Connexins in Ischemia-Induced Neuronal Death

缺血引起的神经元死亡中的连接蛋白

• 批准号: 7154057
• 负责人: MICHAEL V L BENNETT
• 金额: $ 33.85万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-12-15 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7154057
• 关键词: 2' ,3' -Cyclic-Nucleotide Phosphodiesterases; Acute; Alzheimer' s Disease; Animals; Antisense Oligonucleotides; Astrocytes; Attenuated; Brain; Cardiac Surgery procedures; Cell Death; Cells; Cessation of life; Connexin 43; Connexins; Coupled; Coupling; Data; Development; Down-Regulation; Dyes; Epilepsy; Exhibits; Fire - disasters; Gap Junctions; Glial Fibrillary Acidic Protein; GluR2 subunit AMPA receptor; Glucose; HIV Encephalopathy; Hand; Heart Arrest; Hippocampus (Brain); Human; Image Analysis; Immunofluorescence Immunologic; In Situ Hybridization; Injury; Interneurons; Intervention; Ischemia; Isolectin; Laboratory Finding; Mediating; Metabolic; Methods; Modeling; Molecular; Mus; Myelin; Nerve Degeneration; Neurodegenerative Disorders; Neuroglia; Neurologic; Neuronal Injury; Neurons; Operative Surgical Procedures; Oxidative Stress; Oxygen; Parvalbumins; Patients; Pattern; Play; Property; Prosencephalon; Proteins; Pyramidal Cells; Rattus; Research; Resistance; Role; Slice; Thinking; Time; Trauma; Tubulin; Up-Regulation; Western Blotting; Wild Type Mouse; cell type; day; dentate gyrus; deprivation; extracellular; granule cell; hippocampal pyramidal neuron; immunocytochemistry; in vitro Model; in vivo; killings; neuronal survival; promoter; protein expression; research study; transmission process

Transient, but severe global ischemia, observed in patients during cardiac arrest and cardiac surgery or induced experimentally in animals, induces selective and delayed neurodegeneration. Pyramidal cells in CA1 are the most sensitive; CA3 and granule cells of the dentate gyrus (DG) are resistant to ischemic damage, and GABAergic interneurons in CA1 also survive. The molecular mechanisms underlying this pattern of neuronal death are not well understood. The proposed research aims to study the role of gap junctions during the several days of "maturation" of neuronal injury after global ischemia. Recent findings from this laboratory indicate that global ischemia triggers a selective upregulation of Cx36 (and Cx32) protein expression in GABAergic interneurons of the vulnerable CA1 at times prior to the onset of neuronal death, consistent with a role in the survival of these neurons. Moreover, CA1 neurons in Cx32 (Y/-) mice exhibit enhanced vulnerability to global ischemia-induced neuronal death. These data suggest that increased inhibition of pyramidal cells through synchronization of inhibitory interneurons may be neuroprotective. Gap junctions between astrocytes are also thought to have a role in postischemic neuronal death. Dying cells can kill resistant neighboring glial cells via glial "fratricide" (bystander death) and thereby propagate injury to neighboring regions. On the other hand, gap junctional coupling of astrocytes mediates metabolic cooperation among them and attenuates neuronal death in models of oxidative stress. The underlying hypothesis of this proposal is that gap junctions play important roles in determining neuronal death and survival following global ischemia. The research plan for the next five years focuses on changes in the abundance, distribution and molecular and biophysical properties of brain gap junctions following neurological insult. Specific Aims are 1. Characterize ischemia-induced alterations in connexin expression and gap junction properties in the vulnerable CA1 and resistant CA3 and dentate gyrus of rats and mice. Experiments will examine global ischemia-induced changes in coupling of inhibitory interneurons and expression of connexin proteins by immunocytochemistry and Western blotting and of connexin mRNAs by in situ hybridization and. Experiments will determine the effects of acute knockdown of specific connexins by antisense oligonucleotides on neuronal vulnerability and will examine neuronal vulnerability in Cx32(Y/-) mice, Cx36(-/-) mice and mice deficient in astrocyte Cx43. 2. Examine effects of oxygen/glucose deprivation on hippocampal slice cultures by immunocytochemistry, in situ hybridization and electrophysiological methods. To examine ischemia-induced changes in gap junction properties in acute slices and organotypic hippocampal slice cultures by electrophysiological methods and image analysis. The proposed research is expected to impact on the development of new treatment strategies for intervention in global ischemia, a debilitating and often fatal trauma associated with cardiac arrest in humans. Moreover, this study has important implications for research on other neurodegenerative disorders including focal ischemia, epilepsy, AIDS encephalopathy, and Alzheimer's disease.

