DEVELOPMENTAL REGULATION OF OXYGEN SENSING IN THE LUNG

肺氧传感的发育调节

• 批准号: 6041739
• 负责人: DAVID N. CORNFIELD
• 金额: $ 28.72万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6041739
• 关键词: amphotericin B; calcium ion; confocal scanning microscopy; congenital cardiovascular disorder; embryo /fetus cell /tissue; immunocytochemistry; lung; membrane potentials; newborn animals; oxygen; oxygen tension; oxygen therapy; perinatal; potassium channel; pulmonary circulation; pulmonary hypertension; sheep; smooth muscle; tissue /cell culture; vasoconstriction; vertebrate embryology; voltage /patch clamp

At birth, the pulmonary circulation changes dramatically. Pulmonary blood flow increases 8-10 fold while pulmonary arterial (PA) pressure declines. If the pulmonary circulation does not make the transition to a high flow, low pressure circuit, then persistent pulmonary hypertension of the newborn (PPHN) results. PPHN is characterized by increased pulmonary vascular tone and reactivity, resulting in severe central hypoxemia that responds incompletely to the administration of supplemental O2 and other vasodilator stimuli. Neither prevention nor cure is currently available. Treatment options are compromised by incomplete understanding of the mechanisms responsible for postnatal adaptation of the pulmonary circulation. While the acute increase in O2 tension is clearly an essential physiologic stimulus for pulmonary vasodilation in the perinatal period, the identity of the O2 sensor remains unknown. Thus, the mechanism whereby the perinatal pulmonary vasculature senses and vasodilates in response to the acute increase in O2 tension remains incompletely understood. Treatment strategies are compromised by the lack of molecular targets for therapy. This proposal seeks to determine whether the ability of the pulmonary vasculature to respond to an acute increase in O2 tension is developmentally regulated and the subcellular mechanisms that allow the perinatal pulmonary circulation to vasodilate in response to an increase in O2 tension. Evidence suggests that potassium (K+) channels in PA smooth muscle cells (SMC) act as the O2 sensor. Work from our laboratory suggests that the calcium-sensitive K+ (KCa) channel plays a central role in mediating the response of the perinatal pulmonary circulation to an acute increase in O2 tension. We propose the working HYPOTHESIS that oxygen activates a KCa channel through intracellular calcium release to cause PA SMC Em hyperpolarization, contributing to perinatal pulmonary vasodilation. The specific aims are to test the following hypotheses: Aim 1. The KCa channel is a major component of PA SMC O2 sensing in the late-gestation fetus and newborn and is developmentally regulated Aim 2. Activation of the PA SMC KCa channel through Ca2+ sparks contributes to perinatal pulmonary vasodilation. To test these hypotheses, we plan experiments using electrophysiology, dynamic calcium imaging, confocal microscopy, molecular biology, immunocytochemistry, and whole animal physiology. The experiments will provide definitive identification of an O2 sensor in the neonatal pulmonary circulation that may be developmentally regulated and elucidate the subcellular processes that lead to its activation, thereby providing a novel molecular target to address neonatal pulmonary vascular disease.

出生时，肺循环发生了巨大的变化。肺血流量增加8-10倍，而肺动脉（PA）压力下降。 如果肺循环没有过渡到高流量、低压回路，则会导致新生儿持续性肺动脉高压（PPHN）。PPHN的特征是肺血管张力和反应性增加，导致严重的中枢低氧血症，对补充O2和其他血管扩张刺激的给药反应不完全。 目前既没有预防也没有治疗。 由于对肺循环出生后适应机制的不完全理解，治疗方案受到影响。 虽然氧张力的急性增加显然是围产期肺血管舒张的重要生理刺激，但氧传感器的身份仍然未知。 因此，围产期肺血管系统的感觉和血管舒张的机制，以响应急性增加的O2张力仍然不完全清楚。 治疗策略因缺乏治疗的分子靶点而受到影响。 该提案旨在确定肺血管系统对O2张力急性增加的反应能力是否受到发育调节，以及允许围产期肺循环对O2张力增加作出反应而血管舒张的亚细胞机制。 有证据表明，PA平滑肌细胞（SMC）的钾（K+）通道作为O2传感器。 我们实验室的工作表明，钙敏感性K+（KCa）通道在介导围产期肺循环对O2张力急性增加的反应中起着核心作用。 我们提出的工作假设，氧激活KCa通道，通过细胞内钙释放，导致PA SMC Em超极化，有助于围产期肺血管舒张。 具体目标是检验以下假设：目标1。KCa通道是妊娠晚期胎儿和新生儿PA SMC O2感知的主要组成部分，并受发育调节。 通过Ca 2+火花激活PA SMC KCa通道有助于围产期肺血管舒张。 为了验证这些假设，我们计划使用电生理学，动态钙成像，共聚焦显微镜，分子生物学，免疫细胞化学和整个动物生理学实验。 这些实验将提供新生儿肺循环中O2传感器的明确鉴定，该传感器可能受到发育调节，并阐明导致其激活的亚细胞过程，从而提供一种新的分子靶点来解决新生儿肺血管疾病。