Modulation of calcium induced calcium release in adrenergic neuron via nNOS nerve

通过 nNOS 神经调节肾上腺素能神经元中钙诱导的钙释放

• 批准号: 7541404
• 负责人: J L BUCHHOLZ
• 金额: $ 19.31万
• 依托单位: LOMA LINDA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7541404
• 关键词: ATP phosphohydrolase; Acclimatization; Adrenergic Agents; Adult; Agonist; Altitude; Appendix; Attenuated; Blood Vessels; Blood flow; Caffeine; Calcium; Cerebrovascular Circulation; Cerebrum; Citrulline; Communication; Complex; Condition; Cyclic ADP-Ribose; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; Cytophotometry; Data; Development; Electric Stimulation; Elevation; Fetus; Flavin Mononucleotide; Genetic; Goals; Guanylate Cyclase; Health; Human; Hypertension; Hypoxemia; Hypoxia; KT 5823; Measures; Mediating; Messenger RNA; Methods; Modeling; Molecular; Nerve; Neurons; Nicotinamide adenine dinucleotide; Nitric Oxide; Nitric Oxide Donors; Nitric Oxide Synthase Type I; Pathology; Pathway interactions; Patient currently pregnant; Penicillamine; Polymerase; Process; Proteins; Protocols documentation; Publishing; Pump; Range; Rattus; Reaction; Regulation; Reverse Transcription; Risk; Severities; Sheep; Signal Pathway; Signal Transduction; Smooth Endoplasmic Reticulum; Stress; Stroke; Structure of superior cervical ganglion; Study models; Techniques; Time; Western Blotting; Work; adrenergic; analog; fetal; ganglion cell; indexing; liquid chromatography mass spectroscopy; middle cerebral artery; programs; response; systolic hypertension

This proposal is integral to the overall PPG theme to study fetal and adult adaptations to long-term hypoxia (LTH) at high altitude. Protection is needed against cerebral insults not only in the adult but also in the fetus exposed to systolic hypertension during LTH. Adrenergic neurons, arising from the superior cervical ganglion (SCG), are an important component of the regulation of cerebral blood vessel contractility and blood flow under this stress. Furthermore, these neurons regulate cerebral blood flow under stress conditions such as hypoxemia and hypertension, reducing the risk of stroke, and attenuating severity of pathology following subarachnoid hemmorhage. Our earlier published data show that adrenergic nerve function in middle cerebral arteries (MCA) is facilitated via neuronal nitric oxide synthase (nNOS) containing nerves co-innervating the cerebral vasculature. These data suggest that there is a complex communication pathway between these two nerve types. We now are focusing on the SCG as it is a well established and efficient model for studying the complex processes that control intracellular calcium ([Ca2+]i) in adrenergic nerves and modulation by nNOS nerves. The function of adrenergic nerves depends in part on calcium induced calcium release (CICR) from the smooth endoplasmic reticulum (SER). CICR amplifies increased [Ca2+]i caused by influx through Ca2+ channels and requires that SER be filled by Ca 2+ influx through store operated Ca2+ channels (SOCC) and SER Ca 2+-ATPases (SERCAs). The faciliatory function of nNOS nerves on adrenergic nerves in the SCG may, in part, be due to amplification of the CICR process. We have shown that the function of faciliatory nNOS nerves declines during LTH at high altitude, which is partly related to a decline in nNOS protein levels. These observations may have implications for the distribution of blood flow during LTH acclimatization at high altitude. Studying mechanisms underlying the regulation of adrenergic nerves is vital to human health and development -- from fetus to adult. The project goal is to determine mechanisms underlying the impact of LTH and development on intracellular calcium ([Ca2+]) signaling in adrenergic neurons arising from the SCG and modulation of these processes by nNOS nerves. The governing hypothesis is: positive modulation of adrenergic nerves via nNOS nerves through the CICR process or refilling of SER Ca 2+ stores declines in response to LTH, We will use a range of techniques, instrumental, pharmacological and molecular to probe the mechanisms by which LTH stress may alter the refilling of SER Ca 2+ stores and/or the signaling pathway responsible for modulating CICR in isolated sheep SCG cells. The four groups to be studied are: adult non-pregnant normoxic and high altitude acclimated, and near term fetus, normoxic and high altitude.

该提案是 PPG 总体主题的一部分，旨在研究胎儿和成人对高海拔长期缺氧 (LTH) 的适应。不仅成人需要保护脑损伤，而且在 LTH 期间暴露于收缩期高血压的胎儿也需要保护脑损伤。由颈上神经节（SCG）产生的肾上腺素能神经元是在这种应激下调节脑血管收缩性和血流的重要组成部分。此外，这些神经元在低氧血症和高血压等应激条件下调节脑血流，降低中风风险，并减轻蛛网膜下腔出血后病理的严重程度。我们之前发表的数据表明，中脑的肾上腺素能神经功能 动脉（MCA）是通过神经元一氧化氮合酶（nNOS）促进的，其中神经元一氧化氮合酶含有共同神经支配脑血管系统的神经。这些数据表明这两种神经类型之间存在复杂的通讯途径。我们现在关注 SCG，因为它是一个成熟且有效的模型，用于研究控制肾上腺素能神经中细胞内钙 ([Ca2+]i) 和 nNOS 神经调节的复杂过程。肾上腺素能神经的功能部分取决于钙诱导的钙释放（CICR） 平滑内质网（SER）。 CICR 放大由 Ca2+ 通道流入引起的 [Ca2+]i 增加，并要求 SER 由通过钙池操纵 Ca2+ 通道 (SOCC) 和 SER Ca 2+ -ATP 酶 (SERCA) 流入的 Ca 2+ 填充。 nNOS 神经对 SCG 中肾上腺素能神经的促进功能可能部分归因于 CICR 过程的放大。我们发现，在高海拔 LTH 期间，促进 nNOS 神经的功能下降，这部分与 nNOS 蛋白水平下降有关。这些观察结果可能对高海拔 LTH 适应过程中的血流分布有影响。学习中 肾上腺素能神经调节的机制对于人类从胎儿到成人的健康和发育至关重要。该项目的目标是确定 LTH 和发育对 SCG 产生的肾上腺素能神经元中细胞内钙 ([Ca2+]) 信号传导的影响机制以及 nNOS 神经对这些过程的调节。主要假设是：肾上腺素能神经通过 nNOS 神经通过 CICR 过程或 SER Ca 2+ 储存的再填充因 LTH 而下降，我们将使用一系列技术、仪器、药理学和分子来探讨 LTH 应激可能改变 SER Ca 2+ 储存的再填充和/或负责调节离体绵羊 SCG 细胞 CICR 的信号通路。要研究的四个组是：常氧和高海拔适应的成人非妊娠组，以及常氧和高海拔的近足月胎儿。