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奏的机制。我将利用一种新的光学方法来激发少量的目标神经元（<10）来解剖preBvtzinger Complex中的神经微电路，preBvtzinger Complex是产生呼吸节律的关键部位。呼吸是哺乳动物必不可少的连续行为。调节呼吸的脑干中枢的破坏是睡眠呼吸暂停、Rett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）等疾病的潜在原因。神经系统疾病如帕金顿氏病、多系统萎缩和肌萎缩性侧索硬化症与睡眠呼吸障碍有关。我们目前对呼吸的神经元结构及其突触和电连接的理解是有限的，这严重阻碍了我们对健康和疾病中呼吸的神经控制的理解。呼吸吸气相产生的一个关键因素是preBvtzinger复合体（preBvtC），它被假设为吸气振荡器的位置。由于preBvtC由异质的神经元群组成，并且没有明显的大体解剖组织，因此很难区分preBvtC神经元网络的结构并确定该结构如何支持呼吸功能。如果要理解正常和病理状态下的呼吸，就必须揭示呼吸节律发生的机制。这里，在脑干的切片中包含preBvtzinger复合体并产生呼吸节律，我将研究触发运动神经活动的吸气爆发的机制。利用先进的光学技术，我将同时刺激一到几个普瑞布青格复合体的吸气神经元。我提供的初步数据表明3-9个preBvtzinger复合体吸气神经元的激活可以引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，preBvtC是控制呼吸的神经回路的关键元素。通过利用先进的光学方法，我可以实现我的最终目标，即获得一个足以进行代表性和可测试建模的数据集，该数据集包含详细的preBvtC电路拓扑，这对于深入理解这一重要行为是必要的。这里获得的数据有可能为了解呼吸节律的机制提供一个非凡的窗口。

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.