Isolation of Ictogenic Cells and Circuits Within the Parahippocampal Region

海马旁区域内致炎细胞和回路的分离

• 批准号: 9270622
• 负责人: Sanjay S Kumar
• 金额: $ 31.99万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9270622
• 关键词: Acute; Address; Adult; Anatomy; Animal Model; Animals; Anticonvulsants; Attention; Biological Assay; Brain; Categories; Cells; Chronic; Coupling; Diagnosis; Disinhibition; Electrophysiology (science); Epilepsy; Epileptogenesis; Excitatory Synapse; Focal Seizure; Functional disorder; Glutamates; Goals; Hippocampus (Brain); Histologic; Human; Interneurons; Intervention; Knowledge; Laboratories; Lasers; Lateral; Location; Measures; Medial; Mediating; Methods; Microfluidic Microchips; Modeling; Modernization; Morphology; Neurons; Pathology; Pharmacology; Physiological; Pilocarpine; Population; Prevention; Property; Replacement Therapy; Research; Role; Sampling; Scanning; Slice; Source; Structure; Synapses; Techniques; Temporal Lobe; Temporal Lobe Epilepsy; Testing; Therapeutic Intervention; Time; Tissues; base; biocytin; cell type; combat; design; entorhinal cortex; excitatory neuron; hippocampal pyramidal neuron; inhibitory neuron; insight; neuronal excitability; novel; optogenetics; response; spatiotemporal; tool

Project Summary / Abstract Temporal lobe epilepsy (TLE) is the most common form of human epilepsy, often intractable to anticonvulsant therapy, whose treatment and prevention are critically dependent on understanding pathophysiological mecha- nisms in interlinked structures of the temporal lobe. The long-term objective of the proposed research is to iden- tify and isolate components of the underlying epileptogenic circuits within two of these structures and to decipher mechanisms responsible for rendering them epileptic. The presubiculum (PrS) and parasubiculum (Par), rela- tively obscure structures lying at the interface between the hippocampus and entorhinal cortex (ERC), have been anatomically implicated in rendering neurons in the media entorhinal area (MEA) hyperexcitable during TLE. The proposed research is aimed at characterizing the PrS/Par physiologically and determining which of the recently identified neuronal populations in PrS/Par hyperexcitable become during TLE (Specific Aims 1 & 2). Anatomical studies have suggested that GABAergic projection neurons in PrS are functionally connected with inhibitory interneurons in LIII of MEA raising the possibility of PrS/Par-mediated control of local inhibitory circuits in MEA through disinhibition. However, neither inhibitory neurons of PrS/Par nor their projections to the MEA have been physiologically characterized. The proposed research will test this hypothesis by photouncaging glutamate fo- cally to activate GABAergic neurons in PrS/Par while recording inhibitory synaptic responses from interneurons in MEA. Additionally, the alternative hypothesis that PrS is the source of persistent excitatory synapses to both excitatory and inhibitory neurons in LIII that perish during TLE will also be tested (Specific Aim 3). Without a systematic and deliberate effort to address these issues, it will be difficult to make any real progress in tracing the origins of TLE, determining its progress, and identifying what steps, if any, can be taken to intervene (identify location and means) and even to halt its progress. To this end electrophysiological, pharmacological and neuro- anatomical techniques would be used in conjunction with a well-established animal model of TLE to characterize the PrS physiologically and test key hypotheses outlined above. Laser-scanning photouncaging of glutamate will be combined with CESOP and used in lieu of the labor-intensive paired recordings for a rapid, near-complete as- sessment of functional synaptic connectivity between PrS/Par and MEA as well as to measure the extent of synaptic reorganization that occurs in these structures during TLE.

项目摘要/摘要 颞叶癫痫(TLE)是人类最常见的癫痫类型，通常对抗惊厥药物难以治愈 治疗，其治疗和预防关键依赖于对其病理生理机制的了解。 在颞叶的相互关联的结构中的神经。拟议研究的长期目标是认识到- 识别并分离其中两个结构内潜在致痫回路的组件，并破译 使他们成为癫痫患者的机制。下丘前(Prs)和下丘旁(PAR)，相对。 位于海马体和内嗅觉皮质(ERC)交界处的隐蔽结构，已经被 解剖学上涉及在TLE期间使内嗅区(MEA)中的神经元过度兴奋。这个 拟议的研究目的是描述PRS/PAR的生理特征，并确定最近 在TLE期间，在PRS/PAR中已识别的神经元群体变得过度兴奋(特定目标1和2)。解剖学 研究表明，PR内的GABA能投射神经元在功能上与抑制性有关 大脑中动脉LIII内的中间神经元增加了PRS/PAR介导的大脑中动脉局部抑制回路的控制 通过解除抑制。然而，无论是PrS/PAR抑制性神经元还是它们向MEA的投射都没有 具有生理特征的。这项拟议的研究将通过对谷氨酸进行光老化来验证这一假设。 在记录中间神经元抑制性突触反应的同时，CALY激活PrS/PAR中的GABA能神经元 在中东和非洲地区。此外，另一种假设认为，中枢神经系统是持久兴奋性突触的来源 在TLE期间死亡的LIII中的兴奋性和抑制性神经元也将被测试(特定目标3)。如果没有 如果不作出系统和刻意的努力来解决这些问题，就很难在追查方面取得任何实际进展 TLE的起源，确定其进展，并确定可以采取哪些步骤进行干预(确定 地点和手段)，甚至停止其进展。为此，电生理、药理学和神经学- 解剖学技术将与已建立的TLE动物模型结合使用，以表征 从生理和测试两个方面对上述关键假设进行了研究。谷氨酸遗嘱的激光扫描光老化 与CESOP相结合，用于代替劳动密集型配对录音，实现快速、接近完成的AS- PRs/PAR与MEA之间功能性突触连接的评估及突触范围的测量 在TLE期间在这些结构中发生的重组。