Exploring the function of the central control of breathing in mice with sodium-channel mutations causing epilepsy, implication for the sudden unexpected death in epilepsy (SUDEP).

呼吸对生命至关重要，控制这种行为的神经元网络功能障碍被怀疑与几种神经系统疾病有关。前波青格复合体（PreBötC）是位于脑干的吸气呼吸活动的中枢发生器。这项研究计划旨在揭示编码电压门控钠通道的基因中已知的致癫痫突变对PreBötC功能的影响。该项目旨在研究这些基因中靶向敲除和敲入突变的小鼠体内和体外呼吸活动及其对SUDEP的可能后果。癫痫猝死（SUDEP）是癫痫患者最常见的直接死亡原因。具体而言，将检查基因SCN1A和SCN2A突变的小鼠。首先，将非常详细地表征突变小鼠的呼吸，并与野生型（WT）同窝仔进行比较。我建议使用自由活动小鼠的体内全身体积描记法记录来表征呼吸参数（目标1）。其次，我将利用从含有PreBötC的脑干切片中获得的体外电生理学和组织学，在网络和细胞水平上剖析这些突变引起的可能的病理机制（目标2）。在我以前的工作中，我已经展示了单基因突变（CACNA1A和NDUFS4，初步工作）如何导致呼吸中枢控制的严重和潜在的致命功能障碍。 目标1和2的结果不仅将提供对这些基因突变如何导致病理的理解，而且将增强我们对呼吸中枢控制的基本功能的认识。此外，在第三组实验中，我建议研究抗癫痫药物（AED）对野生型和突变小鼠体内和体外PreBötC功能的影响（目标3），以了解这些药物如何影响PreBötC的功能。导致SUDEP的确切病理生理机制尚未完全了解。怀疑SUDEP涉及心脏跳动和呼吸的自主控制功能障碍。因此，PreBötC的功能可能在导致这些患者死亡的致命级联反应中发挥关键作用。第一，第二和第三系列实验的结果将相互关联并结合起来，以揭示PreBötC的失调是否以及如何参与这些事件。

Breathing is essential for life and a dysfunction of the neuronal network that controls this behaviour is suspected to be involved in several neurological diseases. The PreBötzinger Complex (PreBötC) is the central generator of the inspiratory breathing activity located in the brainstem. This research proposal is designed to unravel the consequences, of known epilepsy-causing mutations in genes encoding voltage gated sodium-channels, on the function of the PreBötC. The project aims to investigate the breathing activity in vivo and in vitro in mice with targeted knock-out and knock-in mutations in these genes and their possible consequences for SUDEP. Sudden unexpected death in epilepsy (SUDEP) is the most common direct cause of death in people suffering from epilepsy. Specifically, mice with mutations in the genes SCN1A and SCN2A will be examined. First, breathing will be characterized in great detail in the mutant mice and compared to the wildtype (WT) littermates. I propose to use in vivo whole body plethysmography recordings of freely moving mice to characterize the breathing parameters (objective1). Second, I will utilize in vitro electrophysiology and histology, obtained from brainstem slices containing the PreBötC, to dissect the possible pathological mechanisms at the network and cellular level caused by these mutations (objective 2). In my previous work, I have already shown how monogenic mutations (CACNA1A and NDUFS4, preliminary work) can lead to severe and potential lethal dysfunction of the central control of breathing. The results of objective 1 and 2 will provide, not only, an understanding how mutations in these genes might cause the pathology, but will enhance our knowledge of the fundamental function of the central control of breathing. Furthermore, in a third set of experiments, I propose to investigate the effects of Antiepileptic drugs (AED) on the function of the PreBötC in vitro and in vivo (objective 3) in wildtype and mutant mice to understand how these drugs might affect the function of the PreBötC. The exact pathophysiology leading to SUDEP is not well understood. A dysfunction of the autonomic control of heart beat and breathing are suspected to be involved in SUDEP. The function of the PreBötC might therefore play a crucial role in the fatal cascade leading to the death in these patients. The results of the first, second and third series of experiments will be correlated and combined to unravel if and how a dysregulation of the PreBötC could be involved in these events.