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首页> 外文学位 >Identification of intracellular calcium receptors (Inositol 1,4,5 -trisphosphate and ryanodine) during maturation in bovine oocytes.
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Identification of intracellular calcium receptors (Inositol 1,4,5 -trisphosphate and ryanodine) during maturation in bovine oocytes.

机译：鉴定牛卵母细胞成熟过程中的细胞内钙受体（肌醇1,4,5-三磷酸和ryanodine）。

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摘要

Intracellular calcium transients (oscillations) are critical to oocyte activation and subsequent development. Inositol 1,4,5-trisphosphate (IP3) receptor (IP3R) and ryanodine receptor (RyR) have been identified as two ligand-gated calcium channels that play a critical role in calcium release. Immature and mature bovine oocytes as well as fertilized eggs were used to evaluate the presence of the IP3R and the RyR as well as the spatial and temporal relationship of these receptors to endoplasmic reticulum (ER) and cortical granules (CG). Immunoprecipitation experiments were first carried out to verify the presence of these two receptors during early and late maturation using specific antibodies against these two receptors. Results indicate that IP3R is translated throughout the maturation period. In contrast, RyR is only translated during the late maturation period of bovine oocytes. The dynamic changes of RyR are due to the modification of protein synthesis rather than a conformational change during maturation of bovine oocytes.;Furthermore, immunocytochemistry, fluorescent staining, and laser scanning confocal microscopy were applied to reveal the localization of the IP3R and the RyR. The significant dynamics were observed in both IP3R and RyR at four different developmental stages from the germinal vesicle-intact stage (GV-intact) to 6 hours postinsemination (6 hpi). In addition, both the ER and the CG also showed apparent dynamic changes during meiotic maturation and at fertilization. Thus, based on the data, we proposed: (1) both IP3R and RyR contribute to the significant increase of intracellular calcium upon activation at fertilization, the dynamics of IP3R and RyR account for the increased intracellular calcium responsiveness during activation at fertilization; (2) the IP3R will associate with the ER, whereas the RyR will associate with the CG; (3) the IP3R and the RyR serve differently in calcium homeostasis and calcium release during meiotic maturation and at fertilization. In conclusion, both IP3R and RyR are present in bovine oocytes during maturation.
机译：细胞内钙瞬变（振荡）对于卵母细胞激活和随后的发育至关重要。肌醇1,4,5-三磷酸（IP3）受体（IP3R）和ryanodine受体（RyR）被确定为两个在钙释放中起关键作用的配体门控钙通道。使用未成熟和成熟的牛卵母细胞以及受精卵来评估IP3R和RyR的存在，以及这些受体与内质网（ER）和皮质颗粒（CG）的时空关系。首先使用针对这两种受体的特异性抗体，进行了免疫沉淀实验，以验证在早熟和晚熟过程中这两种受体的存在。结果表明，IP3R在整个成熟期内都进行了翻译。相反，RyR仅在牛卵母细胞成熟后期才翻译。 RyR的动态变化是由于蛋白质合成的修饰而不是牛卵母细胞成熟过程中的构象变化所致。此外，通过免疫细胞化学，荧光染色和激光扫描共聚焦显微镜观察了IP3R和RyR的定位。 IP3R和RyR在从生小泡完整期（GV完整）到授精后6小时（6 hpi）四个不同的发育阶段均观察到了显着的动力学。此外，ER和CG在减数分裂成熟和受精过程中也表现出明显的动态变化。因此，基于这些数据，我们提出：（1）IP3R和RyR都在受精激活时引起细胞内钙的显着增加，IP3R和RyR的动力学解释了受精激活时细胞内钙反应性的增加； （2）IP3R将与ER关联，而RyR将与CG关联； （3）IP3R和RyR在减数分裂成熟和受精过程中钙稳态和钙释放中的作用不同。总之，在成熟期间，牛卵母细胞中同时存在IP3R和RyR。

著录项

  • 作者

    Wang, Lin.;

  • 作者单位

    Utah State University.;

  • 授予单位 Utah State University.;
  • 学科 Biology Cell.;Agriculture Animal Pathology.
  • 学位 Ph.D.
  • 年度 2000
  • 页码 119 p.
  • 总页数 119
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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