绵羊甲状腺自主合成并分泌褪黑素的研究  

Investigation into Synthesis and Secretion of Melatonin in Thyroid Gland in Sheep

作　　者：欧科鹏 李尤简 郭若婷 王颖[1] 顾真真[1] 刘乙[1] 刘小军[2] 

机构地区：[1]石河子大学动物科技学院,石河子832003 [2]河南农业大学牧医工程学院,郑州450002

出　　处：《畜牧兽医学报》2015年第4期576-582,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：教育部高等学校博士学科点专项科研基金(博导类)(20126518110004)

摘　　要：本研究旨在探讨绵羊甲状腺是否表达催化褪黑素合成的关键酶基因AANAT、HIOMT以及褪黑素受体基因MT1,并自主合成和分泌褪黑素,为研究甲状腺在绵羊季节性繁殖中的功能奠定基础。利用RT-PCR、组织免疫荧光、细胞免疫荧光和ELISA技术,分别在基因、蛋白质和细胞水平对绵羊甲状腺中合成褪黑素的关键酶基因AANAT、HIOMT以及褪黑素受体基因MT1的表达、细胞定位以及褪黑素的合成与分泌进行了研究。结果表明:绵羊甲状腺表达褪黑素合成酶基因AANAT、HIOMT和褪黑素受体基因MT1,并能够自主合成和分泌褪黑素。该研究首次证实了绵羊甲状腺具有自主合成并分泌褪黑素的功能,表明甲状腺内部可能存在有通过褪黑素调控甲状腺素合成的新通路,为进一步研究绵羊甲状腺自主合成褪黑素的功能奠定了基础。The objectives of present study were to investigate whether two key genes of enzymes for melatonin biosynthesis,arylalkylamine N-acetyltransferase(AANAT)and hydroxyindole-Omethyltransferase(HIOMT),and melatonin receptor MT1 were expressed in thyroid gland in sheep,and whether melatonin could be independently synthesized and secreted in the gland.The expression of AANAT,HIOMTand MT1 was firstly detected by using RT-PCR,and was then confirmed by immunofluorescenct histochemical assay in tissues of thyroid gland of sheep.The specificity of cultured primary thyroid gland cells was characterised by immunofluorescenct cytochemical assay.The synthesis and secretion of melatonin in sheep thyroid gland cells were analysed using ELISA.The results showed that AANAT,HIOMTand MT1 mRNAs were expressed in sheep thyroid tissue.Immunofluorescence assays confirmed that HIOMT and MT1 protein were localized on follicular cells in thyroid gland.Meanwhile,ELISA assay demonstrated that the thyroid gland of sheep could synthesize and secret melatonin autonomously.The present study evidenced,for the first time,that the melatonin synthesizing enzymes and melatonin receptor were presented in the thyroid tissue,and thyroid gland was able to synthesize and secret melatonin autonomously in sheep.The data provided new evidence for a putative novel intrathyroidal regulatory pathway that melatonin might be involved in thyroid-hormone synthesis.

关 键 词：甲状腺 褪黑激素 AANAT HIOMT MT1 绵羊 

分 类 号：S826[农业科学—畜牧学]