), one activated oocyte had an activated spermatozoon but most were unactivated oocytes with unactivated spermatozoa (1/15, 6.7% vs 10/15, 66.7%, respectively, < 0.05). These results showed that White Bengal Tiger frozen-thawed completely immotile spermatozoa retained the capacity to fertilize bovine oocytes after interspecific ICSI. This is the first report of in vitro produced zygotes using tiger immotile sperm with bovine oocytes by interspecific ICSI technique, which provides an efficient and feasible method for preservation and utilization of endangered feline animals.
本研究的目的是评估白孟加拉虎冷冻解冻后完全不动的精子在与牛卵母细胞进行种间胞浆内单精子注射（ICSI）后的受精能力。在ICSI后18小时，通过免疫荧光染色和共聚焦显微镜评估推定受精卵的受精状态。通过两个极体的排出或雌雄原核的形成证实，受精率为34.8%（8/23）。对于未受精的卵母细胞（65.2%，15/23），一个激活的卵母细胞有一个激活的精子，但大多数是未激活的卵母细胞和未激活的精子（分别为1/15，6.7%对10/15，66.7%，<0.05）。这些结果表明，白孟加拉虎冷冻解冻后完全不动的精子在种间ICSI后仍保留使牛卵母细胞受精的能力。这是首次报道通过种间ICSI技术使用老虎不动精子与牛卵母细胞体外产生合子，为濒危猫科动物的保存和利用提供了一种有效且可行的方法。
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