NYP INTERACTION WITH MICROVASCULR ENDOTHELIUM

神经肽Y是1982年发现的一种由36个氨基酸组成的多肽。 脑组织的提取液。自鉴定以来，有证据表明 一直在积累，以支持其在监管中的作用 心血管系统。这一活动的大部分被归因于 直接血管收缩及其突触后能力 增强其他血管收缩药的作用。其他报道的监管机构 功能涉及控制脑干区的NPY调节 自主神经流出到心脏和血管成分及其能力 调节去甲肾上腺素和其他递质的释放 血管周围神经、交感神经和内脏神经。 心脏水平。虽然有限，但有证据支持NPY 血管内皮细胞水平的调节。在大血管内皮细胞 模特们。已经观察到NPY在一只手中可以增强白细胞 以时间和浓度依赖的方式以及以 另一方面改造前列环素的生产。使用体外培养技术 牛源性毛细血管内皮细胞培养模型的建立 肾上腺髓质我们报告了高亲和力的存在 NPY及其类似物的特异性结合位点，并观察到 神经肽能够诱导相关的信号事件。 在其他模型系统中激活NPY受体。此外，我们 有初步数据支持可比的信令响应 来自牛视网膜的微血管组织，但不含内皮 来源于牛主动脉。我们目前的目标如下 建议有助于定义NPY互动的性质 微血管内皮细胞。我们的一般假设是 NPY受体优先分布于毛细血管内皮细胞 它们的激活导致了 内皮源性血管扩张剂。为了证明这一假说 我们建议落实以下具体目标：(一) 评估NPY受体是否在其他组织中优先表达 微血管。这将通过执行放射性配位体来实现 大鼠内皮细胞与培养内皮细胞的结合研究 血管和几种组织的微血管。(2)测试 NPY诱导心肌细胞释放NO和PGI2的假说 通过检测微血管内皮细胞的基础和 在存在和不存在的情况下刺激释放这些血管扩张剂 神经肽。(3)评价NPY诱导的相互作用 钙反应与其他信号级联反应暗示在 内皮功能的调节，特别是(A)参与 蛋白酪氨酸激酶通路的研究 蛋白激酶抑制剂的作用及其免疫检测 磷酸化蛋白对NPY受体激活的影响。(B)法律责任 钙激活K+通道的膜片钳效应 神经肽Y对电生理影响的实验研究 与这些通道的表达相关联的参数。(4) 检测NPY负偶联对腺苷环化酶是否有调节作用 测定血浆中NO和PGI2的释放量 这些特工在操纵后改变了cAMP水平。加起来 这些研究应有助于深入了解NPY的功能作用，网址为 微血管内皮细胞的水平扩大了我们的认识 对心血管系统的调节作用 神经肽。

NPY is a 36 amino acid peptide originally discovered in 1982 romextracts of brain tissue. Since its identification, evidence has been accumulating to favor its role in the regulationof the cardiovascular system. Most of this activity has been ascribed to both direct vasoconstriction and its ability to post-synaptically potentiate other vasoconstrictors action. Other reported regulatory functions concern NPY modulationof brainstem areas controlling the autonomic outflow to cardiac and vacular components and its ability to moduate the release of norepinephrine and other transmitters from perivascular nerves and the sympathetic and intrinsic nerves at the heart level. Although limited, there is evidence to support a NPY modulation at the endothelial level. In large vessel endothelial models. NPY has been observed in one hand to potentiate leukocyte adhesion in a time and concentration dependent manner and in the other hand to modify prostacyclin production. Using an in vitro model of cultured capillary endothelial cells derived from bovine adrenal medulla we have reported the presence of high affinity specific binding sites for NPY and its analogs and have observed that the neuropeptide is capable of inducing signaling events associated with NPY receptor activation in other model systems. Moreover, we have preliminary data to support a comparable signaling responses in microvascular tissue from bovine retina but not with endothelium derived from bovine aorta. Our present goal with the following proposal is to contribute to define the nature of NPY interaction with the microvascular endothelium. Our general hypothesis is that NPY receptors are preferentially distributed in capillary endothelium and that their activation results inthe formation of endothelialderived vasodilators. In order to prove this hypothesis we are propsing to carry out the following specific aims: (1) Evaluate whether NPY receptors are preferentially expressed in other microvasculatures. This will be achieved by performing radioligand binding studies with cultured endothelial cells dervied from large vessels and the microvasculature of several tissues. (2) Test the hypothesis that NPY induces the release of NO and PGI2 from microvascular endothelium through the detection of the basal and stimulated release of these vasodilators in the presence and absence of the neuropeptide. (3) Evaluate the interaction of NPY induced calcium response with other signaling cascades implied in the modulation of endothelial function, specifically (a) the involvement of protein tyrosine kinase pathways through the assessment of the effect of proteinkinases inhibitors and the immunodetectionof phosphorylated proteins upon NPY receptor activation. (b) therole of Ca2+-activated K+ channels through the performance of patch-clamp experiments to detect the effect of NPY on the electrophysiologic parameters associated with the expression of these channels. (4) Test whether the NPY negative coupling to adenylate cyclase modulates the stimulated release of NO and PGI2 by measuring the relese of these agents after manipulations to alter cAMP levels. Altogether the studies should provide insight into the functional role of NPY at the level of the microvscular endothelium expanding our understanding of the regulation of the cardiovascular system provided by this neuropeptide.