Effects of environmental estrogenic pollutants on cellular functions in the noradrenergic neurons and search for their receptors

环境雌激素污染物对去甲肾上腺素能神经元细胞功能的影响及其受体的寻找

基本信息

批准号：
17590111
负责人：
TAKAHASHI Kojiro
金额：
$ 2.24万
依托单位：
University of Occupational and Environmental Health
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

项目摘要

Environmental estrogenic pollutants are compounds that have been shown to have estrogenic effects on fetal development and reproductive systems. Less attention, however, has been paid to their influence on neuronal functions. We have examined the effects of estrogenic pollutants on catecholamine synthesis in adrenal medullary cells used as a model system of noradrenergic neurons. Treatment of cultured bovine adrenal medullary cells with p-nonylphenol and bisphenol A at 10 nM for 3 days stimulated ^<14>C-catecholamine synthesis from [^<14>C]tyrosine and tyrosine hydroxylase activity, an effect that was not inhibited by ICI 182,780, an inhibitor of estrogen receptors. Significant effects of p-nonylphenol on ^<14>C-catecholamine synthesis were observed at 0.1 nM that is 50 times lower than that of the international regulatory standard (5 nM) and the maximum effects were around at 10 nM. Short-term treatment of cells with 10 nM p-nonylphenol for 10 min also activated tyrosine hydroxylase, … More which was suppressed by U0126, an inhibitor of mitogen-activated protein kinase (MAPK) kinase. Furthermore, treatment of cells with p-nonylphenol for 5 min increased the phospho-p44/42MAPK in a concentration (1 - 1000 nM)-dependent manner.Incubation of adrenal medullary cells with 17β-estradiol(E_2) (0.3-100 nM) or membrane-impermeable E_2-bovine serum albumin (100 nM) also acutely increased ^<14>C-catecholamine synthesis from [^<14>C]tyrosine. The plasma membrane isolated from the gland showed two classes of specific binding sites of [^3H] E_2 with apparent Kds of 3.2 and 106 nM and Bmaxs of 0.44 and 8.5 pmol/mg protein, respectively. The high-affinity binding of [^3H] E_2 was most strongly inhibited by E_2 and phytoestrogens, and to lesser extents by other steroid hormones, while it was enhanced by ICI182,780 and environmental estrogenic pollutants.These findings suggest that E_2 and environmental estrogenic pollutants acutely stimulates catecholamine synthesis via activation of p44/42MAPK through unique estrogen receptors in the plasma membrane of bovine adrenal medulla. Less

环境雌激素污染物是已被证明对胎儿发育和生殖系统具有雌激素作用的化合物，然而，我们研究了雌激素污染物对肾上腺髓质细胞中儿茶酚胺合成的影响。用作去甲肾上腺素能神经元的模型系统，用 10 nM 对壬基酚和双酚 A 处理培养的牛肾上腺髓质细胞 3 天。刺激由[^ 14 C]酪氨酸和酪氨酸羟化酶活性合成^ 14 C-儿茶酚胺，该作用不受ICI 182,780(雌激素受体抑制剂)的抑制。对壬基酚对^ 14 C有显着影响。观察到 C-儿茶酚胺合成浓度为 0.1 nM，比国际监管标准 (5 nM) 低 50 倍，最大效应约为 10 nM。用 10 nM 对壬基苯酚短期处理细胞 10 分钟也会激活酪氨酸羟化酶，而该酶被丝裂原激活蛋白激酶 (MAPK) 激酶抑制剂 U0126 抑制。此外，用对壬基酚处理细胞 5 分钟，磷酸化 p44/42MAPK 的浓度增加了 (1 - 1000 nM）依赖性方式。用17β-雌二醇（E_2）（0.3-100 nM）或膜不可渗透的E_2-牛血清白蛋白（100 nM）孵育肾上腺髓质细胞也急剧增加[14] C-儿茶酚胺的合成。从腺体分离的质膜显示出两类[^ 3H]特异性结合位点。 E_2 的表观 Kd 分别为 3.2 和 106 nM，Bmax 分别为 0.44 和 8.5 pmol/mg 蛋白质 [^3H] E_2 的高亲和力结合最受 E_2 和植物雌激素的抑制，其他类固醇激素的抑制程度较小。，而 ICI182,780 和环境雌激素污染物则增强了它。这些发现表明 E_2 和环境雌激素污染物通过牛肾上腺髓质质膜中独特的雌激素受体激活 p44/42MAPK，急剧刺激儿茶酚胺合成。

项目成果

期刊论文数量（32）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Nephrogenic diabetes insipidus in mice lacking all nitric oxide synthase isoforms

DOI：
10.1073/pnas.0502236102
发表时间：
2005-07-26
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
影响因子：
11.1
作者：
Morishita, T;Tsutsui, M;Yanagihara, N
通讯作者：
Yanagihara, N

Effect of prolonged exposure to milnacipran on norepinephrine transporter in cultured bovine adrenal medullary cells

长期暴露于米那普仑对培养牛肾上腺髓质细胞去甲肾上腺素转运蛋白的影响