Na+ TRANSPORT INHIBITION BY RESPIRATORY SYNCYTIAL VIRUS

应聘者：我1992年毕业于布里斯托尔大学，获解剖学和兽医学荣誉学位。在UAB，我接受了实验动物医学、实验动物病理学和动物模型病毒发病机制方面的培训(2000年2月博士；导师Patricia Fultz博士)。随后，我选择加入马塔隆博士的研究小组，在那里我可能会通过电生理学交叉挖掘我对病毒发病机制的了解，并将这些技术应用于研究人类呼吸道病毒肺炎的发病机制。 研究：呼吸道合胞病毒(RSV)是全球儿童下呼吸道疾病的最常见原因。RSV诱导的毛细支气管炎的发病机制尚不清楚，RSV感染对离子转运(呼吸道上皮细胞的一种精液功能)的影响尚未被研究。我推测呼吸道合胞病毒感染呼吸道上皮细胞会降低它们的Na？运输能力。初步研究表明，无论是在体外还是在感染RSV的BALB/c小鼠中，这一假设都是正确的。我01年的目标- 这些研究包括：(1)定量研究呼吸道合胞病毒感染BALB/c小鼠后，体内外小鼠呼吸道和肺泡上皮细胞钠离子转运的变化；(2)确定呼吸道合胞病毒感染体外培养的小鼠上皮细胞后，钠电流和阿米洛利敏感通道活性的变化；以及(3)研究呼吸道合胞病毒诱导的体内外Na+转运的变化与海洋呼吸上皮细胞ENaC表达的变化之间的关系。我在04-05年的计划是确定泛素/蛋白酶体途径在呼吸道上皮细胞感染RSV后调节Na+转运减少中的作用，并确定RSV小疏水(SH)基因产物在调节Na+转运中的作用。我会用电生理和生化相结合的方法 目的探讨呼吸道合胞病毒(RSV)对Na+转运的影响，并探讨其与Na+通道表达和降解的关系。我的项目将强调不同技术的交叉培训，包括跨单分子层的短路测量、放射性同位素离子通量研究、全细胞和单通道膜片钳，以及测量小鼠的肺泡液清除和鼻电位差。 环境：这个由Matalon博士(肺生理学)和Sullender博士(呼吸道合胞病毒)分别担任导师和共同导师的SERCA，将为我提供所需的培训和环境，使我成为一名独立科学家，专注于比较呼吸道病毒对正常上皮细胞功能的病理生理影响。

CANDIDATE: I graduated from the University of Bristol in 1992, with Honours in Anatomical Science and Veterinary Science. At UAB I received training in laboratory animal medicine, laboratory animal pathology, and viral pathogenesis in animal models (Ph.D. February 2000; mentor Dr. Patricia Fultz). Subsequently, I elected to join Dr. Matalon's research group, where I might build on my knowledge of viral pathogenesis by cross-mining in electrophysiology, and apply these techniques to study the pathogenesis of human respiratory viral pneumonitides. RESEARCH: Respiratory syncytial virus (RSV) is commonest cause of lower respiratory tract disease in children worldwide. Pathogenesis of RSV-induced bronchiolitis is poorly understood, and effects of RSV infection on ion transport (a seminal function of respiratory epithelial cells) have not been investigated. I hypothesize that RSV infection of respiratory epithelial cells reduces their Na ? transport capacity. Preliminary studies have demonstrated that this hypothesis is correct, both in vitro and in RSV-infeeted BALB/c mice. My aims for years 01- 03 are to: (1) quantify alterations in Na + transport across airway and alveolar epithelia in vivo and ex vivo, after infection of BALB/c mice with RSV; (2) define changes in Na + currents and amiloride-sensitive channel activity after RSV infection of murine epithelial cells in vitro; and (3) correlate alterations in Na + transport induced by RSV in vitro and in vivo with alterations in ENaC expression by marine respiratory epithelia. My plan for years 04-05 is to determine the role of the ubiquitin/proteasome pathway in mediating reduced Na+ transport after RSV infection of respiratory epithelia, and to identify the role of the RSV small hydrophobic (SH) gene product in modulation of Na + transport. I will use a combined electrophysiologic and biochemical approach to investigate effects of RSV on Na + transport at all levels, from the single cell to the whole animal, and to correlate these effects to Na + channel expression and degradation. My project will emphasize cross-training in diverse techniques, including short-circuit measurements across monolayers, radioisotopic ion flux studies, whole cell and single channel patch-clamp, and measurement of alveolar fluid clearance and nasal potential difference in mice. ENVIRONMENT: This SERCA, with Dr. Matalon (lung physiology) as mentor and Dr. Sullender (respiratory syncytial virus) as co-mentor, will provide the training and setting I require for my maturation into an independent scientist focused on comparative pathophysiologic effects of respiratory viruses on normal epithelial cell function.