Cellular Mechanisms in Epileptogenesis

癫痫发生的细胞机制

基本信息

批准号：
7250931
负责人：
John R Huguenard
金额：
$ 34.15万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1978
资助国家：
美国
起止时间：
1978-01-01 至 2010-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7250931
关键词：
Ablation Absence Epilepsy Accounting Action Potentials Affect Affinity Agonist Aminobutyric Acid Aminobutyric Acids Animals Anticonvulsants Antiepileptic Agents Benzodiazepines Biological Assay Calcium Channel Cell Nucleus Cells Cessation of life Chromosome Pairing Chronic Clonazepam Data Defect Dose Elements Epilepsy Epileptogenesis Event Exhibits Fire - disasters Frequencies Generations Glutamates Goals Heterogeneity In Situ Hybridization In Vitro Injury Kinetics Lesion Maps Measurement Measures Mediating Membrane Methods Modeling Modification Molecular Mus Mutate Neurons Neurotransmitters Noise Output Pathway interactions Peptides Pharmacology Play Property Rattus Research Personnel Reverse Transcriptase Polymerase Chain Reaction Role Seizures Sensory Process Series Sleep Slice Specificity Structure Synapses System Techniques Testing Thalamic structure Translating Valproic Acid Vasoactive Intestinal Peptide base clinically relevant feeding gamma-Aminobutyric Acid in vivo inhibitory neuron man markov model neuropeptide Y nucleus reticularis patch clamp postsynaptic presynaptic programs receptor receptor function research study response synaptic inhibition uptake voltage voltage clamp

项目摘要

DESCRIPTION (provided by applicant): Experiments examine modulation of thalamic neuronal and network activities by the inhibitory neurotransmitter g-aminobutyric acid (GABA), and by neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP). One series of experiments focuses on molecular differences in GABA type A receptors in neurons of mouse nucleus reticularis (nRt) and the ventrobasal relay nucleus (VB), and the functional and pharmacological consequences of receptor subunit heterogeneity in these structures, whose reciprocal connectivity underlies thalamic rhythm generation. The hypothesis that alpha3 subunit-containing receptors in nRt cells confer sensitivity to the benzodiazepine anticonvulsant clonazepam will be tested in mutated mice. The role of beta3-containing receptors in generating prolonged inhibitory postsynaptic currents (IPSCs) that have an anti-rhythmogenic action in nRt will be examined. A second group of experiments explores actions of NPY and VIP on neuronal and circuit activities in mice and rats. The hypothesis that NPY is released during intense intrathalamic oscillatory activity, and in turn has anti-oscillatory effects, will be tested. The possibility that chronic dosing of the anticonvulsant valproic acid in vivo enhances expression of NPY in nRt cells, resulting in increased release, will be explored. VIP effects on membrane properties and synaptic currents in nRt and VB neurons, and on thalamic circuit oscillations will be studied. Techniques will include whole cell patch clamp recordings of IPSCs and voltage-dependent membrane currents; application of pharmacological agents; single cell RT-PCR from neurons of in vitro thalamic slices; in situ hybridization; the use of mice mutated for various alpha and beta GABAA receptor subunits; and measurements of peptides released from thalamic slices. The long-term goals are to understand the control of thalamic neuronal and circuit activities and potential abnormalities that may underlie pathophysiological states such as absence epilepsy.

描述（由申请人提供）：实验检查抑制性神经递质γ-氨基丁酸（GABA）、神经肽Y（NPY）和血管活性肠多肽（VIP）对丘脑神经元和网络活动的调节。一系列实验重点关注小鼠网状核 (nRt) 和腹基底中继核 (VB) 神经元中 GABA A 型受体的分子差异，以及这些结构中受体亚基异质性的功能和药理学后果，其相互连接是丘脑的基础节奏的产生。 nRt 细胞中含有 α3 亚基的受体赋予对苯二氮卓类抗惊厥药氯硝西泮敏感性的假设将在突变小鼠中进行测试。将检查含有 β3 的受体在产生延长的抑制性突触后电流 (IPSC) 中的作用，该电流在 nRt 中具有抗节律作用。第二组实验探讨了 NPY 和 VIP 对小鼠和大鼠神经元和回路活动的作用。 NPY 在强烈的丘脑内振荡活动期间释放，进而具有抗振荡作用的假设将得到检验。将探讨体内抗惊厥药丙戊酸的长期给药是否会增强 nRt 细胞中 NPY 的表达，从而导致释放增加。将研究 VIP 对 nRt 和 VB 神经元膜特性和突触电流以及丘脑回路振荡的影响。技术将包括 IPSC 的全细胞膜片钳记录和电压依赖性膜电流；药剂的应用；体外丘脑切片神经元的单细胞 RT-PCR；原位杂交；使用各种α和β GABAA受体亚基突变的小鼠；以及丘脑切片释放的肽的测量。长期目标是了解丘脑神经元和回路活动的控制以及可能构成失神癫痫等病理生理状态的潜在异常。