在心脏骤停和心脏手术期间的患者中观察到短暂但严重的全脑缺血，或 在动物实验中诱导，诱导选择性和延迟的神经变性。锥体细胞 CA_1最敏感，CA_3和齿状回颗粒细胞对缺血性脑损伤有抵抗作用。 损伤后，CA 1中的GABA能中间神经元也存活。这背后的分子机制 神经元死亡的模式还不清楚。本研究旨在研究gap的作用 在全脑缺血后神经元损伤的“成熟”的几天期间，最近的调查结果 表明全脑缺血触发Cx 36（和Cx 32）蛋白的选择性上调 在神经元死亡开始之前的时间在脆弱的CA 1的GABA能中间神经元中的表达， 与这些神经元存活的作用相一致。此外，Cx 32（Y/-）小鼠中的CA 1神经元表现出 增强对全脑缺血诱导的神经元死亡的脆弱性。这些数据表明， 通过抑制性中间神经元的同步化来抑制锥体细胞可以是神经保护性的。间隙 星形胶质细胞之间的连接也被认为在缺血后神经元死亡中起作用。垂死的细胞可以 通过神经胶质“自相残杀”（旁观者死亡）杀死抗性邻近神经胶质细胞，从而将损伤传播到 邻近地区。另一方面，星形胶质细胞的缝隙连接偶联介导代谢 它们之间的合作，并减弱氧化应激模型中的神经元死亡。底层 该建议的假设是间隙连接在决定神经元死亡中起重要作用 和存活率之间的关系。未来五年的研究计划重点关注以下方面的变化： 脑间隙连接的丰度、分布以及分子和生物物理特性 神经损伤具体目标是1。表征缺血诱导的连接蛋白改变 在脆弱的CA 1和抵抗的CA 3和齿状回中的表达和缝隙连接特性 老鼠和老鼠。实验将检查全脑缺血诱导的抑制性神经元偶联的变化。 免疫细胞化学和蛋白质印迹法检测中间神经元和连接蛋白的表达， 通过原位杂交和.实验将确定急性敲除 特异性连接蛋白的反义寡核苷酸对神经元的脆弱性，并将检查神经元 Cx 32（Y/-）小鼠、Cx 36（-/-）小鼠和星形胶质细胞Cx43缺陷小鼠的易感性。2.检查影响 通过免疫细胞化学原位观察海马脑片培养物上的氧/葡萄糖剥夺 杂交和电生理学方法。检测缺血诱导的缝隙连接变化 通过电生理学方法测定急性切片和器官型海马切片培养物的性质， 图像分析这项拟议中的研究预计将影响新治疗方法的开发 全脑缺血的干预策略，这是一种与心脏病相关的使人衰弱且往往致命的创伤， 逮捕人类。此外，这项研究对其他神经退行性疾病的研究具有重要意义。 疾病，包括局灶性缺血、癫痫、AIDS脑病和阿尔茨海默病。

项目成果

SNAP-25 is a target of protein kinase C phosphorylation critical to NMDA receptor trafficking.

• DOI: 10.1523/jneurosci.4933-08.2010
• 发表时间: 2010-01-06
• 期刊: The Journal of neuroscience : the official journal of the Society for Neuroscience
• 作者: Lau CG;Takayasu Y;Rodenas-Ruano A;Paternain AV;Lerma J;Bennett MV;Zukin RS
• 通讯作者: Zukin RS

Single channel properties of pannexin-1 and connexin-43 hemichannels and P2X7 receptors in astrocytes cultured from rodent spinal cords.

• DOI: 10.1002/glia.24250
• 发表时间: 2022-12
• 期刊: GLIA
• 影响因子: 6.2
• 作者: Garre, Juan Mauricio;Bukauskas, Feliksas F.;Bennett, Michael V. L.
• 通讯作者: Bennett, Michael V. L.

An Acute Mouse Spinal Cord Slice Preparation for Studying Glial Activation ex vivo.

用于离体研究神经胶质激活的急性小鼠脊髓切片制备。

• DOI: 10.21769/bioprotoc.2102
• 发表时间: 2017
• 期刊: Bio-protocol
• 影响因子: 0.8
• 作者: Garré,JuanMauricio;Yang,Guang;Bukauskas,FeliksasF;Bennett,MichaelVL
• 通讯作者: Bennett,MichaelVL

Blockade of calcium-permeable AMPA receptors protects hippocampal neurons against global ischemia-induced death

• DOI: 10.1073/pnas.0505408102
• 发表时间: 2005-08-23
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Noh, KM;Yokota, H;Bennett, MVL
• 通讯作者: Bennett, MVL