Expression profile of genes as indicators of developmental competence and quality of in vitro fertilization and somatic cell nuclear transfer bovine embryos.
Expression profile of genes as indicators of developmental competence and quality of in vitro fertilization and somatic cell nuclear transfer bovine embryos.
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DOI：
10.1371/journal.pone.0108139
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Mutto A
中科院分区：
文献类型：
--
作者：
Cánepa MJ;Ortega NM;Monteleone MC;Mucci N;Kaiser GG;Brocco M;Mutto A
体外受精（IVF）和体细胞核移植（SCNT）等生殖生物技术提高了动物的生殖效率。然而，体外胚胎的出生率仍然落后于体内胚胎的出生率。因此，开发一套准确的胚胎能力评估和预测系统，无论是用于商业目的还是用于科学研究，都至关重要。先前的研究表明，体外培养系统诱导不同转录本的相对丰度（RA）发生变化，从而影响胚胎质量。本研究的目的是分析体外受精或SCNT产生的牛囊胚中涉及细胞应激（热休克蛋白70-kDa， HSP70）、内质网应激（免疫球蛋白重链结合蛋白，Bip，蛋白酶体亚基β5， PSMB5）和凋亡（BCL-2相关X蛋白，Bax，半胱氨酸天冬氨酸蛋白酶-3，Caspase-3）的一组基因的RA，并将其与体内对应基因进行比较。Poly (A) + mRNA从每组10个囊胚的3个池中分离，并通过实时RT-PCR进行分析。与体内源性囊胚相比，SCNT胚胎中三个应激指标（Bax、PSMB5和Bip）的RA显著升高。HSP70和Caspase-3基因转录本的RA差异无统计学意义。本研究可作为形态学分析的补充，为胚胎质量诊断提供一种有效准确的技术，最终有助于提高辅助生殖技术（ART）的效率。
Reproductive biotechnologies such as in vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT) enable improved reproductive efficiency of animals. However, the birth rate of in vitro-derived embryos still lags behind that of their in vivo counterparts. Thus, it is critical to develop an accurate evaluation and prediction system of embryo competence, both for commercial purposes and for scientific research. Previous works have demonstrated that in vitro culture systems induce alterations in the relative abundance (RA) of diverse transcripts and thus compromise embryo quality. The aim of this work was to analyze the RA of a set of genes involved in cellular stress (heat shock protein 70-kDa, HSP70), endoplasmic reticulum (ER) stress (immunoglobulin heavy chain binding protein, Bip; proteasome subunit β5, PSMB5) and apoptosis (BCL-2 associated X protein, Bax; cysteine aspartate protease-3, Caspase-3) in bovine blastocysts produced by IVF or SCNT and compare it with that of their in vivo counterparts. Poly (A) + mRNA was isolated from three pools of 10 blastocysts per treatment and analyzed by real-time RT-PCR. The RA of three of the stress indicators analyzed (Bax, PSMB5 and Bip) was significantly increased in SCNT embryos as compared with that of in vivo-derived blastocysts. No significant differences were found in the RA of HSP70 and Caspase-3 gene transcripts. This study could potentially complement morphological analyses in the development of an effective and accurate technique for the diagnosis of embryo quality, ultimately aiding to improve the efficiency of assisted reproductive techniques (ART).
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