Interactions of the Noradrenergic and Serotonergic Systems in Autoresuscitation

项目摘要/摘要 脑干的去甲肾上腺素(NA)和5-羟色胺(5-HT)网络是高度调节和必需的 呼吸动态平衡网络的组成部分。这些系统的功能障碍与许多 先天性呼吸系统疾病，包括婴儿猝死综合症(SIDS)。小岛屿发展中国家是导致 新生儿死亡，被认为部分是由于新生儿自我复苏反射的失败。上一首 数据让我们了解了这些系统如何调节对生命的保护性呼吸反应 独立威胁挑战，但关于两国之间潜在相互作用的信息有限 这两个关键系统虽然以前的研究缺乏一个更巧妙的操作，我们的方法论。 在以下目标中，我将检验5-羟色胺和NA系统在调节中是完整联系的假设 自动复苏反射以及5-羟色胺和去甲肾上腺素不同组合扰动和刺激将 导致自动复苏的阳性和阴性结果相加。目标1.确定独立的和 5-羟色胺能激活和去甲肾上腺素能抑制的联合功能和细胞效应 自主复苏反射和呼吸网络动力学对高碳酸血症的反应。目标2.确定 5-羟色胺能抑制和去甲肾上腺素能激活对自复苏反射的影响。目标3. 测定5-羟色胺能抑制和去甲肾上腺素能抑制或5-羟色胺能激活的效果 去甲肾上腺素对自动复苏反射的激活作用。为了测试这些目标，我将使用复合 重组和效应线，以访问和操纵5-羟色胺和NA系统的活动。例如, 将Pet1：：Cre；F_hM4D系培育成DBH_FLPO；P_hM3D系将产生小鼠，对氯氮平-N-氧化物 (CNO)的应用将同时兴奋5-羟色胺系统(Pet1：：Cre+P_hM3D)，同时抑制NA系统 (DBH_FLPO+F_hM4D)。对于功能特征，小鼠将被反复发作的挑战 来测试他们的自动复苏反射。对于网络活动表征，CFOS染色将 对亚致死性高二氧化碳缺氧小鼠的脑干组织进行了研究 5-羟色胺或去甲肾上腺素系统兴奋或抑制后暴露。了解这些系统是否 独立调节自动复苏将为未来的治疗研究提供有价值的信息。 这项研究将通过扩大我们对5-羟色胺和NA系统之间相互作用的理解来推动这一领域的发展 在健康和疾病方面，以及在这两个研究中使用复杂的基因操作的信息 关键的呼吸系统。这项工作将极大地促进我们对所涉及的神经网络的理解 在呼吸调节和疾病方面。本建议书附带的培训计划和环境将 为向研究独立过渡提供坚实的基础。培训包括科学性和专业性 开发课程、研讨会和研讨会，而环境包括尖端技术和 在BCM提供内部培训的设备。

Project Summary/ Abstract The noradrenergic (NA) and serotonergic (5-HT) networks of the brainstem are highly regulated and necessary components of the respiratory homeostatic network. Dysfunction of these systems has been linked to many congenital respiratory disorders including Sudden Infant Death Syndrome (SIDS). SIDS is a leading cause of neonate death and is thought to occur, in part, by the failure of the neonate autoresuscitation reflex. Previous data have informed our understanding of how these systems modulate protective respiratory responses to life threatening challenges independently, but limited information is available on the potential interplay between these two key systems while previous studies lack a more finessed manipulation provided by our methodology. In the following aims, I will test the hypothesis that the 5-HT and NA systems are integrally linked in regulating the autoresuscitation reflex and that different combinations of 5-HT and NA perturbation and stimulation will result in additive positive and negative outcomes in autoresuscitation. Aim 1. Determine the separate and combined functional and cellular effects of serotonergic activation and noradrenergic inhibition on the autoresuscitation reflex and respiratory network dynamics in response to hypercapnic anoxia. Aim 2. Determine the effect of serotonergic inhibition and noradrenergic activation on the autoresuscitation reflex. Aim 3. Determine the effect of serotonergic inhibition and noradrenergic inhibition or serotonergic activation and noradrenergic activation on the autoresuscitation reflex. To test these aims, I will utilize compounded recombinase and effector lines to access and manipulate the activity of the 5-HT and NA systems. For example, the Pet1::Cre; F_hM4D line bred to a DBH_FLPo; P_hM3D line will produce mice that, upon clozapine-N-oxide (CNO) application, will concurrently excite the 5-HT system (Pet1::Cre + P_hM3D) while inhibiting the NA system (DBH_FLPo + F_hM4D). For functional characterizations, the mice will be challenged with repeated bouts of hypercapnic anoxia to test their autoresuscitation reflex. For network activity characterization, cFos staining will be carried out on brainstem tissue in mice that have been challenged with sublethal hypercapnic anoxic exposures after excitation or inhibition of the 5-HT or NA systems. Understanding if these systems are independently regulating autoresuscitation will provide valuable information for future therapeutic investigations. This study will progress the field by expanding our understanding of interplay between the 5-HT and NA systems in health and disease as well as informing on the use of intricate genetic manipulations in the study of these two key respiratory systems. This work will significantly advance our understanding of the neural networks involved in respiratory regulation and disease. The training plan and environment that accompany this proposal will provide a solid foundation for a transition to research independence. Training includes scientific and professional development courses, workshops, and seminars while the environment includes cutting edge technology and equipment with training offered in house at BCM.