BRAIN GALANIN RECEPTORS AND EPILEPSY

描述（由申请人提供）：该项目的长期目标是了解神经肽丙氨酸抑制癫痫发作活动的机制。我们的实验室首先证明了癫痫发作会改变海马体中的丙氨酸，并且丙氨酸在癫痫持续状态中具有深远的癫痫保护作用。该项目有三个目标。首先，我们将开发一种新的方法来下调幼稚动物的丙氨酸受体，以研究丙氨酸受体在癫痫发作中的作用。这将通过开发和优化针对丙氨酸受体1和2亚型的肽核酸（PNA）反义来实现。PNA对甘丙氨酸受体的影响将通过研究甘丙氨酸受体结合、免疫细胞化学和Western blot在甘丙氨酸受体转染的细胞系和原代神经元培养中进行研究。其次，我们将测试大脑中甘丙肽受体亚型的下调选择性地增加动物癫痫发作的易感性，并增加癫痫相关的神经元损伤的假设。我们将研究未经处理和甘丙氨酸受体pna处理的大鼠在自我维持癫痫持续状态期间癫痫发作活动的启动和维持方面的差异，并将比较对照组和pna处理的大鼠的神经元损伤程度。第三，我们将研究丙氨酸受体刺激对癫痫保护作用的推测机制。具体来说，我们将研究甘丙氨酸受体PNA治疗如何影响第二信使系统（环磷酸腺苷、磷酸二肽周转、丝裂原激活的蛋白激酶活性、兴奋性氨基酸谷氨酸释放），并将这些影响与PNA对癫痫状态的影响联系起来。该项目将为了解参与内源性癫痫发作抑制的大脑机制提供重要数据，并将证明丙氨酸受体活性配体作为治疗癫痫的新方法是合理的。

DESCRIPTION (provided by applicant): The long-term objective of this project is to understand the mechanisms involved in inhibition of seizure activity by the neuropeptide galanin. Our lab was the first to show that seizures alter galanin in the hippocampus and that galanin has a profound seizure-protective effect in status epilepticus. The project has three goals. First, we will develop a novel approach to down-regulate galanin receptors in naive animals, in order to study the role of galanin receptors in seizures. This will be achieved by developing and optimizing peptide nucleic acid (PNA) antisense targeted against galanin receptor subtypes 1 and 2. The effects of PNA on galanin receptors will be examined by studying galanin receptor binding, immunocytochemistry, and Western blot in galanin receptor-transfected cell lines and primary neuronal cultures. Second we will test the hypothesis that down-regulation of galanin receptor subtypes in the brain selectively increases animals' predisposition to seizures, and increases seizure-related neuronal injury. We will examine the differences between naive and galanin receptor PNA-treated rats in the initiation and maintenance of seizure activity during self-sustaining status epilepticus, and will compare the degree of neuronal injury between control and PNA-treated rats. Third, we will study putative mechanisms that underlie seizure-protective effects of galanin receptor stimulation. Specifically, we will study how galanin receptor PNA treatment affects second messenger systems (cyclic adenosine monophosphate, phosphoinisitide turnover, mitogen-activated protein kinase activity, excitatory amino acid glutamate release) and will correlate these effects with the effects of PNA on status epilepicus. The proposed project will yield important data for understanding brain mechanisms that participate in endogenous suppression of seizures, and will justify the use of active ligands of galanin receptors as a new approach for the treatment of epilepsy.