GABAergic Inhibition and the Ketogenic Diet

描述(由申请者提供)：本申请是关于独立科学家奖(K02)的申请。候选人是一位对儿童癫痫有特殊兴趣的儿科神经科医生，目前是K08(MCSDA)资助的最后一年。他最近调到了加州大学欧文分校(UCI)。UCI拥有国际公认的基础神经科学研究社区，非常适合候选人的职业发展需求，特别是在他刚刚建立独立实验室的关键时刻。在K08资助的早期阶段，候选人对癫痫的发育动物模型-Kv1.1钾通道基因敲除小鼠(即KcNA1缺失突变体)进行了详细的研究。后来，申请人继续研究生酮饮食(KD)抗惊厥疗效的潜在机制，这是一种公认但仍鲜为人知的治疗顽固性癫痫的方法。KD是一种高脂肪、低碳水化合物和低蛋白质的饮食，旨在模拟禁食后的早期生化变化。KD的特点是肝脏产生酮体。本研究的基本目的是评估GABA能系统在KD发作控制中的潜在作用，并在此过程中建立和验证KD的临床相关动物模型。作为第二个目标，将确定慢性酮体暴露是否可以增强大脑中的GABA能抑制。此外，KD是否可以在治疗期间以外发挥持久的抗癫痫作用的问题将被解决，从而为未来旨在确定KD抗癫痫作用的机制基础的研究奠定基础。为了达到这些目标，将研究KD对KcnA1缺失突变体的影响。此外，还将研究从这些小鼠制备的海马器型切片培养中酮小体的长期影响。拟议的研究将采用神经解剖学(即组织学、免疫细胞化学、原位杂交)和功能(如视频脑电监测、细胞电生理)技术。预计这些研究的结果将阐明KD在癫痫发生中的潜在作用，特别是与GABA能抑制作用有关的作用。

DESCRIPTION (provided by applicant): This application is for an Independent Scientist Award (K02). The candidate is a pediatric neurologist with a specialty interest in childhood epilepsy, and is currently in the final year of K08 (MCSDA) funding. He recently relocated to the University of California at Irvine (UCI). UCI hosts an internationally recognized basic neuroscience research community, and is well suited to the career development needs of the candidate, especially at this critical time when he has just established an independent laboratory. During the early period of K08 funding, the candidate performed detailed studies of a developmental animal model of epilepsy, the Kv1.1 potassium channel knockout mouse (i.e., the Kcna1-null mutant). Later, the applicant pursued research into the mechanisms underlying the anticonvulsant efficacy of the ketogenic diet (KD), an established but still poorly understood treatment for intractable epilepsy. The KD is a high-fat, low carbohydrate and low-protein diet designed to mimic the early biochemical changes seen upon fasting. The hallmark feature of the KD is the production of the ketone bodies by the liver. The fundamental goal of the proposed studies is to assess the potential role of the GABAergic system in contributing to seizure control by the KD, and in the process establish and validate a clinically relevant animal model of the KD. As a secondary goal, it will be determined whether chronic ketone body exposure can enhance GABAergic inhibition in the brain. Further, the question of whether the KD can exert a lasting antiepileptic effect, beyond the duration of therapy, will be addressed, thereby setting the stage for future studies aimed at determining a mechanistic basis for an anti-epileptogenic effect of the KD. Toward these goals, the effects of the KD on the Kcna1-null mutant will be investigated. In addition, the long-term impact of ketone bodies in hippocampal organotypic slice cultures prepared from these mice will be studied. Neuroanatomical (i.e., histological, immunocytochemical, in situ hybridization) and functional (i.e., video-EEG monitoring, cellular electrophysiological) techniques will be employed in the proposed studies. It is expected that the results of these investigations will shed light on the potential role of the KD in epileptogenesis, especially as it relates to effects on GABAergic inhibition