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KPNA7是一种卵母细胞和胚胎特异性核转运蛋白α亚型,是猪胚胎发育所必需的。

KPNA7, an oocyte- and embryo-specific karyopherin α subtype, is required for porcine embryo development.

作者信息

Wang Xin, Park Ki-Eun, Koser Stephanie, Liu Shihong, Magnani Luca, Cabot Ryan A

机构信息

Department of Animal Sciences, Purdue University, 915 West State Street, West Lafayette, IN 47907, USA.

出版信息

Reprod Fertil Dev. 2012;24(2):382-91. doi: 10.1071/RD11119.

Abstract

Coordinated partitioning of intracellular cargoes between nuclear and cytoplasmic compartments is critical for cell survival and differentiation. The karyopherin α/β heterodimer functions to import cytoplasmic proteins that possess classical nuclear localisation signals into the nucleus. Seven karyopherinαsubtypes have been identified in mammals. The aim of this study was to determine the relative abundance of transcripts encoding seven karyopherinαsubtypes in porcine oocytes and embryos at discrete stages of cleavage development, and to determine the developmental requirements of karypopherinα7 (KPNA7), an oocyte and cleavage stage embryo-specific karyopherinαsubtype. We hypothesised that knockdown of KPNA7 would negatively affect porcine cleavage development. To test this hypothesis, in vitro matured and fertilised porcine oocytes were injected with a double-stranded interfering RNA molecule that targeted KPNA7; nuclei were counted in all embryos 6 days after fertilisation. Embryos injected with KPNA7-interfering RNAs possessed significantly lower cell numbers than their respective control groups (P<0.05). In vitro binding assays also suggest that KPNA7 may transport intracellular proteins that possess unique nuclear localisation signals. Our data suggest that embryos have differential requirements for individual karyopherinαsubtypes and that these karyopherinαsubtypes differentially transport intracellular cargo during cleavage development.

摘要

细胞内货物在细胞核和细胞质区室之间的协调分配对于细胞存活和分化至关重要。核转运蛋白α/β异二聚体的功能是将具有经典核定位信号的细胞质蛋白导入细胞核。在哺乳动物中已鉴定出七种核转运蛋白α亚型。本研究的目的是确定在猪卵母细胞和胚胎发育的不同卵裂阶段,编码七种核转运蛋白α亚型的转录本的相对丰度,并确定卵母细胞和卵裂期胚胎特异性核转运蛋白α亚型——核转运蛋白α7(KPNA7)的发育需求。我们假设敲低KPNA7会对猪的卵裂发育产生负面影响。为了验证这一假设,向体外成熟和受精的猪卵母细胞中注射靶向KPNA7的双链干扰RNA分子;在受精6天后对所有胚胎中的细胞核进行计数。注射了KPNA7干扰RNA的胚胎的细胞数量明显低于各自的对照组(P<0.05)。体外结合试验还表明,KPNA7可能转运具有独特核定位信号的细胞内蛋白。我们的数据表明,胚胎对单个核转运蛋白α亚型有不同的需求,并且这些核转运蛋白α亚型在卵裂发育过程中差异地转运细胞内货物。

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