Connexins in Neuronal and Glial Gap Junctions in the Central Nervous System

中枢神经系统神经元和胶质间隙连接中的连接蛋白

• 批准号: 7849506
• 负责人: JOHN E RASH
• 金额: $ 38.94万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7849506
• 关键词: Area; Arousal; Autistic Disorder; Binding; Brain; Brain region; Caliber; Cells; Cerebral cortex; Chemicals; Cognition; Cognitive; Communication; Complex; Confocal Microscopy; Connexins; Connexon; Conscious; Coupled; Coupling; Data; Data Correlations; Detection; Diffusion; Discrimination; Disease; Electrical Synapse; Electron Microscopy; Epilepsy; Event; Freeze Fracturing; Frequencies; Functional disorder; Gap Junctions; General Anesthesia; Glutamate Receptor; Health; Hippocampus (Brain); Human; Immunofluorescence Microscopy; In Situ Hybridization; Individual; Intercellular Junctions; Interneurons; Ion Channel; Ion Exchange; Ions; Label; Lasers; Learning; Link; Long-Term Potentiation; Maps; Measurement; Measures; Memory; Messenger RNA; Metabolic; Methods; Microscopic; Microtomy; Molecular; Mus; N-Methylaspartate; Nature; Neuraxis; Neurons; Neurotransmitter Receptor; Neurotransmitters; Parkinson Disease; Pharmaceutical Preparations; Phenotype; Prevalence; Proteins; Regulation; Relative (related person); Reporting; Retina; Rodent; Role; Scanning; Schizophrenia; Second Messenger Systems; Site; Sleep; Spinal Cord; Structure; Synapses; Thinking; Time; Tracer; awake; base; cell type; connexin 36; design; gene discovery; gray matter; human disease; immunocytochemistry; motor control; public health relevance; second messenger; virtual

DESCRIPTION (provided by applicant): Gap junctions are microscopic intercellular junctions that provide for direct intercellular ionic, electrical, and metabolic coupling between nerve cells in the brain, retina, and spinal cord. Previously, gap junctions between neurons were thought to consist of hundreds or thousands of connecting channels ("connexons"), but they were also thought to be rare, to occur only between a few types of neurons, and to occur only in limited, non- cognitive areas of the central nervous system. The discovery of "miniature" gap junctions (<100 connexons) and preliminary evidence for their abundance throughout the brain suggests that "mini" gap junctions, particularly those at "mixed" (chemical plus electrical) synapses, may provide the structural basis for the detection of tiny sub-threshold electrical "spikelets" or "partial spikes" in principal neurons that are distributed throughout the brain. Gap junctions / partial spikes are thought to be essential for regulating and optimizing the high-frequency neuronal oscillatory synchronizations that are thought to underlie consciousness, arousal from sleep, cognition, associative binding for learning and memory, and fine motor control, and which become pathologically altered in epilepsy, schizophrenia, Parkinson's disease, and autism spectral disorders. We will combine laser scanning confocal immunofluorescence microscopy with newly-developed freeze-fracture replica immunogold labeling (FRIL) electron microscopy to detect, quantify, determine the protein composition of, and measure the sizes of gap junctions throughout the complex circuitry of the mouse brain, to make detailed measurements of "mini" gap junctions in cerebral cortex and hippocampus of both mouse and human brain, and to identify the neuronal subtypes linked by "mini" gap junctions. We will emphasize analysis of those regions that are primarily responsible for thinking and consciousness (cerebral cortex) and for learning and memory (hippocampus) and that represent the primary sites of origin of epileptic discharges. These complementary approaches will allow direct correlation of data from large-scale (whole mouse brain) to ultrastructural and molecular levels. The data to be obtained will be essential for understanding how consciousness is created, what distinguishes sleeping vs. awake states, how consciousness is altered during general anesthesia, and how memories are created, and will also be essential for identifying subcellular sites that may become targets for designing new drugs to treat disorders of electrical synaptic communication in the brain. PUBLIC HEALTH RELEVANCE A newly-discovered class of "miniature" gap junctions, found to be abundant between many classes of neurons, may provide the structural basis for propagation of small electrical "spikelets" between neurons, and thereby, to regulate the oscillatory synchronizations of cerebral cortex and hippocampus that are associated with human consciousness and for associative binding for learning and memory. We will detect, map, quantify, and determine the connexin protein composition of "mini" gap junctions throughout mouse brain, both at electrical and at "mixed" (chemical plus electrical) synapses, and compare the distributions of gap junctions and their constituent connexins proteins in mouse vs. normal human cerebral cortex and hippocampus. These data will serve as a prelude to measuring changes in these structures in human disease such as epilepsy, schizophrenia, Parkinson's disease, and autism spectral disorders.

描述（由申请人提供）：间隙连接是微观的细胞间连接，可提供直接的细胞间离子，电和代谢耦合，并在大脑，视网膜和脊髓中的神经细胞之间进行。以前，神经元之间的间隙连接被认为由数百或数千个连接通道（“连接”）组成，但是它们也被认为很少见，仅在几种类型的神经元之间发生，并且仅发生在中枢神经系统的有限的非认知区域。发现“微型”差距连接（<100个连接子）和初步证据，证明它们在整个大脑中的丰度都表明，“迷你”差距连接处，尤其是那些在“混合”（化学加上电气）突触的差距，可能为在整个脑中的微小电气“ spikelels”中的微小spikeleons进行了分布，可以为检测微小的spikeles spikeles neurons散布提供结构性基础。 Gap junctions / partial spikes are thought to be essential for regulating and optimizing the high-frequency neuronal oscillatory synchronizations that are thought to underlie consciousness, arousal from sleep, cognition, associative binding for learning and memory, and fine motor control, and which become pathologically altered in epilepsy, schizophrenia, Parkinson's disease, and autism spectral disorders. We will combine laser scanning confocal immunofluorescence microscopy with newly-developed freeze-fracture replica immunogold labeling (FRIL) electron microscopy to detect, quantify, determine the protein composition of, and measure the sizes of gap junctions throughout the complex circuitry of the mouse brain, to make detailed measurements of "mini" gap junctions in cerebral cortex and小鼠和人脑的海马，并确定由“迷你”间隙连接连接的神经元亚型。我们将强调对主要负责思维和意识（脑皮质）和学习和记忆（海马）的区域的分析，并代表癫痫放电起源的主要部位。这些互补方法将允许将大规模（整个小鼠大脑）与超微结构和分子水平的数据直接相关。要获得的数据对于理解如何创造意识，与睡眠状态，清醒状态的区别，意识如何改变以及如何创造记忆是至关重要的，这对于确定可能成为设计新药物以治疗大脑中电气突触传播的新药物的目标的细胞网站也是必不可少的。公共卫生相关性在许多类别的神经元之间发现丰富的新发现的“微型”差距连接可能会为在神经元之间传播小电气“ Spikelets”的结构基础，从而使振动性与人类意识与人类意识相关，从而调节振动性的振动性同步。我们将检测，绘制，量化，确定和确定整个小鼠大脑中“ mini”间隙连接的连接蛋白组成，无论是在电气和“混合”（化学加上电气）突触，并比较间隙连接点及其组成蛋白的分布及其组成蛋白的分布。这些数据将成为衡量人类疾病中这些结构变化的序幕，例如癫痫，精神分裂症，帕金森氏病和自闭症谱系障碍。