Ohsumi K, Katagiri C, Yanagimachi R
J Exp Zool. 1986 Mar;237(3):319-25. doi: 10.1002/jez.1402370304.
Human spermatozoa were demembranated with Triton X-100 (TX) and injected into the mature eggs of Xenopus laevis. The nuclei of these spermatozoa decondensed and developed into pronuclei. Chromosomes did not appear in the eggs until the end of a 5-hr incubation period. When the demembranated human spermatozoa were further treated with dithiothreitol (DTT) before they were injected into the eggs, the sperm nuclear decondensation and pronuclear development took place considerably faster than in spermatozoa treated with the detergent alone. By the end of the 5-hr incubation period, decondensed chromatin threads or chromosome-like structures appeared, but none of the eggs cleaved. When human spermatozoa were injected into full-grown ovarian oocytes with intact germinal vesicle (GV) or oocytes which had matured without GV, the nuclei of a proportion of TX-treated and all TX-DTT-treated sperm decondensed but showed no sign of developing into pronuclei. Sperm nuclei injected into maturing oocytes formed condensed chromatin fragments as long as the oocytes were not activated, but they transformed into pronuclei when the oocytes were stimulated with electric shock. These results indicate that the cytoplasmic factors responsible for the decondensation of human sperm nuclei are present in egg cytoplasm independent of GV-materials. We also suggest that the factors controlling development of decondensed sperm nuclei into pronuclei are dependent on GV materials.
用曲拉通X - 100(TX)去除人精子的细胞膜,并将其注入非洲爪蟾的成熟卵母细胞中。这些精子的细胞核去浓缩并发育成原核。直到5小时孵育期结束,卵母细胞中才出现染色体。当去除细胞膜的人精子在注入卵母细胞之前先用二硫苏糖醇(DTT)进一步处理时,精子核去浓缩和原核发育的速度比仅用去污剂处理的精子要快得多。在5小时孵育期结束时,出现了去浓缩的染色质丝或染色体样结构,但没有一个卵母细胞分裂。当将人精子注入具有完整生发泡(GV)的完全成熟的卵巢卵母细胞或未经过GV而成熟的卵母细胞时,一部分经TX处理和所有经TX - DTT处理的精子的细胞核去浓缩,但没有发育成原核的迹象。只要卵母细胞未被激活,注入成熟卵母细胞的精子核就会形成浓缩的染色质片段,但当卵母细胞受到电击刺激时,它们会转化为原核。这些结果表明,负责使人精子核去浓缩的细胞质因子存在于卵细胞质中,与GV物质无关。我们还认为,控制去浓缩精子核发育成原核的因子依赖于GV物质。