MECHANISMS INVOLVED IN LYMPHATIC CONTRACTILE ACTIVITY

我的长期职业目标是提高对以下问题的理解水平： 一个重要但知之甚少的器官的作用和机制 系统，淋巴系统。 我的近期职业目标是： 建立独立，高质量的科学研究的声誉; 获得终身职位;培养研究生和博士后研究员;以及 促进科学社会的进步，研究科 和编辑委员会。 获得这个奖项将有助于推进我的研究 计划通过提供激励措施，以学院腾出时间和 让私家侦探专注于科学研究 淋巴系统是 血液循环的一个专门部分，与血液循环密切相关。 维持正常的体液平衡。它被认为是 防止严重水肿形成的主要安全因素。 淋巴 系统还涉及营养吸收、淋巴细胞循环和 抗原呈递 这些任务是通过淋巴循环完成的。 液体通过淋巴系统主动泵送， 产生淋巴收缩。 这些的总体目标 研究将阐明参与生成的机制， 调节自发淋巴收缩。 以下具体 目的将被解决：1）确定内皮细胞的作用，平滑 肌肉细胞及其相互作用在压力敏感性的 淋巴泵; 2）确定内皮、平滑肌的作用 细胞及其在淋巴管流动敏感性中的相互作用 3）评估在体外循环期间淋巴细胞内钙的调节。 自发性收缩; 4）评估淋巴管的变化 在压力和流量引起的细胞内钙变化过程中， 淋巴泵; 5）评估淋巴起搏点活动的部位。 这些研究将在隔离、加压的收集中进行。 来自大鼠小肠肠系膜的消化物。 淋巴管 将从组织中分离出来并插管。 压力和流量 将通过改变入口来仔细地控制通过容器， 出口压力。 血管内皮和血管内皮-平滑肌的作用 肌肉的相互作用将使用技术进行研究，以抑制 和/或去除淋巴内皮。 淋巴收缩会 使用视频显微镜技术监测。 淋巴管直径将 在整个实验过程中持续测量。 从淋巴管 直径描记，淋巴泵的功能指数将被评估。 自发性高血压大鼠心肌细胞内钙离子水平的变化 将使用荧光钙敏感染料监测收缩 Fura-2 参与调节的细胞机制 将研究细胞内钙。 压力的影响， 还将评估淋巴钙调节的流量。 评价 淋巴管细胞内的膜电位将被 使用电位荧光染料Di-8-ANEPPS进行。 淋巴内起搏器活动的研究将 使用荧光成像技术进行。 的结果予以 调查应提供相当深入的机制， 参与自发淋巴细胞的产生和改变 收缩，并将继续研究的基础上， 淋巴系统在液体和大分子体内平衡，脂肪 吸收、淋巴细胞循环和抗原呈递。

My long-term career goals are to advance the level of understanding about the roles and mechanisms of an important but poorly understood organ system, the lymphatic system. My immediate career objectives are to: establish a reputation for independent, quality scientific studies; to obtain tenure; to train graduate students and postdoctoral fellows; and to contribute to the advancement of the scientific societies, study section and editorial boards. Receipt of this award will help advance my research program by providing incentives to the college to free up both time and money for the PI to focus on scientific research. The lymphatic system is a specialized part of the circulation that is intimately involved in the maintenance of normal body fluid homeostasis. It is thought to be one of the principal safety factors against gross edema formation. The lymphatic system is also involved in nutrient absorption, lymphocyte circulation and antigen presentation. These tasks are accomplished via lymph circulation. Fluid is actively pumped through the lymphatic system by the spontaneous generation of lymphatic contractions. The general objectives of these studies will be to elucidate the mechanisms involved in the generation and modulation of spontaneous lymphatic contractions. The following specific aims will be addressed: 1) Determine the role of the endothelium, smooth muscle cells and their interactions in the pressure sensitivity of the lymphatic pump; 2) Determine the role of the endothelium, smooth muscle cells and their interactions in the flow sensitivity of the lymphatic pump; 3) Evaluate the regulation of lymphatic intracellular calcium during spontaneous contractions; 4) Evaluate the alterations in lymphatic intracellular calcium during pressure- and flow-induced changes in the lymphatic pump; and 5) Evaluate the site of lymphatic pacemaker activity. These studies will be conducted in isolated, pressurized collecting lymphatics from the mesentery of the rat small intestine. The lymphatics will be dissected out of the tissue and cannulated. The pressure and flow through the vessel will be carefully controlled by varying the inlet and outlet pressures. The role of the endothelium and endothelial-smooth muscle interactions will be investigated using techniques to inhibit and/or remove the lymphatic endothelium. Lymphatic contractions will be monitored using videomicroscopic techniques. The lymphatic diameter will be continuously measured throughout the experiments. From the lymphatic diameter tracings, functional indices of the lymph pump will be evaluated. Intracellular levels of calcium in the lymphatics during spontaneous contractions will be monitored using the fluorescent calcium-sensitive dye Fura-2. The cellular mechanisms involved in the regulation of intracellular calcium will be investigated. The influence of pressure and flow on lymphatic calcium regulation will also be evaluated. Evaluation of membrane potential within the cells of the lymphatic vessels will be performed using the potentiometric fluorescent dye Di-8-ANEPPS. Investigation of the pacemaker activity within the lymphatic will be performed using fluorescent imaging techniques. The results of these investigations should provide considerable insight into the mechanisms involved in the generation and alteration of spontaneous lymphatic contractions and will be the basis for continued studies of the role of the lymphatic system in fluid and macromolecular homeostasis, fat absorption, lymphocyte circulation, and antigen presentation.