EXTRAHYPOTHALAMIC AVP: SYNTHESIS AND SECRETION

下丘脑外 AVP：合成和分泌

• 批准号: 6540236
• 负责人: ADAM CHODOBSKI
• 金额: $ 26.16万
• 依托单位: RHODE ISLAND HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-26 至 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6540236
• 关键词: arginine vasopressin; cerebrospinal fluid; choroid plexus; gene expression; glia; hormone biosynthesis; immunocytochemistry; laboratory rat; neuroendocrine system; neurohormones; neurons; neuropathology; polymerase chain reaction; protein kinase; radioimmunoassay; secretion; statistics /biometry; tissue /cell culture; western blottings

Choroid plexus (CP), a major generator of cerebrospinal fluid (CSF), produces and secretes into the CSF various neuropeptides, growth factors, and cytokines. Our general working hypothesis is that the CP-derived polypeptides not only regulate the function of CP epithelium locally, by autocrine actions, but also exert distal paracrine effects on target cells in the brain. The distal brain regions may be reached through CSF pathways due to continual production and flow of CSF. The focus of this application is on arginine vasopressin (AVP). CSF-borne AVP plays an important role in promoting cerebral edema and vasospasm that accompany several CNS disorders, including trauma, subarachnoid hemorrhage (SAH), and ischemia. However, the sources of CSF AVP and the regulation of AVP release into the CSF are not well understood. Our previous studies have demonstrated that CP epihelium is a likely source of CSF AVP. Specific aims of the present proposal are to obtain an insight into 1) the regulation of choroidal AVP gene expression, 2) intracellular mediators and neurohormonal secretagogues promoting AVP release from CP epithelium, and 3) the nature of this secretory process (via either the regulated or constitutive pathway). We will approach the above questions by applying both in vivo rat model and in vitro CP epithelial cell cultures. The functional experiments on AVP release will be complemented with molecular and immuno-chemical analyses, such as reverse transcriptase- polymerase chain reaction, Western blotting, and immuno- cytochemistry. Radioimmunoassay will be used to quantify AVP content. The long-term objective is to obtain a comprehensive view of how the CP-CSF system regulates the internal milieu for neuronal and glial cells. These studies will form the basis for development of pathological models and, consequently, effective therapies for the treatment of CNS disorders, such as trauma, SAH, and stroke.

脉络丛 (CP) 是脑脊液 (CSF) 的主要产生者，产生并向 CSF 分泌各种神经肽、生长因子和细胞因子。 我们的一般工作假设是，CP衍生的多肽不仅通过自分泌作用局部调节CP上皮的功能，而且还对大脑中的靶细胞发挥远端旁分泌作用。 由于脑脊液的持续产生和流动，可以通过脑脊液通路到达远端大脑区域。该应用的重点是精氨酸加压素 (AVP)。脑脊液携带的 AVP 在促进脑水肿和血管痉挛方面发挥着重要作用，这些脑水肿和血管痉挛伴随着多种中枢神经系统疾病，包括创伤、蛛网膜下腔出血 (SAH) 和缺血。 然而，CSF AVP 的来源以及 AVP 释放到 CSF 中的调节尚不清楚。 我们之前的研究表明，CP 上皮细胞可能是 CSF AVP 的来源。本提案的具体目的是深入了解 1) 脉络膜 AVP 基因表达的调节，2) 促进 CP 上皮释放 AVP 的细胞内介质和神经激素促分泌素，以及 3) 这种分泌过程的性质（通过调节途径或组成途径）。我们将通过应用体内大鼠模型和体外 CP 上皮细胞培养来解决上述问题。 AVP 释放的功能实验将辅以分子和免疫化学分析，例如逆转录酶-聚合酶链反应、蛋白质印迹和免疫细胞化学。 放射免疫测定法将用于量化 AVP 含量。长期目标是全面了解 CP-CSF 系统如何调节神经元和神经胶质细胞的内部环境。 这些研究将为病理模型的开发奠定基础，从而为治疗中枢神经系统疾病（如创伤、蛛网膜下腔出血和中风）的有效疗法奠定基础。