LOCATION OF PORE SYSTEMS IN CAPILLARY WALLS

描述(改编自《调查者摘要》)：血管 血管内皮细胞在哺乳动物和人类的生理中起着关键作用 包括在内。它调节大量的、持续的水和亲水性的交换 和疏水溶质(小分子和大分子)之间 血浆和间质液体。所有细胞的存活率 身体的组织和器官依赖于这些持续的交换。它 参与控制凝血和抗凝过程 影响血液。它在早期阶段起着至关重要的作用。 炎症反应。它参与了正常和病理的 新生血管(血管生成)，包括肿瘤血管生成 取决于肿瘤的生长和转移。在这些方面，内皮细胞 故障是大量人类的开始基线 病理情况。根据这笔赠款所做的工作已经提供了 关于细胞和分子相互作用的有用信息 上面列出的关键功能。它还开辟了新的前景， 首席调查员建议在继续调查期间 奖助金。他相信，增加对这些功能的理解将 帮助预防、缓解并在可能的情况下治愈某些内皮功能障碍 未来的时间。这些研究是在五个具体目标下提出的。我。 在第一个特定目标中，使用α-酸性糖蛋白的实验 (类红粘蛋白)作为示踪剂被提出，以加强 连续性内皮细胞大分子处理中的细胞穿透作用 以细胞间连接为界。形态和细胞分级 将采用多种方法来研究示踪剂通过 PVS或小窝融合形成的跨内皮细胞通道。尝试 将被用来识别两个洞穴种群，即小的和大的 一种，容纳40埃和250埃的二硝基苯基探针， 分别进行了分析。这些研究还将扩大，以确定 血浆蛋白(S)，锚定(S)α-酸性糖蛋白和 构成纤维基质。进一步的实验以确定因素(S) 文中还提出了小窝融合的原因。在这些 实验中，融合蛋白将利用含有以下成分的载体产生 NEM敏感因子(NSF)和SNAPs的cDNA；它们对应的 将产生多克隆抗体，并用于研究其分布 在血管内皮细胞隔间。分配情况将通过以下方式确认 使用其他方法，例如使用特定的单抗 抗体。二.在第二个具体目标中，拟议的研究将 重点关注带或不带横隔膜的有窗内皮细胞，并利用 具体目标#一、三、第三个具体目标所述的方法 涉及黄曲霉毒素诱导的小窝结构调制的研究 血小板活化因子与血管内皮生长因子 (血管内皮生长因子)。我们会特别注意研究 下腔静脉旁糖蛋白组合，由PI定义为“紧身衣”。 PAF和血管内皮生长因子向血管系统的灌流。因为他们的行为是 通过它们的受体，这些实验被概述为研究 这些生长因子影响下的受体磷酸化。这个 研究将扩展到调查负面的和 可能的正向跨细胞调节因子，包括NEM， 钠-硝普钠、心钠素和钙离子载体。在这些实验中， 如果有内皮间连接的改变，将进行调查。 四.具体目标#4涉及生物化学特性 肺组织中富含空泡部分的糖蛋白 内皮，利用阳离子二氧化硅程序。他们的孤立将会是 通过使用各种单抗促进了这一过程。这些研究 将被扩展以描述功能的多样性 小窝。五.在具体目标#5中，窗口化的特征 来自横隔膜毛细血管的内皮，例如管周毛细血管 将通过利用上述特定的技术来实现 目标。

DESCRIPTION (Adapted from Investigator's Abstract): The vascular endothelium performs critical functions in the physiology of mammals, men included. It mediates vast, continuous exchanges of water and hydrophilic and hydrophobic solutes (small as well as large molecules) between the blood-plasma and interstitial fluids. The survival of all cells of the tissues and organs of the body depends on these continuous exchanges. It participates in the control of coagulant and anticoagulant processes affecting the blood. It plays an essential role in the initial phases of inflammatory reactions. And it is involved in normal and pathological neovascularization (angiogenesis), including tumor vascularization on which depends tumor growth and metastasis. On these accounts, endothelial malfunction is the starting baseline for a large number of human pathological conditions. Work done under this grant has already provided useful information on cellular and molecular interactions involved in each of the critical functions listed above. It has also opened new vistas which the Principal Investigator proposes to explore during the continuation of the grant. He believes that increased understanding of these functions will help prevent, mitigate and if possible cure endothelial malfunctions at some time in the future. The studies are proposed under five Specific Aims. I. In the first specific aim, experiments utilizing alpha1-acidic glycoprotein (orosomucoid) as a tracer are proposed to reinforce the concept/process of transcytosis in the handling of macromolecules in continuous endothelia bounded by intercellular junctions. Morphological and cell fractionation methods will be employed to study the entrance and exit of the tracer via transendothelial channels formed by the fusion of Pvs or caveolae. Attempts will be made to identify two populations of caveolae, i.e., small and large ones, accommodating 40 angstrom and 250 angstrom dinitrophenylated probes, respectively. The studies will be extended also to identify the plasmalemmal protein(s) which anchor(s) the alpha1-acidic glycoproteins and constitute the fiber matrix. Further experiments to identify the factor(s) responsible for the fusion of the caveolae are also proposed. In these experiments, fusion proteins would be generated utilizing vectors containing cDNA for NEM sensitive factor (NSF) and SNAPs; their corresponding polyclonal antibodies will be generated, and used to study the distribution in the endothelial compartments. The distribution would be confirmed by using other approaches, such as, with the employment of specific monoclonal antibodies. II. In the second specific aim, the studies proposed will focus on fenestrated endothelia with or without diaphragms, and utilize the methodologies as described in specific aim # I. III. The third specific aim relates to the studies of structural modulations of caveolae induced by platelet activating factor (PAF) and vascular endothelial growth factor (VEGF). A particular attention will be paid to study the changes in the paracaveolar glycoprotein assembly, defined by the PI as "corset", upon perfusion of PAF and VEGF into the vasculature. Since their actions are mediated via their receptors, the experiments are outlined to study the receptor phosphorylation under the influence of these growth factors. The studies will be extended to investigate the influence of negative and possible positive modulators of transcytosis, and they include NEM, Na-nitroprusside, ANP and Ca++ ionophores. In these experiments, alterations, if any, in the interendothelial junctions will be investigated. IV. The specific aim # 4 deals with the biochemical characterization of glycoproteins enriched in the caveolar fractions isolated from pulmonary endothelia, utilizing cationized silica procedures. Their isolation will be facilitated by the use of various monoclonal antibodies. These studies would be extended to characterize the multiplicity of the functions of caveolae. V. In specific aim # 5, the characterization of fenestrated endothelia from diaphragmatic capillaries, e.g., peritubular capillaries will be carried out by utilizing the techniques described in above specific aims.