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Profilin and actin-related proteins regulate microfilament dynamics during early mammalian embryogenesis

机译:蛋白酶蛋白和肌动蛋白相关蛋白在哺乳动物早期胚胎发生过程中调节微丝动力学

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摘要

BACKGROUND: Profilins are ubiquitous proteins widely distributed in animals, including humans. They regulate actin polymerization by sequestering actin monomers in association with other actin-related proteins (Arps). Actin remodelling is essential for oocyte maturation, fertilization and embryo development; yet the role of profilins in these events is not well understood. Here we investigate profilin distribution and function during bovine fertilization and early embryogenesis, and we examine profilin localization with respect to the co-distribution of other Arps. METHODS AND RESULTS: Western blotting, confocal microscopy with immunofluorescence and protein inhibition studies with antibodies were implemented. Profilin distributes inside interphase nuclei, throughout the cytoplasm and near the cell cortex at different stages of bovine oocyte maturation, fertilization and embryo development. Expression is detected through the blastocyst stage, where profilin localizes to the inner cell mass as well as trophectoderm. Profilin co-distributes with actin monomers and Arps vasodilator-stimulated phospho protein, p140mDia, Arp 3 and p80 coilin in pronucleate-stage zygotes. Antiprofilin antibodies inhibit normal embryo development by disrupting microfilaments, but not microtubules, and result in a higher concentration of profilin and p140mDia mislocalized to the cortex. CONCLUSIONS: These findings demonstrate that profilin regulates actin dynamics both within the cytoplasm and inside the nuclei of developing mammalian embryos and that its function is essential during fertilization to ensure successful development.
机译:背景:纤维蛋白原是广泛存在于动物(包括人类)中的蛋白质。它们通过螯合肌动蛋白单体与其他肌动蛋白相关蛋白(Arps)来调节肌动蛋白聚合。肌动蛋白重塑对于卵母细胞成熟,受精和胚胎发育至关重要。然而,对于这些事件中的profilins的作用还知之甚少。在这里,我们调查在牛受精和早期胚胎发生过程中profilin的分布和功能,并检查与其他Arps共分布有关的profilin定位。方法和结果:进行了蛋白质印迹,免疫荧光共聚焦显微镜和抗体的蛋白抑制研究。在牛卵母细胞成熟,受精和胚胎发育的不同阶段,Profilin分布在相间核内,整个细胞质和细胞皮质附近。通过胚泡阶段检测表达,在胚泡阶段,profilin定位于内部细胞团以及滋养外胚层。 Profilin与肌动蛋白单体和Arps血管舒张剂刺激的磷酸蛋白p140mDia,Arp 3和p80卷心素共分布在原核期受精卵中。抗profilin抗体通过破坏微丝而不是微管来抑制正常胚胎发育,并导致更高浓度的profilin和p140mDia错位到皮层。结论:这些发现表明,脯氨酸蛋白调节发育中的哺乳动物胚胎的细胞质内和细胞核内的肌动蛋白动力学,并且其功能在受精过程中对于确保成功发育至关重要。

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  • 来源
    《Human Reproduction》 |2006年第5期|1143-1153|共11页
  • 作者单位

    Centro de Estudios en Ginecología y Reproducción (CEGyR) Buenos Aires Argentina and;

    Pittsburgh Development Center Magee-Womens Research Institute Departments of Obstetrics Gynecology and Reproductive Sciences and Cell Biology and Physiology University of Pittsburgh School of Medicine Pittsburgh PA USA;

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