Effects of pregabalin and thrombospondins on enhanced excitatory connectivity, new synapse formation and epileptogenesis after neocortical injury

普瑞巴林和血小板反应蛋白对新皮质损伤后兴奋性连接增强、新突触形成和癫痫发生的影响

• 批准号: 8802778
• 负责人: David Allan Prince
• 金额: $ 37.07万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8802778
• 关键词: Affect; Aftercare; Anesthesia procedures; Animal Model; Animals; Astrocytes; Axon; Binding; Brain; Brain Injuries; Cerebral cortex; DLG4 gene; Development; Elvax; Epilepsy; Epileptogenesis; Excitatory Synapse; Frequencies; Future; Glial Fibrillary Acidic Protein; Glutamates; Health; Hippocampus (Brain); Human; In Vitro; Incidence; Injury; Knockout Mice; Lasers; Lead; Lidocaine; Life; Link; Maps; Measures; Modeling; Mus; Neocortex; Neuroglia; Neurons; Pharmaceutical Preparations; Process; Prophylactic treatment; Pyramidal Cells; Recovery; Role; Scanning; Seizures; Slice; Spinal nerve structure; Status Epilepticus; Stroke; Structure; Synapses; Techniques; Testing; Thrombospondin 1; Thrombospondins; Time; Trauma; Traumatic Brain Injury; Whole-Cell Recordings; afferent nerve; allodynia; axonal sprouting; density; dorsal horn; gabapentin; hippocampal pyramidal neuron; in vivo; injured; knockout animal; neocortical; nerve injury; novel strategies; overexpression; pregabalin; prevent; prophylactic; receptor; research study; synaptogenesis; treatment duration

DESCRIPTION (provided by applicant): The effects of gabapentin and thrombospondins on enhanced excitatory connectivity, new synapse formation and epileptogenesis after neocortical injury sprouting of new excitatory wiring between neurons in cerebral cortex and hippocampus, changes in glial cells and formation of excitatory synapses are consequences of brain injury that are thought to contribute to epilepsy in animal models and human brain. There is no effective prophylactic treatment available to prevent epileptogenesis after brain injury. The planned experiments focus on a new approach for limiting the development of the increased excitatory connections and epilepsy after cortical trauma. Astrocyte-secreted thrombospondins (TSPs) are involved in new excitatory synapse formation during development and after cortical injury. Experiments will test the hypothesis that pregabalin (PGB) (Lyrica), an approved drug that interferes with the binding of TSPs to their alpha2delta-1 receptor, will decrease excitatory synapse formation and sprouting and limit development of epileptiform activity. The effects of PGB and TSPs will be explored in the partial cortical isolation ("undercut", UC) model of epileptogenic neocortical injury in naïve mice, in TSP "knockout" mice and alpha2delta-1 overexpressing epileptic mice. The incidence of epileptiform activity recorded in in vitro cortical slices after injury, sprouting of axons, density of excitatory synapses and network connectivity will be measured using electrophysiological and anatomical techniques. Laser scanning photostimulation of caged glutamate will be used in conjunction with whole cell recordings to map excitatory connections in cortical slices. A possible link between excessive neuronal activity and increases in TSPs, the alpha2delta-1 receptor and new synapse formation will be studied in naïve or injured cortex. The effects of PGB treatment after cortical injury in vivo on these measures will be assessed to test the hypothesis that the drug will decrease the structural and functional abnormalities that follow brain trauma and lead to the development of epilepsy. Relevance: Posttraumatic epilepsy is a prominent consequence of serious neocortical or hippocampal injury that alters neuronal and glial structure and function and induces hyperexcitability in cortical circuits. Unfortunately, treatment is often ineffective at relieving seizures once they occur and no drug is available that will prevent the epileptogenic processes that lead to posttraumatic epilepsy. Results of these experiments will contribute to understanding cellular and circuit effects of cortical injury, and provide new information about a potential role for gabapentin and pregabalin to prevent development of epilepsy after brain injury.

描述（由申请人提供）：加巴喷丁和血小板反应蛋白对新皮质损伤后兴奋性连接增强、新突触形成和癫痫发生的影响、大脑皮层和海马神经元之间新兴奋性线路的萌发、神经胶质细胞的变化和兴奋性突触形成的后果动物模型和人类大脑中被认为导致癫痫的脑损伤目前尚无有效的预防治疗方法。脑损伤后的癫痫发生。计划的实验重点是限制皮质创伤后兴奋性连接增加和癫痫的发展，星形胶质细胞分泌的血小板反应蛋白（TSP）参与发育过程中和皮质损伤后新的兴奋性突触的形成。将测试普瑞巴林 (PGB) (Lyrica) 的假设，普瑞巴林是一种已批准的药物，可干扰 TSP 与其 alpha2delta-1 受体的结合，从而减少兴奋性突触形成和发芽以及限制癫痫样活动的发展将在幼稚小鼠、TSP“敲除”小鼠和 alpha2delta 致癫痫性新皮质损伤的部分皮质隔离（“undercut”，UC）模型中进行探讨。 -1 过表达癫痫小鼠的体外皮质记录的癫痫样活动的发生率。 损伤后的切片、轴突的萌芽、兴奋性突触的密度和网络连接将使用电生理学和解剖学技术来测量，笼状谷氨酸的激光扫描光刺激将与全细胞记录结合使用，以绘制皮质切片中的兴奋性连接。在过度的神经元活动和 TSP 增加之间，将在幼稚或受损的皮层中研究 alpha2delta-1 受体和新突触的形成。将评估皮质损伤后体内治疗的这些措施，以检验该药物将减少脑损伤后的结构和功能异常并导致癫痫发展的假设。海马损伤会改变神经元和神经胶质的结构和功能，并引起皮质回路过度兴奋，不幸的是，一旦癫痫发作，治疗通常无法有效缓解，并且没有药物可以预防癫痫发作。这些实验的结果将有助于了解皮质损伤的细胞和回路效应，并提供有关加巴喷丁和普瑞巴林预防脑损伤后癫痫发展的潜在作用的新信息。