Mechanisms Regulating Gastrointestinal Hormone Secretion

调节胃肠激素分泌的机制

• 批准号: 8258993
• 负责人: Rodger A. Liddle
• 金额: $ 34.15万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-17 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8258993
• 关键词: Address; Amino Acids; Animals; Apical; Aromatic Amino Acids; Attention; Bile fluid; Blood; Brain; Calcium; Calcium Channel; Calcium Signaling; Calcium-Sensing Receptors; Cell physiology; Cells; Cholecystokinin; Cholecystokinin Receptor; Coupled; Data; Digestion; Distant; Eating; Endocrine; Enzymes; Fatty Acids; Fluorescence; Fluorescence Microscopy; Fluorescence-Activated Cell Sorting; Food; Gallbladder; Gastrointestinal Hormones; Gastroparesis; Green Fluorescent Proteins; Hormonal; Hormones; Human; In Vitro; Individual; Ingestion; Intestinal Mucosa; Intestines; Ion Channel; Lead; Measurement; Measures; Mediating; Membrane Potentials; Methods; Mus; Neuroendocrine Cell; Nutrient; Pancreas; Pancreatic enzyme; Phenylalanine; Physiological; Potassium Channel; Process; Property; Receptor Activation; Regulation; Role; Satiation; Second Messenger Systems; Signal Pathway; Small Intestines; Stomach; Stream; Study models; Surface; Techniques; Transgenic Mice; Tryptophan; Vagus nerve structure; enhanced green fluorescent protein; extracellular; gastrointestinal; hormone regulation; in vivo; increased appetite; insight; intestinal epithelium; mouse model; new technology; novel; patch clamp; prevent; receptor; second messenger; tool

DESCRIPTION (provided by applicant): Gastrointestinal hormones are produced by discrete neuroendocrine cells which are scattered throughout the intestine. Most GI hormone-containing cells reside within the intestinal mucosa and are often oriented with their apical region open to the lumen of the intestine. Cholecystokinin (CCK) is a prototypical gastrointestinal hormone that regulates gallbladder contraction, pancreatic enzyme secretion, delays gastric emptying, and induces satiety. As is typical of most GI hormones, CCK is secreted into the blood stream after ingestion of a meal. It is generally believed that nutrients stimulate CCK release but the cellular mechanisms regulating CCK cell function are largely unknown. Recently the PI has developed a method for isolating and characterizing individual, viable, native intestinal CCK cells and by highly enriching these cells it has been possible to study CCK secretion in vitro, identify receptors on these cells and investigate second messenger signaling pathways involved in regulated hormone secretion. Together these approaches have the ability to provide unique insights into the mechanisms by which nutrients may stimulate CCK secretion. Importantly, the PI has preliminary data that CCK cells express the calcium-sensing receptor (CaSR) and that CaSR mediates amino acid-induced CCK secretion. The PI will use complementary techniques to study the regulation of hormone secretion. These include: (1) isolation and identification of native CCK cells, (2) measurements of CCK secretion in vivo and in vitro, (3) quantification of intracellular calcium fluorescence, and (4) characterization of electrophysiological properties measured by whole-cell patch clamp recordings. The central hypothesis of this application is that gastrointestinal hormone secreting cels are electrically excitable cells whose secretion is regulated by receptor and ion channel activation. The overall purpose of this proposal is to understand the physiological regulators of GI endocrine cells with the initial focus on how amino acids control CCK secretion. Characterization of CaSR and its relationship to ion channel activation on CCK cells will be addressed by the following Specific Aims: 1. To characterize the role of CaSR in the regulation of CCK secretion in isolated CCK cells in vitro and in mice in vivo. 2. To determine effects of CaSR activation on calcium signaling in CCK cells. 3. To characterize the electrophysiological properties of CCK cells and evaluate CaSR regulation of membrane potential, and potassium channel and calcium channel activities. Each of these aims will focus on regulation of CaSR as a critical step in the regulation of amino acid-stimulated CCK secretion. More globally, these aims should provide considerable insight into the mechanisms by which GI endocrine cells are regulated by nutrients known to be important in the control of hormone secretion. PUBLIC HEALTH RELEVANCE: Cholecystokinin (CCK) is a major gastrointestinal hormone that is important in many digestive processes. Upon ingestion of a meal, CCK is released from the upper small intestine and acts on the pancreas to secrete digestive enzymes, the gallbladder to release bile, the stomach to prevent excess delivery of food to the intestine, and the brain through the vagus nerve to reduce food intake. Together these actions coordinate the ingestion and digestion of food. Despite these important effects, little is known about the control of CCK secretion. Specifically, the cellular mechanisms by which nutrients regulate CCK secretion are largely unknown. With new technologies that are now available to the PI, the current proposal will define and characterize a receptor that explains how amino acids stimulate hormone secretion. This receptor is coupled to regulation of ion channels that are involved in regulating CCK secretion. These studies should greatly expand our understanding of the nutrient regulation of hormone secretion. It is conceivable that these studies could lead to novel (luminally directed) therapies to regulate CCK release and, thereby, distant effects of CCK throughout the body (e.g., gallbladder contraction, satiety, etc.).

