AMP-activated protein kinase is involved in hormone-induced mouse oocyte meiotic maturation in vitro
AMP-activated protein kinase is involved in hormone-induced mouse oocyte meiotic maturation in vitro
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DOI：
10.1016/j.ydbio.2007.09.043
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发表时间：
2008-01-01
影响因子：
2.7
通讯作者：
Downs, Stephen M.
中科院分区：
文献类型：
--
作者：
Chen, Jing;Downs, Stephen M.
我们以前已经证明，AMP激活的蛋白激酶（AMPK）可以诱导小鼠卵母细胞在体外减数分裂停滞中恢复减数分裂。本研究的目的是确定AMPK激活是否参与胚胎诱导的成熟。卵泡刺激素（FSH）和EGF样肽，双调蛋白（AR），是卵丘细胞封闭的卵母细胞（CEO）成熟的有效诱导剂。FSH处理3小时内，磷酸乙酰辅酶A羧化酶（ACC）水平增加，在生发囊泡（GV）阶段的卵母细胞相比，非刺激的控制，并保持在整个9小时的培养升高，表明AMPK激活。培养6 h后观察到对AR的类似反应。使用抗PT 172抗体（仅结合活化的AMPK），Western分析表明，在FSH或AR处理的GV期卵母细胞中，AMPK活性在6 lt内。AMPK抑制剂，化合物C和腺嘌呤9-β-D-阿拉伯呋喃糖苷（araA），阻断FSH或AR诱导的减数分裂恢复和ACC磷酸化，进一步支持AMPK在FSH诱导的减数分裂恢复中的因果作用。使用抗-PT 172-AMPK抗体的免疫细胞化学显示，在用AMPK激活剂5-氨基咪唑-4-甲酰胺-1-β-D-呋喃核糖苷（AICAR）或用FSH或AR处理后，卵母细胞的生殖囊泡中弥散性细胞质染色增加，点状染色更强烈，并且这种染色被化合物C或抗-PT 172抗体的封闭肽消除。染色的卵母细胞从hCG刺激的小鼠与抗-PT 172抗体也表现出显着的标记在1-2 IT内的生殖囊泡。此外，在所有组的卵母细胞中，活跃的AMPK总是观察到与凝聚的染色体成熟的卵母细胞。总之，这些结果支持AMPK在FSH和AR诱导的体外成熟和hCG诱导的体内成熟中的作用。(c)2007年爱思唯尔公司All rights reserved.
We have previously shown that AMP-activated protein kinase (AMPK) can induce the resumption of meiosis in mouse oocytes maintained in meiotic arrest in vitro. The present study was carried out to determine whether AMPK activation is involved in hormone-induced maturation. Follicle-stimulating hormone (FSH) and the EGF-like peptide, amphiregulin (AR), are potent inducers of maturation in cumulus cell-enclosed oocytes (CEO). Within 3 h of FSH treatment, phospho-acetyl CoA carboxylase (ACC) levels were increased in germinal vesicle (GV)-stage oocytes when compared to non-stimulated controls and remained elevated throughout 9 h of culture, indicating AMPK activation. A similar response to AR was observed after 6 h of culture. Using anti-PT172 antibody (binds only to activated AMPK), Western analysis demonstrated active AMPK in both FSH- or AR-treated GV-stage oocytes within 6 It. The AMPK inhibitors, compound C and adenine 9-beta-D-arabinofuranoside (araA), blocked FSH- or AR-induced meiotic resumption and ACC phosphorylation, further supporting a causal role for AMPK in hormone-induced meiotic resumption. Immunocytochemistry using anti-PT172-AMPK antibody showed an increased diffuse cytoplasmic staining and more intense punctate staining in the germinal vesicles of oocytes following treatment with the AMPK activator 5-aminoimidazole-4carboxamide-1-beta-D-ribofuranoside (AICAR) or with FSH or AR, and this staining was eliminated by compound C or a blocking peptide for the anti-PT172 antibody. Staining of oocytes from hCG-stimulated mice with the anti-PT172 antibody also showed pronounced label in the germinal vesicles within 1-2 It. Furthermore, in oocytes from all groups, active AMPK was always observed in association with the condensed chromosomes of maturing oocytes. Taken together, these results support a role for AMPK in FSH and AR-induced maturation in vitro and hCG-induced maturation in vivo. (c) 2007 Elsevier Inc. All rights reserved.