Paracrine and autocrine regulation of epidermal growth factor-like factors in cumulus oocyte complexes and granulosa cells: Key roles for prostaglandin synthase 2 and progesterone receptor
Paracrine and autocrine regulation of epidermal growth factor-like factors in cumulus oocyte complexes and granulosa cells: Key roles for prostaglandin synthase 2 and progesterone receptor
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DOI：
10.1210/me.2005-0504
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发表时间：
2006-06-01
影响因子：
--
通讯作者：
Richards, JoAnne S.
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文献类型：
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作者：
Shimada, Masayuki;Hernandez-Gonzalez, Inmaculada;Richards, JoAnne S.
将促黄体生成素（LH）峰与几乎没有LH受体的卵丘细胞功能变化相联系的分子桥梁正在被揭示。在此我们证明，表皮生长因子（EGF）样因子双调蛋白（Areg）、上皮调节蛋白（Ereg）和β细胞素（Btc）通过涉及前列腺素（PGs）和孕激素受体（PGR）作用的自分泌和旁分泌机制在卵丘卵母细胞复合体（COCs）中被诱导。在人绒毛膜促性腺激素处理后4小时和8小时，分别从前列腺素合酶2（Ptgs2）基因敲除小鼠和孕激素受体基因敲除（PRKO）小鼠收集的COCs和卵巢中，Areg和Ereg的mRNA及蛋白质水平显著降低。在培养的COCs中，促卵泡激素（FSH）/毛喉素在0.5小时内诱导Areg mRNA，在4小时达到峰值，这一过程被p38丝裂原活化蛋白激酶（MAPK）抑制剂（SB203580）、MAPK激酶（MEK）1抑制剂（PD98059）和PTGS2抑制剂（NS398）阻断，但不被蛋白激酶A（PKA）抑制剂（KT5720）阻断。相反，AREG而非FSH在0.5小时诱导Ptsg2 mRNA，Ptgs2和Areg mRNA在4小时表达达到峰值，这些过程被表皮生长因子受体酪氨酸激酶抑制剂AG1478（AG）、PD98059和NS398阻断。PGE2逆转了AG对AREG诱导的Areg表达的抑制作用，但对Ptgs2无此作用，表明Ptgs2位于表皮生长因子受体（EGF - R）信号通路的下游。佛波醇12 - 肉豆蔻酸13 - 乙酸酯（PMA）和腺病毒表达的PGRA在颗粒细胞中协同诱导Areg mRNA。在COCs中，AREG不仅诱导影响基质形成的基因，还诱导参与类固醇生成（StAR，Cyp11a1）和免疫细胞样功能（Pdcd1，Runx1，Cd52）的基因。总之，FSH通过p38MAPK介导的Areg mRNA诱导先于AREG通过ERK1/2诱导的Ptgs2 mRNA。前列腺素通过卵丘细胞中的PTGER2发挥作用，为调节COCs中Areg的表达提供了一条次要的自分泌途径，显示了排卵卵泡中G蛋白偶联受体和生长因子受体通路之间的关键功能联系。
The molecular bridges that link the LH surge with functional changes in cumulus cells that possess few LH receptors are being unraveled. Herein we document that epidermal growth factor ( EGF)- like factors amphiregulin ( Areg), epiregulin ( Ereg), and betacellulin ( Btc) are induced in cumulus oocyte complexes ( COCs) by autocrine and paracrine mechanisms that involve the actions of prostaglandins ( PGs) and progesterone receptor ( PGR). Areg and Ereg mRNA and protein levels were reduced significantly in COCs and ovaries collected from prostaglandin synthase 2 ( Ptgs2) null mice and Pgr null ( PRKO) mice at 4 h and 8 h after human chorionic gonadotropin, respectively. In cultured COCs, FSH/forskolin induced Areg mRNA within 0.5 h that peaked at 4 h, a process blocked by inhibitors of p38MAPK ( SB203580), MAPK kinase (MEK) 1 ( PD98059), and PTGS2 ( NS398) but not protein kinase A ( PKA) ( KT5720). Conversely, AREG but not FSH induced Ptsg2 mRNA at 0.5 h with peak expression of Ptgs2 and Areg mRNAs at 4 h, processes blocked by the EGF receptor tyrosine kinase inhibitor AG1478 ( AG), PD98059, and NS398. PGE2 reversed the inhibitory effects of AG on AREG-induced expression of Areg but not Ptgs2, placing Ptgs2 downstream of EGF-R signaling. Phorbol 12- myristate 13-acetate (PMA) and adenovirally expressed PGRA synergistically induced Areg mRNA in granulosa cells. In COCs, AREG not only induced genes that impact matrix formation but also genes involved in steroidogenesis ( StAR, Cyp11a1) and immune cell-like functions (Pdcd1, Runx1, Cd52). Collectively, FSH-mediated induction of Areg mRNA via p38MAPK precedes AREG induction of Ptgs2 mRNA via ERK1/2. PGs acting via PTGER2 in cumulus cells provide a secondary, autocrine pathway to regulate expression of Areg in COCs showing critical functional links between G protein- coupled receptor and growth factor receptor pathways in ovulating follicles.