性状（由申请人提供）：胃肠激素由分散在整个肠道中的离散神经内分泌细胞产生。大多数含GI酶的细胞位于肠粘膜内，并且通常以其顶端区域向肠腔开放的方式定向。胆囊收缩素（Cholecystokinin，CCK）是一种典型的胃肠道激素，可调节胆囊收缩、胰腺酶分泌、延迟胃排空和诱导饱腹感。作为大多数胃肠道激素的典型，CCK在进食后分泌到血液中。一般认为，营养素刺激CCK释放，但调节CCK细胞功能的细胞机制在很大程度上是未知的。最近，PI开发了一种分离和表征单个、活的、天然肠CCK细胞的方法，通过高度富集这些细胞，可以在体外研究CCK分泌，鉴定这些细胞上的受体，并研究参与调节激素分泌的第二信使信号通路。总之，这些方法有能力提供独特的见解营养素可能刺激CCK分泌的机制。重要的是，PI有初步数据表明CCK细胞表达钙敏感受体（CaSR），CaSR介导氨基酸诱导的CCK分泌。PI将使用补充技术研究激素分泌的调节。其中包括：（1）天然CCK细胞的分离和鉴定，（2）体内和体外CCK分泌的测量，（3）细胞内钙荧光的定量，和（4）通过全细胞膜片钳记录测量的电生理特性的表征。本申请的中心假设是胃肠激素分泌细胞是电可兴奋的细胞，其分泌受受体和离子通道激活的调节。本提案的总体目的是了解GI内分泌细胞的生理调节剂，最初的重点是氨基酸如何控制CCK分泌。CaSR的表征及其与CCK细胞上离子通道活化的关系将通过以下具体目的来解决：1.研究钙调素受体（CaSR）在离体CCK细胞和小鼠体内CCK分泌调节中的作用。 2.探讨CaSR激活对CCK细胞钙信号转导的影响。 3.研究CCK细胞的电生理特性，评价CaSR对膜电位、钾通道和钙通道活性的调节作用。这些目标中的每一个都将集中在CaSR的调节上，作为调节氨基酸刺激的CCK分泌的关键步骤。更广泛地说，这些目标应该提供相当多的洞察GI内分泌细胞的调节机制，营养素已知是重要的激素分泌的控制。 公共卫生相关性：胆囊收缩素（CCK）是一种主要的胃肠道激素，在许多消化过程中很重要。进食后，胆囊收缩素从小肠上部释放，作用于胰腺以分泌消化酶，作用于胆囊以释放胆汁，作用于胃以防止过量食物输送到肠道，并通过迷走神经作用于大脑以减少食物摄入。这些动作共同协调食物的摄取和消化。尽管有这些重要的作用，但对CCK分泌的控制知之甚少。具体而言，营养物质调节CCK分泌的细胞机制在很大程度上是未知的。利用PI现在可用的新技术，目前的提案将定义和表征解释氨基酸如何刺激激素分泌的受体。该受体与参与调节CCK分泌的离子通道的调节偶联。这些研究将极大地扩展我们对激素分泌的营养调节的理解。可以想象，这些研究可能导致新的（管腔定向）疗法来调节CCK释放，从而调节CCK在全身的远端效应（例如，胆囊收缩、饱腹感等）。