Sexual dimorphism in seizure control is KCC2 mediated

癫痫控制中的性别二态性是 KCC2 介导的

• 批准号: 6571785
• 负责人: Aristea S Galanopoulou
• 金额: $ 17.28万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6571785
• 关键词: GABA receptor; cAMP response element binding protein; calcium flux; cell parasexuality; cell sorting; chloride ion; confocal scanning microscopy; convulsions; developmental neurobiology; epilepsy; estradiol; gene expression; immunochemistry; in situ hybridization; laboratory rat; male; membrane transport proteins; microarray technology; muscimol; phosphorylation; potassium ion; protein structure function; stereotaxic techniques; substantia nigra; voltage /patch clamp

DESCRIPTION (provided by applicant): This application is for a Mentored Clinical Scientist Development Award (K08) for Dr. Aristea S. Galanopoulou, sponsored by Dr. Solomon L. Mosh. Dr Galanopoulou is an MD (Medical School of Athens) PhD (McGill University) trained in Neurology and Clinical Neurophysiology at Albert Einstein College of Medicine, where she is currently Assistant Professor. At McGill, Dr. Galanopoulou was trained in Molecular Biology and in vitro gene delivery methods. At Albert Einstein College of Medicine she has used in vivo experimental methods pertaining to seizure research. Dr. Galanopoulou's long- term goals are to employ novel molecular biology based approaches to develop effective treatments for epilepsy. The substantia nigra pars reticulata (SNR) has crucial role in seizure control. Intranigral infusions of muscimol, a GABAA receptor agonist, have sex, age and region specific effects on seizures. This proposal will study the role of the potassium-chloride co-transporter KCC2 in the sexual differentiation of GABAA responsive male infantile rat SNR neurons, which determine its function in seizure control. KCC2 switches hippocampal GABAA receptors from depolarizing to hyperpolarizing. Dr. Galanopoulou's findings suggest that the level of KCC2 in the SNR may determine whether GABAA receptor activation is pro-convulsant or not and correlate with sex-specific membrane responses and patterns of estradiol signaling. These data raise the possibility of accelerating the development of efficient control systems for seizures, by over expressing KCC2 and altering the sexual phenotype of male SNR. Dr. Galanopoulou will test whether in vivo over expression of KCC2 in the SNR of PN21 male rats abolishes: 1) the muscimol-induced depolarization and calcium rise, 2) the estradiol-mediated down regulation of phosphorylated CREB, 3) the muscimol-induced pro-convulsant SNR responses, and 4) will identify other genes regulated by KCC2, which suppress the testosterone-organized muscimol-sensitive proconvulsant effects of male rat SNR. This research will entail in vivo gene delivery using adeno-associated viruses, stereotactic surgery, immunochemistry, in situ hybridization, seizure induction, gramicidin perforated patch clamp, fura-2AM imaging, confocal microscopy, stereological cell counting, DNA microarrays and statistics. Albert Einstein College of Medicine will offer the necessary mentoring, laboratory space, equipment and training opportunities to achieve these goals.

描述（由申请人提供）：该申请是为Aristea S. Galanopoulou博士的指导临床科学家发展奖（K08），由Solomon L. Mosh博士赞助。 Galanopoulou博士是Albert Einstein医学院的神经病学和临床神经生理学培训的雅典医学院博士学位（麦吉尔大学），她目前是助理教授。在麦吉尔（McGill），Galanopoulou博士接受了分子生物学和体外基因递送方法的培训。在阿尔伯特·爱因斯坦医学院（Albert Einstein College），她在体内实验方法中使用了与癫痫发作研究有关的方法。 Galanopoulou博士的长期目标是采用新型的基于分子生物学的方法来开发有效的癫痫疗法。黑质nigra pars网状（SNR）在癫痫发作中具有至关重要的作用。 GABAA受体激动剂肌酚的腔内输注对癫痫发作具有性别，年龄和区域特异性影响。该提案将研究氯化钾共转运蛋白KCC2在GABAA响应性雄性婴儿大鼠SNR神经元的性分化中的作用，该雄性大鼠SNR神经元在癫痫发作控制中的功能。 KCC2将海马GABAA受体从去极化变为超极化。 Galanopoulou博士的发现表明，SNR中的KCC2水平可以确定GABAA受体激活是否促抗震颤，并与性别特异性的膜膜反应和雌二醇信号的模式相关。这些数据通过过度表达KCC2并改变男性SNR的性表型来加速癫痫发作的有效控制系统的发展。 Dr. Galanopoulou will test whether in vivo over expression of KCC2 in the SNR of PN21 male rats abolishes: 1) the muscimol-induced depolarization and calcium rise, 2) the estradiol-mediated down regulation of phosphorylated CREB, 3) the muscimol-induced pro-convul​​sant SNR responses, and 4) will identify other genes regulated by KCC2, which suppress the雄性大鼠SNR的睾丸激素组织敏感性的突出作用。这项研究将需要使用腺相关病毒，立体定向手术，免疫化学，原位杂交，癫痫诱导，gramicidin刺穿斑块夹，fura-2am成像，共焦显微镜检查，刻板细胞计数，DNA微射击和统计学。阿尔伯特·爱因斯坦医学院将提供必要的指导，实验室空间，设备和培训机会，以实现这些目标。