DETERMINATION OF DNA SEQUENCE OF NEUROPEPTIDE Y Y3-RECEPTOR

为了探讨神经源性肺水肿的发病机制，我们利用了三个实验：(1)神经肽Y Y3受体的克隆及其DNA序列测定；(2)血管内皮细胞的参与；(3)中枢神经系统的作用。我们已报道神经肽Y是交感神经末梢释放的一种神经递质，它可能增加纤维蛋白所致肺水肿发展过程中的肺血管通透性。因此，它扰乱了一代新药。我们提取了MR*，并构建了cDNA。将该基因与ZAP Express载体连接，构建了cDNA文库。用含有该基因的噬菌体感染E.ColiDNA。培养后，大肠杆菌在琼脂平板上成功地形成了空斑。用ANT*NPY抗体和抗PYY抗体处理印迹的硝酸纤维素膜。通过彩色化可视化，c…更多的隆宁正在研究中。第二，我们利用大鼠主动脉内皮细胞，使细胞在琼脂平板上存活。血管内皮细胞现在被研究用荧光分子结合白蛋白等渗透性分子来检测血管的通透性。同时，我们还研究了血管内皮细胞生长因子(VEGF)的发展，使血管床对大分子白蛋白具有通透性。静脉麻醉剂、氯胺酮、异丙酚和硫喷妥钠、神经递质、神经肽Y、去甲肾上腺素和*乙酰胆碱以及其他生物活性物质。氯胺酮和硫喷妥钠具有增加血管内皮细胞释放血管内皮生长因子的活性。我们已经报道，延髓中释放的一氧化氮促进了神经源性肺水肿的发展。*由于左侧迷走神经切断术在2周后增加了NO合成酶，我们对纤维蛋白诱导的肺水肿进行了检测，获得了*结果表明NO对水肿的发展有抑制作用。然而，血管通透性的增加在2周时达到最大，但在4周时更明显。现在，我们正在研究1-谷氨酸，一种可兴奋的神经递质，在纤维蛋白诱导的肺水肿中的作用。较少

In order to investigate a mechanism involved in the development of neurogenic pulmonary edema, we utilized three experiments: (1) cloning of neuropeptide Y Y3-receptor and determination of its DNA sequenc* (2) involvement of vascular endothelial cells; (3) action of central nervous system.We have reported that neuropeptide Y is a neurotransmitter, released from the sympathetic nerve terminal, which might increa* the pulmonary vascular permeability in the development of fibrin-induced pulmonary edema, a model of neurogenic pulmonary edema* So far, DNA sequence of NPY Y3-receptor has not been elucidated. Therefore it disturbs a generation of new drug. We extracted mR* and created cDNA. The cDNA was ligated to ZAP Express Vector, creating cDNA library. E.coli DNA was infected by phage which h* the cDNA. After incubation, E. coli successfully made plaques on agar plate. Blotted nitrocellulose membrane was treated with ant* NPY antibody and anti PYY antibody. Visualized with colorization, c … More loning is now under study.Secondly, we utilized rat aortic endothelial cells, making the cells alive in agar plates. Endothelial cells are now investigated f* detecting the vascular permeability with permeating molecules such as fluorescent molecule-binding albumin. At the same time, we * also studying the development of vascular endothelial cell growth factor (VEGF) which makes the vascular bed permeable to large * molecule, albumin. Venous anesthetics, ketamine, propofol, and thiopental, neurotransmitters, neuropeptide Y, norepinephrine and * acetylcholine, and other bioactive substances. Ketamine and thiopental have an activity to increase the release ofVEGF from the * vascular endothelial cells.We already reported that NO release in the medulla oblongata has enhanced the development of neurogenic pulmonary edema. * Since unilateral left-vagotomy increase the NO synthase 2 weeks later, we have examined the fibrin-induced pulmonary edema, obtain * the results demonstrating the inhibitory action of NO on the edema development. However, the increase in vascular permeability was * maximum at 2 weeks time point, but rather increased more at 4 weeks. Now we are studying the effects of 1-glutamate, an excitable * neurotransmitter, on the fibrin-induced pulmonary edema. Less