Role of nonnuclear estrogen receptor GPR30 in preganglionic vagal neurons

非核雌激素受体 GPR30 在节前迷走神经元中的作用

基本信息

批准号：
8217285
负责人：
EUGEN BRAILOIU
金额：
$ 29.7万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-02-01 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8217285
关键词：
Address Agonist Aromatase Blood Pressure Blood Pressure Monitors Bradycardia Brain Stem Calcium Cardiac Cardiovascular Diseases Cardiovascular Physiology Cardiovascular system Cells Clinical Conjugated Equine Estrogens Development Estradiol Estrogen Nuclear Receptor Estrogen Receptors Estrogens Goals Health Heart Heart Rate Hormone replacement therapy Hypothalamic structure Image Immunohistochemistry In Vitro Knowledge Label Link Measurable Measurement Mediating Membrane Microinjections Morbidity - disease rate Neurons Nuclear Receptors Pathway interactions Postmenopause Premenopause Progesterone Property Rattus Regulation Relative (related person)Role Signal Transduction Slice Source Staining method Stains Synapses Techniques Testing Therapeutic Therapeutic Agents Urethane Vagus nerve structure Woman cholinergic neuron design extracellular immunocytochemistry in vivo insight interdisciplinary approach intravenous administration mortality neural circuit novel nucleus ambiguus patch clamp pressure protective effect release of sequestered calcium ion into cytoplasm research study response

项目摘要

DESCRIPTION (provided by applicant): Cardiovascular disease is a leading cause of morbidity and mortality. Estrogen has been proposed to have a protective effect. In addition to the interaction with two classical nuclear receptors, estrogen activates a non nuclear receptor, GPR30. Our long-term goal is to understand the role of GPR30 in estrogen-mediated neuronal effects. The objective of this application is to determine the role of GPR30 in cardiovascular regulation. Our central hypothesis is that estrogen acting on GPR30 located on neurons of the nucleus ambiguus increases vagal activity to the heart, which may be cardioprotective. Viewed in this context, GPR30 is a potential target for the development of novel cardiovascular therapeutic agents. We will use a multidisciplinary approach including immunohistochemistry, confocal imaging, in vivo monitoring of blood pressure and heart rate, fluorimetric measurement of cytosolic calcium and whole-cell patch-clamp recording to address the following aims. First, localization of GPR30 to cardiac preganglionic neurons of the nucleus ambiguus. Our preliminary results indicate that GPR30 is present in cholinergic neurons of the nucleus ambiguus. We will use retrograde labeling, single and double immunohistochemical staining to identify the presence of GPR30 in cardiac projecting neurons of the nucleus ambiguus and its colocalization with aromatase. Second, measurement of heart rate and blood pressure in response to GPR30 agonists in vivo. Our preliminary results indicate that microinjection of G-1, a specific GPR30 agonist, or 17-estradiol (E2) into the nucleus ambiguus produces bradycardia; the effect was abolished by prior administration of G-36, a GPR30 antagonist. In addition, pilot experiments indicate that intravenous administration of G-1 and E2 decreased mean arterial pressure and heart rate in urethane-anesthetized rats. We will determine the effect of intravenous administration of GPR30 agonists on blood pressure, heart rate and the participation of the vagus nerve in these responses. Third, electrophysiological response of GPR30 activation in single nucleus ambiguus neurons in vitro. Our preliminary results indicate that G-1 excites neurons from nucleus ambiguus. We will determine the effect of GPR30 agonists on membrane properties, evoked and spontaneous synaptic currents from nucleus ambiguus neurons using whole-cell patch-clamp recordings in brainstem slices. Fourth, define the calcium pathways involved in GPR30-mediated neuronal responses. Our preliminary results indicate that activation of GPR30 produces calcium mobilization from external and internal sources in cultured rat hypothalamic neurons; the effect was abolished by pretreatment with G-36. We will determine the calcium pools involved in GPR30 signaling in premotor cardiac vagal neurons. The result of this study will extend our understanding of the mechanisms of action of estrogen on neurons involved in cardiovascular regulation with implications for the development of effective therapeutic approaches in cardiovascular disorders. PUBLIC HEALTH RELEVANCE: We will study the role of a new estrogen receptor in the central control of cardiovascular function. A better understanding of the mechanisms of action of estrogen is critical for the development of new treatments for cardiovascular disease.

