人类囊胚在与母亲年龄和染色体组成相关方面表现出独特的微小RNA谱。
Human blastocysts exhibit unique microrna profiles in relation to maternal age and chromosome constitution.
作者信息
McCallie Blair R, Parks Jason C, Strieby Alyssa L, Schoolcraft William B, Katz-Jaffe Mandy G
机构信息
National Foundation for Fertility Research, 10290 RidgeGate Circle, Lone Tree, CO, 80124, USA.
出版信息
J Assist Reprod Genet. 2014 Jul;31(7):913-9. doi: 10.1007/s10815-014-0235-y. Epub 2014 Apr 24.
PURPOSE
To determine microRNA (miRNA) expression in human blastocysts relative to advanced maternal age and chromosome constitution.
METHODS
Cryopreserved human blastocysts were warmed and underwent a trophectoderm biopsy for comprehensive chromosomal screening. Select blastocysts were then lysed, reverse transcribed, and pre-amplified prior to running real-time PCR. Statistical analysis was performed using an internal constant housekeeping miRNA. Significant microRNA's of interest were then analyzed for their predicted genes and biological pathways. Additional cryopreserved blastocysts were warmed and stained for the SIRT1 protein for validation.
RESULTS
Human blastocysts exhibit unique miRNA expression profiles in relation to maternal age and chromosome constitution. miR-93 was exclusively expressed in blastocysts from women in their forties and further up-regulated with an abnormal chromosome complement. Up-regulated miR-93 resulted in an inverse down-regulation of targets like SIRT1, resulting in reduced oxidative defense.
CONCLUSIONS
MiRNAs play an important role in aging as well as chromosome constitution and have downstream effects that regulate proteins which can compromise embryonic development.
目的
确定人类囊胚中微小RNA（miRNA）的表达与高龄产妇及染色体构成的关系。
方法
将冷冻保存的人类囊胚解冻，并进行滋养外胚层活检以进行全面的染色体筛查。然后选择囊胚进行裂解、逆转录，并在进行实时聚合酶链反应之前进行预扩增。使用内部恒定的管家miRNA进行统计分析。然后分析感兴趣的重要微小RNA的预测基因和生物学途径。将额外的冷冻保存囊胚解冻并进行SIRT1蛋白染色以进行验证。
结果
人类囊胚在与产妇年龄和染色体构成相关方面表现出独特的miRNA表达谱。miR-93仅在四十多岁女性的囊胚中表达，并随着染色体组异常而进一步上调。miR-93上调导致SIRT1等靶标的反向下调，从而降低氧化防御能力。
结论
微小RNA在衰老以及染色体构成中起重要作用，并具有调节可能损害胚胎发育的蛋白质的下游效应。
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