Enhanced respiratory plasticity in models of respiratory motor neuron death

项目总结/摘要 候选人和环境：候选人的近期和长期目标是获得知识， 成为一名成功的学术研究人员所需的经验，专注于独立的实验室 呼吸的神经控制威斯康星大学麦迪逊分校有许多世界知名的呼吸生理学家，满足 每周为呼吸神经生物学研讨会系列和杂志俱乐部系列，使其成为一个独特的研究 在这样的环境中，候选人可以茁壮成长。因此，留在威斯康星大学麦迪逊分校[将使候选人， 由神经可塑性和呼吸神经生物学的顶尖研究人员指导（这是为数不多的地方之一）。 学习这种组合），并获得所需的知识和经验，过渡到独立。 研究：ALS是一种破坏性疾病，导致进行性运动神经元变性和受损 呼吸，最终导致死亡。尽管呼吸功能不全非常重要， 很少在任何ALS模型中进行研究。因此，需要采取策略来保持足够的排泄功能 来保护生命。这项建议的基本目标是确定补偿机制， 增强的呼吸可塑性（间歇性缺氧诱导的膈神经长时程易化; pLTF）。增强 可塑性可能使我们能够在呼吸运动神经元死亡的模型中恢复膈运动输出，从而 保留了解释能力。我们建议在大鼠呼吸运动神经元死亡模型中研究pLTF 包括ALS遗传模型（SOD 1G 93 A大鼠）中遇到的进行性疾病和稳定的， 诱导型模型（缀合至皂草素的霍乱毒素B; CTB-SAP）。我们建议使用多学科 方法包括：1）使用CTB-SAP诱导呼吸运动神经元死亡; 2）膈神经记录， 直接评估运动输出; 3）体内RNAi以评估CTB-SAP中AIH诱导的pLTF的细胞机制 免疫组化方法测定膈运动神经元存活及其表达， 关键分子[3]提出了具体的目标：1）增加SOD 1G 93 A表达触发代偿性 NADPH氧化酶表达增加，从而保持足以表达pLTF的ROS水平; [2] 诱导的呼吸运动神经元死亡增强pLTF;和3）诱导的呼吸运动神经元死亡 通过与SOD 1G 93 A大鼠不同的机制增强pLTF。由于大多数ALS患者 呼吸功能不全，导致呼吸机依赖或死亡，我们的长期目标是开发新的 延缓呼吸运动神经元死亡和增强备用运动神经元功能的策略 神经元在这里，我们将利用机制，增加存活的运动神经元（AIH诱导的）的功能， 可塑性）。这些新的策略，如果成功的话，可能会指导未来的转化研究， 呼吸运动神经元死亡，包括ALS患者。目前的研究可能具有广泛的影响， 有益于ALS和其他神经退行性疾病的非呼吸性运动池。

Project Summary/Abstract Candidate and Environment: The immediate and long-term goals of the candidate are to gain knowledge and experience necessary to become a successful academic researcher with an independent laboratory focused on the neural control of breathing. UW-Madison has many world-renowned respiratory physiologists that meet weekly for a respiratory neurobiology seminar series and journal club series making it a unique research environment in which the candidate can thrive. Thus, remaining at UW-Madison [will enable the candidate to be mentored by top-notch researchers in neuroplasticity and respiratory neurobiology (one of very few places studying this combination) and attain the required knowledge and experience to transition to independence.] Research: ALS is a devastating disease leading to progressive motor neuron degeneration and compromised breathing which ultimately leads to death. Despite its fundamental importance, respiratory insufficiency has seldom been studied in any ALS model. Thus, strategies to preserve adequate ventilatory function are needed to preserve life. The fundamental goal of this proposal is to identify compensatory mechanisms that trigger enhanced respiratory plasticity (intermittent hypoxia induced phrenic long-term facilitation; pLTF). Enhanced plasticity may allow us to restore phrenic motor output in models of respiratory motor neuron death, thereby preserving ventilatory capacity. We propose to study pLTF in rat models of respiratory motor neuron death including the progressive disease encountered in a genetic model of ALS (SOD1G93A rats) and a stable, inducible model (cholera toxin B conjugated to saporin; CTB-SAP). We propose to use a multidisciplinary approach including: 1) inducing respiratory motor neuron death using CTB-SAP; 2) phrenic nerve recordings to directly assess motor output; 3) RNAi in vivo to assess cellular mechanisms of AIH-induced pLTF in CTB-SAP rats; and 4) immunohistochemical methods to determine phrenic motor neuron survival and their expression of key molecules. [Three] specific aims are proposed: 1) increased SOD1G93A expression triggers compensatory increases in NADPH oxidase expression, thereby preserving ROS levels sufficient to express pLTF; [2) induced respiratory motor neuron death enhances pLTF; and 3) induced respiratory motor neuron death enhances pLTF by a mechanism distinct from that in SOD1G93A rats.] Since most ALS patients develop respiratory insufficiency, leading to ventilator dependence or death, our long-range goal is to develop new strategies to delay respiratory motor neuron death and enhance the functional capacity of spared motor neurons. Here we will harness mechanisms that increase function of surviving motor neurons (AIH-induced plasticity). These novel strategies, if successful, may guide future, translational studies in patients suffering from respiratory motor neuron death, including ALS patients. The present studies may have wide reaching benefits for non-respiratory motor pools in ALS, and other neurodegenerative diseases.