Prostaglandin-D synthetase induces transcription of the LH beta subunit in the primary culture of chicken anterior pituitary cells via the PPAR signaling pathway
Prostaglandin-D synthetase induces transcription of the LH beta subunit in the primary culture of chicken anterior pituitary cells via the PPAR signaling pathway
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DOI：
10.1016/j.theriogenology.2009.09.020
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发表时间：
2010-02-01
期刊：
影响因子：
2.8
通讯作者：
Shiue, Y. -L.
中科院分区：
文献类型：
--
作者：
Chen, L. -R.;Lee, S. -C.;Shiue, Y. -L.
本研究以鸡垂体前叶细胞为研究对象，研究了鸡垂体素D合成酶（PGDS）对鸡产蛋的调控作用。PGDS下游代谢物PGD（2）或PGJ（2）均以剂量和时间依赖性方式升高LHB mRNA和LHB蛋白水平，单独用花生四烯酸（1 μ M）处理可上调15-脱氧-Delta（12，14）-PGJ（2）（15-d-PGJ（2）;来源于PGJ（2））/PGJ（2）、LHB mRNA和LHB蛋白水平在原代培养的鸡垂体前叶细胞中显著升高（P < 0.05）。将质粒增强的绿色荧光蛋白-PGDS质粒转染到含有1 μ M花生四烯酸的培养基中的这些细胞中，另外增加了15-d-PGJ（2）/PGJ（2）、LHB mRNA和LHB蛋白水平（P < 0.05）。在产蛋母鸡下丘脑/垂体中，产蛋量高的L2台湾土鸡PGDS与PPARA mRNA水平高度相关（r = 0.64; P < 0.05）。商品单梳白色来航蛋鸡PGDS与PPARA、PPARG mRNA水平之间均存在高度相关性（r = 0.65; P < 0.05; r = 0.67; P < 0.01）。一个广泛的PPARs激动剂，GW 9578（5至500 nM），提高LHB mRNA和LHB蛋白水平（P < 0.05）。PPARA特异性（GW 6471）和PPARG特异性（T0070907）拮抗剂抑制内源性LHB mRNA和LHB蛋白水平（P < 0.05）;此外，两种拮抗剂均减弱花生四烯酸诱导的LHB mRNA水平，（P < 0.05）和PGDS诱导的（1 β，M花生四烯酸存在下）鸡垂体前叶细胞LHB mRNA和LHB蛋白水平（P < 0.05）。在PGD（2-）、PGJ（2-）、花生四烯酸、PGDS和GW 9578诱导的以及GW 6471和T0070907抑制的垂体前叶细胞中，LHB mRNA/LHB蛋白比例较高，表明LHB转录发生在翻译之前。总之，PGDS通过PPAR信号通路诱导LHB转录和随后的翻译。(C)2010年爱思唯尔公司All rights reserved.
Downstream effects of prostaglandin-D synthetase (PGDS) in a primary culture of chicken (Gallus gallus) anterior pituitary cells were investigated to study how PGDS regulated laying in hens. Either PGDS downstream metabolite, PGD(2) or PGJ(2), elevated LHB mRNA and LHB protein levels in dose- and time-dependent manners, and treatment with arachidonic acid (1 mu M) alone upregulated 15-deoxy-Delta(12,14)-PGJ(2) (15-d-PGJ(2); derived from PGJ(2))/PGJ(2), LHB mRNA, and LHB protein levels (P < 0.05) in the primary culture of chicken pituitary anterior cells. Transfection of the plasmid Enhanced Green Fluorescent Protein-PGDS plasmid into these cells in medium containing 1 mu M arachidonic acid additionally increased 15-d-PGJ(2)/PGJ(2), LHB mRNA, and LHB protein levels (P < 0.05). In the hypothalamus/pituitary gland of laying hens, there was a high correlation (r = 0.64; P < 0.05) between PGDS and the peroxisome proliferator-activated receptor A (PPARA) mRNA level in a high egg production strain, the L2 Taiwan country chickens. In commercial Single-Comb White Leghorn layers, there were high correlation coefficients between PGDS and PPARA (r = 0.65; P < 0.05) and between PGDS and PPARG (r = 0.67; P < 0.01) mRNA levels. A broad-range PPARs agonist, GW9578 (5 to 500 nM), enhanced LHB mRNA and LHB protein levels (P < 0.05). The PPARA-specific (GW6471) and PPARG-specific (T0070907) antagonists suppressed endogenous LHB mRNA and LHB protein levels (P < 0.05); in addition, both antagonists attenuated arachidonic acid induced LHB mRNA levels (P < 0.05) and PGDS-induced (in the presence of 1 p,M arachidonic acid) LHB mRNA and LHB protein (P < 0.05) levels in the primary culture of chicken anterior pituitary cells. Higher LHB mRNA/LHB protein ratios in PGD(2-), PGJ(2-), arachidonic acid-, PGDS-, and GW9578-induced as well as GW6471- and T0070907-suppressed anterior pituitary cells suggested that LHB transcription occurred before translation. In conclusion, PGDS induced LHB transcription and subsequent translation via the PPAR signaling pathway. (C) 2010 Elsevier Inc. All rights reserved.