Detection and quantification of maternal-effect gene transcripts in mouse second polar bodies: potential markers of embryo developmental competence.
Detection and quantification of maternal-effect gene transcripts in mouse second polar bodies: potential markers of embryo developmental competence.
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DOI：
10.1016/j.fertnstert.2013.02.003
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发表时间：
2013-06
影响因子：
6.7
通讯作者：
Woodruff TK
中科院分区：
文献类型：
--
作者：
Jiao ZX;Woodruff TK
为了验证第二极体(PB2)来源的mRNAs的量化为评估胚胎质量提供了一种非侵入性工具的假设。前瞻性研究医院学术研究实验室CD1雌性小鼠MII卵母细胞经孕马血清促性腺激素(PMSG)和人绒毛膜促性腺激素(HCG)启动后，从7至8周龄的小鼠获得。体外受精后，取受精卵的PB2进行逆转录。实时荧光定量聚合酶链式反应检测单个PB2的基因表达水平。将同胞受精卵连续培养至囊胚期。在所有候选基因中都检测到胚胎发育能力和6个母体效应基因(Dnmt1、Mater、Nobox、Npm2、Tcl1和Zar1)在PB2PB2mRNA中的转录本。受精卵中丰度较高的转录本更有可能在单个PB2的qPCR复制中被检测到。4个候选基因(Dnmt1、Nobox、Npm2和Tcl1)在两组(2-细胞胚胎和囊胚)PB2中的表达水平接近统计学意义。Pb2可能包含受精后受精卵的代表性转录谱。PB2基因表达的差异可能是胚胎质量的潜在生物标志物。
To test the hypothesis that quantification of mRNAs originating the second polar body (PB2) provides a non-invasive tool for assessing embryo quality. Prospective study Hospital-based academic research laboratory CD1 female mice MII oocytes obtained from 7- to 8-week-old mice after pregnant mare’s serum gonadotrophin (PMSG) and human chorionic gonadotrophin (hCG) priming. After in vitro fertilization, the PB2 was biopsied from zygotes followed by reverse transcription. Real-time PCR was performed to quantify gene expression levels in single PB2. The sibling zygotes were continuously cultured to blastocyst stage. Embryo developmental competence and six maternal-effect gene (Dnmt1, Mater, Nobox, Npm2, Tcl1 and Zar1) transcripts in the PB2 PB2 mRNA was detected in all candidate genes. Transcripts that were present in greater abundance in the zygote were more likely to be detected in qPCR replicates from single PB2. 4 candidate genes (Dnmt1, Nobox, Npm2 and Tcl1) expression levels in PB2 between two groups (2-cell embryo vs. blastocyts) approached statistical significance. PB2 may contain a representative transcript profile to that of the zygote after fertilization. Differences in gene expression in PB2 may be potential biomarkers of embryo quality.
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