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首页> 外文期刊>Molecular reproduction and development >Effects of adenosine monophosphate-activated kinase activators on bovine oocyte nuclear maturation in vitro.
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Effects of adenosine monophosphate-activated kinase activators on bovine oocyte nuclear maturation in vitro.

机译：腺苷一磷酸激活的激酶激活剂对牛卵母细胞核成熟的影响。

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摘要

The purpose of this study was to examine the effects of an activator of AMPK (5-aminoimidazole-4-carboxamide 1-beta-D-ribofuranoside (AICAR)) on bovine oocyte nuclear maturation in vitro. After 7 hr of culture, AICAR (1 mM) significantly increased the percentages of cumulus-enclosed oocytes (CEO) and denuded oocytes (DO) remaining at the germinal vesicle stage. After 22 hr of culture, AICAR significantly reduced the percentage of CEO reaching metaphase II (MII). AICAR at 1.0 mM also increased the inhibitory effect of the adenylate cyclase activator forskolin in CEO; however, at 0.05 mM, AICAR increased the percentage of oocytes at MII after 22 hr of culture compared to forskolin alone. The adenosine kinase inhibitor 5'-aminodeoxyadenosine reversed the effect of AICAR in CEO and DO showing that phosphorylation of AICAR by adenosine kinase is required for its inhibitory activity. GMP, but not AMP, inhibited meiosis in CEO and DO; however, inhibition of guanyl and adenyl nucleotides synthesis did not reverse the effect of AICAR suggesting that the inhibitory effect of AICAR is not due to increased synthesis of these nucleotides. Metformin, another activator of AMPK, also inhibited GVBD in CEO and DO. The alpha-1 isoform of the catalytic subunit of AMPK was detected in oocytes and cumulus cells, and reverse transcription-polymerase chain reaction experiments showed the presence of transcripts for alpha-1, alpha-2, beta-1, and gamma-3 isoforms of the regulatory subunits in cumulus cells and oocytes. These data show that the AMPK activator AICAR is inhibitory to nuclear maturation in bovine oocytes due to activation of AMPK.
机译：这项研究的目的是检查AMPK激活剂（5-氨基咪唑-4-羧酰胺1-β-D-核呋喃糖苷（AICAR））对牛卵母细胞核成熟的影响。培养7小时后，AICAR（1 mM）显着增加了在生小泡阶段剩余的积卵卵母细胞（CEO）和裸露卵母细胞（DO）的百分比。培养22小时后，AICAR大大降低了CEO进入中期II（MII）的百分比。 AICAR浓度为1.0 mM时，还可以增强CEO中腺苷酸环化酶激活剂forskolin的抑制作用。然而，与单独的福司高林相比，在培养22小时后，AICAR在0.05 mM时增加了MII处卵母细胞的百分比。腺苷激酶抑制剂5'-氨基脱氧腺苷逆转了AICAR在CEO和DO中的作用，表明腺苷激酶对AICAR的磷酸化是其抑制活性所必需的。 GMP（而非AMP）抑制CEO和DO的减数分裂;然而，抑制胍基和腺苷酸核苷酸的合成并不能逆转AICAR的作用，这表明AICAR的抑制作用不是由于这些核苷酸的合成增加所致。二甲双胍，AMPK的另一种激活剂，也抑制CEO和DO中的GVBD。在卵母细胞和卵丘细胞中检测到AMPK催化亚基的α-1亚型，逆转录聚合酶链反应实验表明存在α-1，α-2，β-1和gamma-3亚型的转录本。卵丘细胞和卵母细胞中调节亚基的表达。这些数据表明，由于AMPK的活化，AMPK活化剂AICAR抑制了牛卵母细胞的核成熟。

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