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非必需氨基酸在孤雌生殖或体细胞核移植产生的猪胚胎中的作用。

Role of nonessential amino acids on porcine embryos produced by parthenogenesis or somatic cell nuclear transfer.

作者信息

Gupta Mukesh Kumar, Uhm Sang Jun, Lee Sung-Ho, Lee Hoon Taek

机构信息

Department of Animal Biotechnology, Bio-Organ Research Center, Konkuk University, 1, Hwayang-dong, Gwangjin-Gu, Seoul, South Korea.

出版信息

Mol Reprod Dev. 2008 Apr;75(4):588-97. doi: 10.1002/mrd.20789.

Abstract

Amino acids play a multitude of roles during early embryonic development and have been demonstrated to facilitate improved development of in vivo or in vitro fertilized and parthenogenetic embryos in several species. However, review of emerging literatures, shows that culture milieu of cloned embryos might be different from those of in vitro fertilized embryos. This study therefore, evaluated the effect of nonessential amino acids (NEAA) on yield and quality of porcine embryos produced by somatic cell nuclear transfer (SCNT) and compared them with parthenogenetic embryos as control. Analysis showed that, supplementation of NEAA to culture medium significantly improved the blastocyst rate of parthenogenetic (38.9 +/- 8.8 vs. 27.5 +/- 9.0%) as well as SCNT (22.5 +/- 2.2 vs. 13.8 +/- 3.4%) embryos although cleavage rates were not different (P < 0.05). These blastocysts also had higher hatching ability and contained higher cell number than control blastocysts (P < 0.05). TUNEL labeling revealed that blastocysts cultured in the presence of NEAA were less predisposed to biochemical apoptosis and showed lower indices of TUNEL, fragmentation, and total apoptosis than those cultured in the absence of NEAA (P < 0.05). Real-time qRT-PCR analysis further revealed that NEAA decreased the expression ratio of BAX:BCL-xL and enhanced the relative abundance of IGF2 transcripts. Therefore, our study suggests that NEAA improves the yield and quality of cloned porcine embryos by enhancing blastocyst expansion, hatching, and total cell number and decreasing the apoptosis by positively modulating the expression of embryo survival related genes, similar to those reported for in vivo or in vitro fertilized embryos. Nonessential amino acids improve the yield and quality of cloned and parthenogenetic porcine embryos and modulate the expression of embryo survival related genes.

摘要

氨基酸在早期胚胎发育过程中发挥着多种作用,并且已证实在多个物种中,氨基酸有助于改善体内或体外受精胚胎以及孤雌生殖胚胎的发育。然而,对新兴文献的综述表明,克隆胚胎的培养环境可能与体外受精胚胎不同。因此,本研究评估了非必需氨基酸(NEAA)对体细胞核移植(SCNT)生产的猪胚胎产量和质量的影响,并将其与作为对照的孤雌生殖胚胎进行比较。分析表明,向培养基中添加NEAA可显著提高孤雌生殖胚胎(38.9±8.8%对27.5±9.0%)以及SCNT胚胎(22.5±2.2%对13.8±3.4%)的囊胚率,尽管卵裂率没有差异(P<0.05)。这些囊胚还具有更高的孵化能力,并且比对照囊胚含有更多的细胞数(P<0.05)。TUNEL标记显示,在NEAA存在下培养的囊胚比在无NEAA条件下培养的囊胚更不易发生生化凋亡,并且TUNEL、碎片化和总凋亡指数更低(P<0.05)。实时qRT-PCR分析进一步表明,NEAA降低了BAX:BCL-xL的表达比例,并提高了IGF2转录本的相对丰度。因此,我们的研究表明,NEAA通过增强囊胚扩张、孵化和总细胞数,并通过正向调节胚胎存活相关基因的表达来减少凋亡,从而提高克隆猪胚胎的产量和质量,这与体内或体外受精胚胎的报道相似。非必需氨基酸提高了克隆和孤雌生殖猪胚胎的产量和质量,并调节了胚胎存活相关基因的表达。

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