Dissecting the preB?tzinger complex neural network using holographic photolysis

描述（由申请人提供）：我的长期目标是了解产生和调节呼吸节律的机制。l将利用一种新的光学方法来激发少量的目标神经元（<10），以解剖前Bvtzinger复合体中的神经微回路，这是产生呼吸节律的关键部位。呼吸是哺乳动物的一种基本的连续行为。调节呼吸的脑干中枢的破坏是睡眠呼吸暂停、雷特综合征、中枢性先天性低通气综合征（CCHS）和可能的婴儿猝死综合征（SIDS）等疾病的基础。神经系统疾病如帕金森氏病、多系统萎缩和肌萎缩侧索硬化症与睡眠呼吸障碍有关。我们目前对呼吸及其突触和电连接的神经元结构的理解是有限的，因此严重阻碍了对健康和疾病中呼吸的神经控制的理解。产生呼吸的吸气相的关键要素是前Bvtzinger复合体（preBvtC），假设其为吸气振荡器的位点。由于preBvtC是由一个异质群体的神经元，并显示没有明显的总体解剖组织，它一直难以区分的preBvtC神经元网络的结构，并确定该结构如何支持呼吸的功能。如果要了解正常和病理条件下的呼吸，必须揭示呼吸节律发生的机制。在这里，从脑干切片包含preBvtzinger复合体和产生呼吸节律，我将研究潜在的机制触发吸气爆发的运动神经活动。利用先进的光学技术，我将同时刺激一到几个前Bvtzinger复合体吸气神经元。我提供了初步的数据，3-9 preBvtzinger复杂吸气神经元的激活可以触发吸气运动神经爆发。我建议验证，完善和扩展这些实验，以阐明preBvtC的结构和功能，控制呼吸的神经回路的关键要素。通过利用先进的光学方法，我可以实现我的最终目标，即获得足够的数据集，用于代表性和可测试的建模，其中包含详细的preBvtC电路拓扑结构，这是深入了解这种重要行为所必需的。这里获得的数据有可能为理解呼吸节律机制提供一个非凡的窗口。 公共卫生相关性：前Bvtzinger复合体（preBvtC）是呼吸基础脑干回路的重要组成部分。preBvtC的破坏被认为是睡眠呼吸暂停、Rett综合征和婴儿猝死综合征等疾病的基础，并与帕金森病、多系统萎缩和肌萎缩性侧索硬化等神经系统疾病中的睡眠呼吸障碍有关。preBvtC网络结构的知识将极大地帮助我们理解呼吸是如何产生的，并进一步帮助我们解决呼吸中枢中断后的健康相关问题。

DESCRIPTION (provided by applicant): My long-term objective is to understand the mechanisms generating and modulating respiratory rhythm. l will exploit a novel optical method for exciting small numbers of targeted neurons (<10) to dissect the neural microcircuit in the preBvtzinger Complex, a key site for generation of respiratory rhythm. Breathing is an essential continuous behavior in mammals. Disruption of brainstem centers regulating breathing underlies disorders such as sleep apnea, Rett syndrome, central congenital hypoventilation syndrome (CCHS) and possibly sudden infant death syndrome (SIDS). Neurologic disorders such as Parkington's disease, multiple systems atrophy and amyotropic lateral sclerosis are associated with sleep- disordered breathing. Our current understanding of the neuronal structures underlying respiration and their synaptic and electrical connectivity is limited, and as such a serious hindrance to understanding the neural control of breathing in health and disease. A critical element for generation of the inspiratory phase of respiration is the preBvtzinger Complex (preBvtC), hypothesized to be the site of the inspiratory oscillator. Because the preBvtC is comprised of a heterogeneous population of neurons, and displays no obvious gross anatomical organization, it has been difficult to distinguish the structure of the preBvtC neuronal network and to determine how this structure supports the function of respiration. If breathing is to be understood in normal and in pathological conditions, the mechanisms for respiratory rhythmogenesis must be revealed. Here, in slices from the brainstem that contain the preBvtzinger Complex and generate a respiratory rhythm, I will study the mechanisms underlying the triggering of inspiratory bursts of motor nerve activity. Using advanced optical techniques, I will simultaneously stimulate one to several preBvtzinger Complex inspiratory neurons. I provide preliminary data that activation of 3-9 preBvtzinger Complex inspiratory neurons can trigger an inspiratory motor nerve burst. I propose to validate, refine and extend these experiments to elucidate the structure and function of preBvtC, a key element of the neural circuit controlling breathing. By exploiting advanced optical methods, I can achieve my ultimate goal of obtaining a dataset sufficient for representative and testable modeling that incorporates detailed preBvtC circuit topology, necessary for a deep understanding of this vital behavior. The data obtained here has the potential to provide an extraordinary window into understanding mechanisms of respiratory rhythm. PUBLIC HEALTH RELEVANCE: The preBvtzinger Complex (preBvtC) is an essential element of the brainstem circuitry underlying breathing. Disruption of the preBvtC is proposed to underlie disorders such as sleep apnea, Rett syndrome, and sudden infant death syndrome and has been associated with sleep-disordered breathing in neurologic disorders such as Parkinson's disease, multiple system atrophy, and amyotropic lateral sclerosis. Knowledge of the preBvtC network structure will greatly aid our understanding of how respiration is generated and further our efforts to tackle the health-related issues that follow central disruption of breathing.