Cytoskeleton and Chromatin Reorganization in Horse Oocytes Following Intracytoplasmic Sperm Injection: Patterns Associated with Normal and Defective Fertilization
Cytoskeleton and Chromatin Reorganization in Horse Oocytes Following Intracytoplasmic Sperm Injection: Patterns Associated with Normal and Defective Fertilization
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胞浆内精子注射后马卵母细胞的细胞骨架和染色质重组：与正常和缺陷受精相关的模式
DOI：
10.1095/biolreprod.102.012823
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发表时间：
2003
期刊：
影响因子：
2.8
通讯作者：
M. Bevers
中科院分区：
文献类型：
--
作者：
J. Tremoleda;T. van Haeften;T. Stout;B. Colenbrander;M. Bevers
摘要卵母细胞胞浆内单精子注射(ICSI)是马卵母细胞体外受精的首选方法。然而，由于尚不清楚的原因，胚胎发育率很低。本研究的目的是检测ICSI受精或孤雌激活的马卵母细胞的细胞骨架和染色质重组。更多的卵母细胞被注射了标记有线粒体特有的生命染料的精子，以帮助识别精子对合子结构的贡献，特别是中心体。精子注射后每隔一段时间固定卵母细胞，用激光共聚焦扫描显微镜观察。在未受精的卵母细胞中，微管仅存在于第二次减数分裂中期停滞的纺锤体和第一极体。精子注射后，在精子头部附近形成了一组微管，随后变大，以至于在原核迁移和对位时，它充满了整个细胞质。在联配过程中，微管基质重组形成有丝分裂纺锤体，双亲的染色质在该纺锤体上排列。最后，在核和细胞分裂完成后，微管分散到间期的子细胞中。假注射诱导了76%的卵母细胞的孤雌激活，其特征是形成了多个细胞质微管灶，这些微管灶后来发展成围绕雌性原核的致密微管网络。在正常受精过程中，单性配子本身可以产生微管二倍体，而二倍体在正常受精过程中总是在精子头部的底部形成，这表明两种配子都有助于马合子中心体的形成。最后，25%的精子注射的卵母细胞未能完成受精，主要是由于没有激活卵母细胞(65%)，这往往伴随着精子解凝失败。综上所述，本研究表明，马受精卵亲本基因组的结合伴随着一系列整合的细胞骨架介导的事件，这些事件的失败会导致发育停滞。
Abstract Intracytoplasmic sperm injection (ICSI) is the method of choice for fertilizing horse oocytes in vitro. Nevertheless, for reasons that are not yet clear, embryo development rates are low. The aims of this study were to examine cytoskeletal and chromatin reorganization in horse oocytes fertilized by ICSI or activated parthenogenetically. Additional oocytes were injected with a sperm labeled with a mitochondrion-specific vital dye to help identify the contribution of the sperm to zygotic structures, in particular the centrosome. Oocytes were fixed at set intervals after sperm injection and examined by confocal laser scanning microscopy. In unfertilized oocytes, microtubules were present only in the metaphase-arrested second meiotic spindle and the first polar body. After sperm injection, an aster of microtubules formed adjacent to the sperm head and subsequently enlarged such that at the time of pronucleus migration and apposition it filled the entire cytoplasm. During syngamy, the microtubule matrix reorganized to form a mitotic spindle on which the chromatin of both parents aligned. Finally, after nuclear and cellular cleavage were complete, the microtubule asters dispersed into the interphase daughter cells. Sham injection induced parthenogenetic activation of 76% of oocytes, marked by the formation of multiple cytoplasmic microtubular foci that later developed into a dense microtubule network surrounding the female pronucleus. The finding that a parthenote alone can produce a microtubule aster, whereas the aster invariably forms at the base of the sperm head during normal fertilization, indicates that both gametes contribute to the formation of the zygotic centrosome in the horse. Finally, 25% of sperm-injected oocytes failed to complete fertilization, mostly due to absence of oocyte activation (65%), which was often accompanied by failure of sperm decondensation. In conclusion, this study demonstrated that union of the parental genomes in horse zygotes is accompanied by a series of integrated cytoskeleton-mediated events, failure of which results in developmental arrest.
DOI：
10.1073/pnas.82.12.4152
发表时间：
1985-01-01
影响因子：
11.1
作者：
SCHATTEN, G;SIMERLY, C;SCHATTEN, H
通讯作者：
SCHATTEN, H
影响因子：
3.6
作者：
Sutovsky, P;Navara, CS;Schatten, G
通讯作者：
Schatten, G