Characterization of Central ABP/SHBG Receptors

中枢 ABP/SHBG 受体的表征

• 批准号: 6822232
• 负责人: JACK D CALDWELL
• 金额: $ 15.5万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6822232
• 关键词: affinity chromatography; androgen binding protein; binding sites; estradiol; fluorescence microscopy; high performance liquid chromatography; hormone binding protein; hypothalamus; laboratory rat; mass spectrometry; mitogen activated protein kinase; oxytocin; phosphorylation; polymerase chain reaction; progesterone; protein binding; protein protein interaction; steroid hormone; steroid hormone receptor; voltage /patch clamp

DESCRIPTION (provided by applicant): An emerging model of action for steroid-binding proteins is that they are essential in delivering steroids to either steroid membrane or cytoplasmic receptors. Whereas the more commonly known model suggests that steroid-binding proteins such as sex hormone binding globulin (SHBG) are only made in the periphery and only deliver steroids to be passively released at the target cell, we have found that SHBG is produced in the brain where it facilitates female sexual receptivity. Therefore, we have two ends of the answer to the question of whether SHBG is a neuromodulator: its production and its behavioral effects. However, we need to find out whether and where there are receptors for SHBG in brain. In generating preliminary data, we have found that SHBG stimulates female sexual receptivity and that coupling it to dihydrotestosterone blocks this facilitative effect. We have found that microiontophoresing SHBG onto hypothalamic tissue sections results in an immediate depolarization of magnocellular neurons. We have found that cells transfected with either estradiol receptor alpha or beta respond to both SHBG and SHBG-estradiol application with an elevation of MAP kinase phosphorylation. Using SELDI-TOF mass spectrometry on affinity chromatography eluted hypothalamic tissue, we have identified protein peaks at 144000 and 168000 daltons that seem to represent SHBG receptors. We have found SHBG produced in hypothalamic areas in the same neurons as the reproductively important peptide oxytocin. Both SHBG and oxytocin are found in varicosities and in synaptic vesicles suggesting they are released within the brain. Therefore, work done in several laboratories, collaborating together via the PI, has found that SHBG is made in brain, may be released in brain, [and] has important very rapid actions on neurons and cells in vitro, and so we would like funding to search for receptors for this potentially very important brain protein. This application proposes a confluence of particular scientific expertise including the PI and Drs. Gustav Jirikowski at the University of Jena, Germany, Jeffrey Tasker of Tulane University and Robert Shapiro at the Oregon Health Sciences University. In the words of one of the reviewers of the initial application, "The mechanism mediating the delivery of steroid hormones to their receptors has important implications for mental health, as well the action of steroid hormones in cancer, heart disease, and other disorders." This proposal examines this mechanism.

描述（由申请人提供）：类固醇结合蛋白的新兴作用模型是，它们对于将类固醇传递到类固醇膜或细胞质受体至关重要。虽然更为众所周知的模型表明，类固醇结合蛋白（例如性激素结合球蛋白（SHBG））仅在外围制造，并且仅提供在靶细胞处被动释放的类固醇，但我们发现SHBG是在促进女性性爱的大脑中产生的。因此，我们对SHBG是否是神经调节剂的问题有两个目的：它的产生及其行为影响。但是，我们需要找出大脑中SHBG的受体是否以及位置。在生成初步数据时，我们发现SHBG刺激了女性的性接受能力，并且将其与二氢睾丸激素耦合会阻止这种促进作用。我们发现，将SHBG的微滴体上的微滴体上的下丘脑组织切片导致巨细胞神经元立即去极化。我们发现，用雌二醇受体α或β转染的细胞对SHBG和SHBG-雌二醇的应用反应，并升高MAP激酶磷酸化。使用Seldi-TOF质谱法上的亲和色谱洗脱了下丘脑组织，我们已经确定了似乎代表SHBG受体的144000和168000 daltons的蛋白质峰。我们发现与生殖重要的肽催产素在同一神经元的下丘脑区域产生的SHBG。 SHBG和催产素都在静脉曲张和突触囊泡中都发现，表明它们在大脑内释放。因此，通过PI合作的几个实验室完成的工作发现SHBG是在大脑中释放的，可以在大脑中释放，并且在体外对神经元和细胞具有重要的快速作用，因此我们希望资金以寻找这种潜在非常重要的脑蛋白的受体。该应用提出了包括PI和DRS在内的特定科学专业知识的汇合。德国耶拿大学的古斯塔夫·吉里科夫斯基（Gustav Jirikowski），杜兰大学（Tulane University）的杰弗里·塔斯特（Jeffrey Tasker）和俄勒冈健康科学大学的罗伯特·夏皮罗（Robert Shapiro）。用最初应用的一位审稿人的话说：“介导类固醇激素向其受体传递的机制对心理健康具有重要意义，以及类固醇激素在癌症，心脏病和其他疾病中的作用。”该提案检查了这种机制。