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The sensitivity to endocrine disruptors and the establishment of sensitive bioassay for endocrine disruptors in poultry

家禽内分泌干扰物敏感性及内分泌干扰物敏感生物测定方法的建立

基本信息

批准号：
15580232
负责人：
SUGIYAMA Toshie
金额：
$ 2.3万
依托单位：
Niigata University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

1.Aromatic hydrocarbon receptor (AhR ; dioxin receptor) mRNA expression with aging.We detected the expression of AhR mRNA in various tissues of female chickens with aging (0 to 390 days old) by RT-PCR. As a result, AhR mRNA was detected in gizzard, liver, lung, kidney and heart during all stages. However, the oviduct and bone marrow of female chickens represented no AhR mRNA at 0-day stage and less at 30-90-day stages, respectively. These results suggest that the sensitivities of oviduct and bone marrow to dioxin are altered according to the age of female chickens.2.Expression and localization of AhR in oviduct of laying chickens.AhR expression was detected in magnum, isthmus and shell gland of oviducts by western blotting analysis. Especially in isthmus and shell gland, AhR was strongly localized in the cytoplasm of their epithelial cells. These results suggest that dioxin directly disrupts the metabolism of isthmus and shell gland, resulting in the eggshell weakness.3.In Vivo bioassa … More y of endocrine disruptors using avian medullary bone.The treatment of male Japanese quail with DDE (10mg/day) for 7 days induced medullary bone formation in marrow cavities of femurs. Concomitantly, the treatment increased the serum vitellogenin levels. These results suggest that avian medullary bone formation is superior as in vivo bioassay model of endocrine disruptors.4.Expression and localization of endocrine disrupter receptors in avian medullary bone.Estrogen receptor (α and β) and androgen receptor are well known as receptors for endocrine disruptor. In the present study by in situ hybridization and immunohistochemical analyses, both receptors (especially α subtype in estrogen receptor) were localized in osteoblasts of chicken medullary bone. These results suggest that endocrine disruptors directly act on osteoblastic bone formation.5.RT-PCR analysis of vitellogenin mRNA expression in avian liver.We tried to detect vitellogenin mRNA expression in liver of male quails treated with estrogen receptor agonists. As a result, vitellogenin mRNA expression was induced by estrogen receptor α agonist but not estrogen receptor β agonist. This indicates that vitellogenin synthesis in liver is stimulated through estrogen receptor α. Less

1.芳烃受体（AhR ;二恶英受体）mRNA表达与衰老的关系采用RT-PCR方法检测了0 ~ 390日龄雌性雏鸡不同组织中AhR mRNA的表达。结果表明，在各个阶段的砂囊、肝、肺、肾和心脏中均检测到AhR mRNA的表达。而母鸡输卵管和骨髓中AhR mRNA在0日龄和30-90日龄分别为零和少量表达。这些结果表明，蛋鸡输卵管和骨髓对二恶英的敏感性随日龄的变化而变化。2. AhR在蛋鸡输卵管中的表达与定位：Western blotting检测到AhR在鸡输卵管的顶腺、峡部和壳腺中均有表达。尤其是峡部和壳腺，AhR强烈定位于上皮细胞的细胞质中。这些结果表明，二恶英直接破坏了峡部和壳腺的代谢，导致蛋壳弱化 ...更多信息用DDE（10 mg/d）处理雄性日本鹌鹑7天，可诱导股骨骨髓腔形成髓骨。同时，治疗增加血清卵黄蛋白原水平。这些结果表明，鸟类髓质骨形成作为内分泌干扰物的体内生物测定模型具有上级优势。4.内分泌干扰物受体在鸟类髓质骨中的表达与定位雌激素受体（α和β）和雄激素受体是内分泌干扰物的受体。本研究通过原位杂交和免疫组织化学分析，发现两种受体（尤其是雌激素受体α亚型）均定位于鸡骨髓成骨细胞。这些结果表明，内分泌干扰物直接作用于成骨细胞的骨形成。5.禽肝脏卵黄蛋白原mRNA表达的RT-PCR分析结果表明，雌激素受体α激动剂能诱导卵黄蛋白原mRNA的表达，而雌激素受体β激动剂对卵黄蛋白原mRNA的表达无明显影响。这表明雌激素受体α刺激肝脏卵黄蛋白原的合成。少