MECHANISMS INVOLVED IN LYMPHATIC CONTRACTILE ACTIVITY

我的长期职业目标是提高对 一个重要但鲜为人知的器官的作用和机制 系统，淋巴系统。我目前的职业目标是： 建立独立、高质量的科学研究的声誉； 获得终身教职；培训研究生和博士后研究员；以及 为科学学会的进步作出贡献，研究部分 和编辑委员会。获得这个奖项将有助于推动我的研究 计划通过向学院提供激励来腾出时间和 为PI专注于科学研究提供资金。淋巴系统是 循环的一个专门部分，密切地参与 维持正常的体液平衡。它被认为是 防止肉眼水肿形成的主要安全因素。淋巴管 系统还参与营养吸收、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。 液体被自发地通过淋巴系统泵入 淋巴收缩的产生。这些活动的总体目标是 研究将阐明发生和发生的机制 自发性淋巴收缩的调节。以下是具体的 目标将解决：1)确定内皮细胞的作用，顺畅 肌细胞及其相互作用在人体压力敏感性中的作用 淋巴泵；2)测定血管内皮细胞、平滑肌的作用 淋巴管流动敏感性中的细胞及其相互作用 泵；3)评价淋巴细胞内钙的调节。 自发收缩；4)评估淋巴管的变化 压力和流量诱导的细胞内钙离子变化 淋巴泵；5)评价淋巴起搏器活动的部位。 这些研究将在隔离的加压收集中进行 来自大鼠小肠系膜的淋巴管。淋巴管 将被从组织中解剖出来并插入管子。压力和流量 通过改变进水口和进水口将仔细控制通过该容器 出口压力。血管内皮细胞和血管内皮细胞的作用 肌肉的相互作用将使用抑制技术进行研究 和/或去除淋巴管内皮细胞。淋巴收缩将是 使用视频显微镜技术进行监测。淋巴管直径将会 在整个实验过程中被连续测量。来自淋巴管 对淋巴泵的直径、功能指标进行评估。 自发性淋巴管细胞内钙离子水平 宫缩将使用荧光钙敏感染料进行监测 富拉-2。参与调控的细胞机制 将对细胞内钙离子进行研究。压力和压力的影响 还将评估血流对淋巴钙调节的影响。评估 淋巴管细胞内的膜电位 使用电位荧光染料Di-8-ANEPPS。 对淋巴管内起搏器活动的研究将 使用荧光成像技术进行。这些研究的结果 调查应该能为这些机制提供相当大的洞察力 参与自发性淋巴管的产生和改变 宫缩，并将成为继续研究的基础 液体和大分子动态平衡中的淋巴系统，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.