Lack of effect of cyclic adenosine monophosphate modulation during prematuration in vitro on development to the blastocyst stage and expression of specific genes associated with lineage commitment

Raquel Desenzi , Camila J. Cuellar , Quinn A. Hoorn , André Mariano Batista , Rafael Artur da Silva Jr. , Peter J. Hansen
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Abstract

Pharmacological elevation of cyclic AMP (cAMP) of cultured cumulus-oocyte complexes (COC) before or coincident with initiation of maturation has been reported to improve outcomes for various systems for in vitro production of embryos. Here it was hypothesized that artificial elevation of cAMP in the oocyte for a 2-h period of prematuration would improve developmental competence of matured oocytes and result in increased blastocyst yield and altered expression of genes important for embryonic differentiation. Treated COC were cultured for 2 h with dibutyryl cAMP (dbcAMP), a membrane-permeable form of cAMP, and 3-isobutyl-1-methylxanthine (IBMX), which inhibits phosphodiesterases that convert cAMP to ATP. Subsequently, oocytes were matured and fertilized and the resultant embryos cultured for 7.5 d. There was no effect of treatment on the percent of oocytes or cleaved embryos becoming blastocysts or on transcript abundance of EOMES, GATA4, NANOG, SOX2, or SOX17 in the blastocyst. Results do not support the use of pharmacological enhancers of cAMP concentrations in the oocyte for improving the blastocyst yield following in vitro oocyte maturation, fertilization, and embryo culture.
体外早熟期环腺苷单磷酸调节对囊胚发育和与谱系承诺相关的特定基因表达的影响缺乏。
据报道,在成熟开始之前或同时,培养的卵丘-卵母细胞复合物(COC)的环AMP (cAMP)的药理学升高可以改善各种体外胚胎生产系统的结果。本研究假设,在成熟前2小时人工升高卵母细胞中的cAMP可以提高成熟卵母细胞的发育能力,增加囊胚产量,改变胚胎分化重要基因的表达。处理后的COC与二丁基cAMP (dbcAMP)(一种膜渗透形式的cAMP)和3-异丁基-1-甲基黄嘌呤(IBMX)一起培养2小时,IBMX抑制将cAMP转化为ATP的磷酸二酯酶。随后,卵母细胞成熟并受精,形成的胚胎培养7.5 d。处理对卵母细胞或卵裂胚胎成为囊胚的百分比以及囊胚中EOMES、GATA4、NANOG、SOX2或SOX17的转录物丰度没有影响。结果不支持在体外卵母细胞成熟、受精和胚胎培养后,使用药物增强剂提高卵母细胞cAMP浓度来提高囊胚产量。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
JDS communications
JDS communications Animal Science and Zoology
CiteScore
2.00
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