描述（由申请人提供）：心血管疾病是发病和死亡的主要原因。雌激素被认为具有保护作用。除了与两种经典核受体相互作用外，雌激素还激活一种非核受体 GPR30。我们的长期目标是了解 GPR30 在雌激素介导的神经元效应中的作用。本申请的目的是确定 GPR30 在心血管调节中的作用。我们的中心假设是，作用于疑核神经元上的 GPR30 的雌激素会增加心脏的迷走神经活动，这可能具有心脏保护作用。从这个角度来看，GPR30是开发新型心血管治疗药物的潜在靶点。我们将采用多学科方法，包括免疫组织化学、共聚焦成像、血压和心率体内监测、胞质钙荧光测量和全细胞膜片钳记录来实现以下目标。首先，GPR30 定位于心脏疑核的节前神经元。我们的初步结果表明 GPR30 存在于疑核的胆碱能神经元中。我们将使用逆行标记、单免疫组织化学染色和双免疫组织化学染色来鉴定疑核心脏投射神经元中 GPR30 的存在及其与芳香酶的共定位。其次，测量体内对 GPR30 激动剂反应的心率和血压。我们的初步结果表明，将 G-1（一种特定的 GPR30 激动剂）或 17-雌二醇 (E2) 显微注射到疑核中会产生心动过缓；预先给予 G-36（一种 GPR30 拮抗剂）可消除该效应。此外，初步实验表明，静脉注射 G-1 和 E2 可以降低乌拉坦麻醉大鼠的平均动脉压和心率。我们将确定静脉注射 GPR30 激动剂对血压、心率以及迷走神经参与这些反应的影响。第三，体外单核疑神经元GPR30激活的电生理反应。我们的初步结果表明 G-1 兴奋来自疑核的神经元。我们将使用脑干切片中的全细胞膜片钳记录来确定 GPR30 激动剂对膜特性、疑核神经元诱发和自发突触电流的影响。第四，定义参与 GPR30 介导的神经元反应的钙途径。我们的初步结果表明，GPR30 的激活会在培养的大鼠下丘脑神经元中产生来自外部和内部来源的钙动员； G-36预处理消除了该作用。我们将确定参与运动前心脏迷走神经元 GPR30 信号传导的钙池。这项研究的结果将加深我们对雌激素对心血管调节神经元作用机制的理解，对开发心血管疾病的有效治疗方法具有重要意义。公共健康相关性：我们将研究新雌激素受体在心血管功能中央控制中的作用。更好地了解雌激素的作用机制对于开发心血管疾病新疗法至关重要。

项目成果

期刊论文数量（11）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Direct evidence of intracrine angiotensin II signaling in neurons.

神经元内分泌血管紧张素 II 信号传导的直接证据。

DOI：
10.1152/ajpcell.00131.2013
发表时间：
2014-04-15
期刊：
American journal of physiology. Cell physiology
影响因子：
0
作者：
Elena Deliu;G. Brailoiu;S. Eguchi;N. Hoffman;J. Rabinowitz;D. Tilley;M. Madesh;W. Koch;E. Brǎiloiu
通讯作者：
E. Brǎiloiu

Convergent regulation of the lysosomal two-pore channel-2 by Mg²⁺, NAADP, PI(3,5)P₂ and multiple protein kinases.

Mg2α、NAADP、PI(3,5)Pα 和多种蛋白激酶对溶酶体双孔通道 2 的聚合调节。

DOI：
发表时间：
2014-03-03
期刊：
影响因子：
0
作者：
Jha, Archana;Ahuja, Malini;Patel, Sandip;Brailoiu, Eugen;Muallem, Shmuel
通讯作者：
Muallem, Shmuel

Urocortin 3 elevates cytosolic calcium in nucleus ambiguus neurons.

尿皮质素 3 会升高疑核神经元中的胞质钙。

DOI：
发表时间：
2012-09
期刊：
影响因子：
4.7
作者：
Brailoiu, G Cristina;Deliu, Elena;Tica, Andrei A;Chitravanshi, Vineet C;Brailoiu, Eugen
通讯作者：
Brailoiu, Eugen

An NAADP-gated two-pore channel targeted to the plasma membrane uncouples triggering from amplifying Ca2+ signals.

靶向质膜的 NAADP 门控双孔通道可解除放大 Ca2 信号的触发。

DOI：
发表时间：
2010-12-03
期刊：
影响因子：
0
作者：
Brailoiu, Eugen;Rahman, Taufiq;Churamani, Dev;Prole, David L;Brailoiu, G Cristina;Hooper, Robert;Taylor, Colin W;Patel, Sandip
通讯作者：
Patel, Sandip

Membrane topology of NAADP-sensitive two-pore channels and their regulation by N-linked glycosylation.

NAADP 敏感双孔通道的膜拓扑及其通过 N 连接糖基化的调节。