Differential control of activin, inhibin and follistatin proteins in cultured rat granulosa cells.
Differential control of activin, inhibin and follistatin proteins in cultured rat granulosa cells.
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培养的大鼠颗粒细胞中激活素、抑制素和卵泡抑素蛋白的差异控制。
DOI：
10.1006/bbrc.1993.1812
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发表时间：
1993
影响因子：
3.1
通讯作者：
Shimasaki,S
中科院分区：
文献类型：
--
作者：
Miyanaga,K;Erickson,GF;DePaolo,LV;Ling,N;Shimasaki,S
卵泡抑素、激活素和卵泡抑素蛋白由颗粒细胞产生，但控制其产生的机制尚不清楚。在这里，我们研究了蛋白激酶A（PKA）和蛋白激酶C（PKC）途径如何作用和相互作用，以调节这些蛋白质的产生。将未成熟大鼠的颗粒细胞与PKA通路激活剂（100 ng/ml FSH，10 μM毛喉素）和/或PKC通路激活剂（100 nM GnRH激动剂，100 nM 2-0-十四烷酰基-佛波醇-13-乙酸酯，TPA）一起培养。用重组人125 I-卵泡抑素通过配体印迹法测定条件培养基中的卵泡抑素和卵泡抑素，用冷激活素加125 I-卵泡抑素通过双配体印迹法测定条件培养基中的卵泡抑素。FSH和毛喉素刺激了卵泡刺激素而不是激活素的产生。相反，GnRH和TPA刺激激活素，并在较小程度上，激活素的生产;显着，这是第一次证明激活素二聚体生产的颗粒细胞。PKA通路的激活剂拮抗PKC效应物的作用，反之亦然。所有药物都增加了卵泡抑素蛋白的产生，PKA和PKC激活剂相互作用，进一步增加了卵泡抑素的产生。这些结果表明，FSH-PKA信号通路有利于αβ受体二聚体的形成，而GnRH-PKC通路有利于β亚基激活素二聚体的形成。这两种途径都能增加卵泡抑素蛋白的产生。
Follistatin, activin and inhibin proteins are produced by granulosa cells, but the mechanisms controlling their production remain unclear. Here, we examined how the protein kinase A (PKA) and protein kinase C (PKC) pathways act and interact to regulate production of these proteins. Granulosa cells from immature rats were cultured with activators of the PKA pathway (100 ng/ml FSH, 10 μM forskolin) and/or activators of the PKC pathway (100 nM GnRH agonist, 100nM 2-0-tetradecanoyl-phorbol-13-acetate, TPA). Conditioned media were assayed for inhibin and activin by ligand blotting using recombinant human125I-follistatin and for follistatin by double ligand blotting using cold activin plus125I-follistatin. FSH and forskolin stimulated inhibin but not activin production. In contrast, GnRH and TPA stimulated activin, and to a lesser degree, inhibin production; significantly, this is the first demonstration of activin dimer production by granulosa cells. Activators of the PKA pathway antagonized the actions of PKC effectors and vice versa. All agents increased follistatin protein production, and the PKA and PKC activators interacted to generate further increases in follistatin production. These results show that the FSH-PKA signalling pathway favors formation of αβ inhibin dimers while the GnRH-PKC pathway favors formation of β-subunit activin dimers. Both pathways act to increase follistatin protein production.