Hewitson L C, Simerly C R, Tengowski M W, Sutovsky P, Navara C S, Haavisto A J, Schatten G
Department of Zoology, University of Wisconsin, Madison 53706, USA.
Biol Reprod. 1996 Aug;55(2):271-80. doi: 10.1095/biolreprod55.2.271.
Intracytoplasmic sperm injection (ICSI) was performed on rhesus monkey oocytes, and the resultant microtubule and DNA configurations were imaged by laser-scanning confocal microscopy. In addition, polyspermic oocytes fertilized by ICSI were examined by transmission electron microscopy (TEM). Successful rhesus fertilization by ICSI revealed microtubule and DNA configurations similar to those observed during in vitro fertilization of human and rhesus monkey oocytes, including sperm aster formation, pronuclei decondensation, spindle formation, and cell division. Several abnormalities, however, were also observed: 1) inability to complete meiosis; 2) inability to undergo male or female pronucleus formation; 3) separation of the sperm tail from the sperm nucleus; 4) premature chromosome condensation with the formation of a paternal meiotic spindle; and 5) formation of multiple female pronuclei (karyomeres) during chromosome decondensation. TEM analysis revealed that sperm can undergo decondensation in the presence of an intact acrosome at least 18 h after sperm injection. These results demonstrate the utility of rhesus ICSI in pre-clinical applications as well as with endangered species. However, the different types of fertilization failures observed here indicate that although ICSI may be a readily accepted means of fertilization of human oocytes in many clinics, we should further characterize the cellular and genetic abnormalities associated with ICSI in both human and nonhuman primates.
对恒河猴卵母细胞进行了胞浆内单精子注射（ICSI），并通过激光扫描共聚焦显微镜对由此产生的微管和DNA构型进行成像。此外，通过透射电子显微镜（TEM）检查了经ICSI受精的多精卵母细胞。ICSI成功使恒河猴受精，显示出与人类和恒河猴卵母细胞体外受精过程中观察到的微管和DNA构型相似，包括精子星体形成、原核解聚、纺锤体形成和细胞分裂。然而，也观察到了一些异常情况：1）无法完成减数分裂；2）无法形成雄原核或雌原核；3）精子尾部与精子核分离；4）染色体过早凝聚并形成父本减数分裂纺锤体；5）在染色体解聚过程中形成多个雌原核（核粒）。TEM分析表明，精子在注射后至少18小时，在顶体完整的情况下可以发生解聚。这些结果证明了恒河猴ICSI在临床前应用以及濒危物种方面的实用性。然而，此处观察到的不同类型的受精失败表明，尽管ICSI在许多诊所可能是人类卵母细胞受精的一种容易被接受的方式，但我们应该进一步明确人类和非人类灵长类动物中与ICSI相关的细胞和遗传异常情况。
