Functional Study of Inhibitory and Excitatory Neurotransmission in the Nervous Tissue Resected From Human Brain: Understanding New Molecular Mechanisms and Discovering New Therapeutic Targets to Cure Drug-resistant Epilepsy.
试验速览
- 阶段
- 不适用
- 状态
- 招募中
- 发起方
- 入组人数
- 60
- 试验地点
- 1
- 主要终点
- Composition of GABA-A receptors that modulate macroscopic currents
研究概览
简要总结
Epilepsy is a neurological condition that afflicts 1% of the world population. 30% of patients become drug-resistant to classic antiepileptic treatment and only a small percentage, 5%, can undergo a neurosurgical resection of epileptic focus and recover almost completely from symptoms. To date, an imbalance between inhibitory and excitatory neurotransmission has been well accepted as the main root cause of epilepsy. A better understanding of the molecular mechanisms of this can lead to developing new therapeutic strategies. The investigators of the project want to describe the functional alteration of GABA- A receptor, the main actor of inhibitory neurotransmission in the central nervous system and characterize its subunit composition in the epileptic foci of patients with temporal lobe epilepsy. The authors, also, want to modulate, by means of selective neuroactive molecules, the function of this receptor to increase the inhibitory tone in the epileptic brain.
详细描述
The project presents 3 aims:
- Characterization of macroscopic GABA-A mediated currents evoked by the specific agonist, muscimol, in presence of selective negative and positive allosteric modulators to identify the subunit composition.
- Functional characterization of synaptic GABA-A mediated currents recorded from hippocampal and cortical pyramidal neurons by using pharmacological tools.
- After the acquired knowledge of aims 1) and 2), promotion of the increase of inhibitory tone in epileptic tissue, enhancing the GABA release from interneurons onto principal cells, by stimulation of other neurotransmitter systems present in the brain: cholinergic system, serotonergic system, and dopaminergic system.
Experimental design:
AIM1 WP1.1: Patch-clamp recordings of whole-cell GABA-A mediated currents from pyramidal and interneurons evoked by muscimol, perfusing the slices with positive allosteric modulators such as diazepam or nitrazepam, that are selective for apha1/alpha2 subunits GABA-A containing receptors or phenobarbital selective for alpha4/alpha6 subunits GABA-A containing receptors.
WP 1.2: Patch-clamp recordings of whole-cell GABA-A mediated currents from pyramidal and interneurons evoked by muscimol, perfusing the slices with negative allosteric modulators such as BetaCCT, which is selective for apha1/alpha2 subunits GABA-A containing receptors or L655,708 and Furosemide that are selective for alpha4/alpha6 subunits GABA-A containing receptors.
研究设计
- 研究类型
- Observational
- 观察模型
- Case Only
- 时间视角
- Retrospective
入排标准
- 性别
- All
- 接受健康志愿者
- 否
入选标准
- •Patients with drug-resistant temporal lobe epilepsy.
排除标准
- •Patients suffering from Temporal lobe epilepsy caused by stroke, ischemia, cavernous angiomas, or meningiomas growth.
结局指标
主要结局
Composition of GABA-A receptors that modulate macroscopic currents
时间窗: 24 months
WP1.1- Slices of tissue obtained from 12 patients Tratment :diazepam or nitrazepam and phenobarbital WP 1.2:- Slices of tissue obtained from 12 patients Treatment: BetaCCT and L655,708 or Furosemide .
Modulation of synaptic transmission by GABA-A receptors PAMs and NAMs
时间窗: 24
WP2.1 Slices of tissue obtained from 12 patients Treatment: diazepam, phenobarbital, BetcCCt, and Furosemide, effect on inhibitory synaptic transmission WP2.2: Slices of tissue obtained from 12 patients Treatment: diazepam, phenobarbital, BetcCCt, and Furosemide, effect on excitatory synaptic transmission
Modulation of inhibitory synaptic transmission by cholinergic, dopaminergic and serotonergic PAMs.
时间窗: 12
WP 3.1 Slices of tissue obtained from 12 patients Based on results obtained in Primary Outcome 1 and 2 Treatment: PNU dFBr, mCPGB, or L-dopa, effect on inhibitory neurotransmission.
次要结局
未报告次要终点
研究者
Katiuscia Martinello
Researcher, PhD
Neuromed IRCCS