Molecular Mechanisms Of Synaptic Transmission

突触传递的分子机制

• 批准号: 8349615
• 负责人: David Robinson
• 金额: $ 148.78万
• 依托单位: NATIONAL INSTITUTE ON DEAFNESS AND OTHER COMMUNICATION DISORDERS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8349615
• 关键词: Addendum; Adhesions; Adult; Affect; Alzheimer' s Disease; Amygdaloid structure; Anesthesiology; Auditory; Autistic Disorder; Axon; Bacteria; Baltimore; Biochemistry; Biology; Brain; Cell Adhesion Molecules; Cell Culture Techniques; Cells; Cessation of life; Child; Chronic Obstructive Airway Disease; Cochlea; Collaborations; Communication; Core Facility; Dendrites; Development; Disease; Electron Microscope; Equipment; Erinaceidae; Excitatory Synapse; Farming environment; Future; GABA Receptor; Glutamate Receptor; Growth Cones; Hair Cells; Health Sciences; Institutes; Investigation; Journals; Kinocilium; Knock-in Mouse; Laboratories; Labyrinth; Lasers; Lead; Location; Maintenance; Manuscripts; Membrane Proteins; Methods; Microscope; Microtomy; Molecular; Mus; Mutation; N-Methyl-D-Aspartate Receptors; N-Methylaspartate; National Heart, Lung, and Blood Institute; National Institute of Child Health and Human Development; National Institute of Mental Health; National Institute of Neurological Disorders and Stroke; National Institute on Deafness and Other Communication Disorders; Nervous system structure; Neuraxis; Neurology; Neuronal Plasticity; Neurons; Neurosciences; Otitis Media; Pain; Paper; Pathology; Physiology; Potassium Channel; Preparation; Process; Proteins; Publishing; Rattus; Regulation; Research; Research Personnel; Research Project Grants; Respiratory Tract Infections; Retina; Schizophrenia; Signal Transduction; Spinal Cord; Supporting Cell; Synapses; Synaptic Cleft; Synaptic Transmission; System; Taro Vegetable; Techniques; Training; Ukraine; United States National Institutes of Health; Vaccines; Vertebral column; Work; Writing; comparative; deafness; dorsal cochlear nucleus; human SMO protein; interest; kainate; medical schools; meetings; member; mouse model; mutant; nervous system disorder; neuronal growth; neuropathology; posters; presenilin; protein transport; receptor; scaffold; synaptic function; trafficking; transmission process

Since the death of Robert Wenthold in October, 2009, the lab has continued to support the projects of some of the researchers who had moved to the lab of Dr. Katherine Roche in NINDS (Elizabeth Webber, Kai Chang, Takaaki Hirai, as well as Chan-Ying Zheng who transferred this year from the lab of Dr. Kachar), as well as Gail Seabold, who has continued to work mainly in the lab here, although her PI is Sharon Milgram from NHLBI. The lab has continued to do some neuron cell culture preparations as needed by these researchers. We also oversee maintenance of the JEOL 1010 transmission electron microscope and related equipment, as well as the Zeiss LSM710 laser confocal microscope and Leica Biowave System, which continue to be used here by some of these researchers. In addition, reviews of manuscripts, both of other researchers at NIDCD, and reviews requested by various journals, are performed regularly. Basically, work now is done as collaborations with PIs of other labs, and we will become a separate core facility in October, the beginning of the 2012 fiscal year. Our main obligation will be to facilitate the projects of other PIs in the NIDCD. Some of the collaborations here continue the work done in the lab of Dr. Wenthold on the trafficking of glutamate receptors and associated proteins of the neuronal synapse. However, as part of our commitment to helping NIDCD studies, the lab has become involved more and more in studies of development and pathology of the cochlea, and in the near future the lab plans to develop some work on central auditory function. The specific research projects that the lab has been involved in as collaborators this year are as follows: NIDCD A With researchers of the original Wenthold/NIDCD lab: 1) Completion of a study with Philip Wang and Stephan Brenowitz on NMDA receptors on neuronal growth cones, recently submitted. Performed EM analyses used in this study and also provided the cultured neurons. Now published in the Journal of Neuroscience. 2) Continued work on a SALM1 paper with Gail Seabold and Philip Wang. This is an adhesion protein associated with glutamate receptors at synapses. Performed EM analyses used in this study and provided the cultured neurons. This paper is in revision following review by the Journal of Biological Chemistry. (posters presented at meetings also) 3) Worked with Chan-Ying Zheng to complete a video method paper on FRAP of synaptic spines now published in the journal JoVE. 4) Worked with Chan-Ying Zheng to complete a paper as an addendum to the published paper on PSD-95 and SAP102. This addendum is now published in the journal Communicative & Integrative Biology. 5) The lead investigator was invited to write a review paper on MAGUK plasticity for The Neuroscientist, and enlisted the help of Chan-Ying Zheng, Gail Seabold and Martin Horak. This now is in press and is published on-line. NIDCD B With others at NIDCD: 1) Worked with Dabin Ren from the lab of Dr. Xin-Xing Gu to complete 2 separate EM studies of immunogold localization on bacteria associated with Otitis Media in children and respiratory infections, including COPD, in adults. One has now been published in the journal Vaccine. A second paper is in review. 2) Worked with researchers from the lab of Dr. Andy Griffith on a study of changes in the cochlea involving mutants associated with deafness. This paper is now in press in J. Clin. Inv. (posters presented at meetings also) 3) Trained and working with Kate Szarama from the labs of Drs. Matt Kelley and Richard Chadwick looking at the ultrastructural changes in the cochlea related to development and disease. One paper is in review and another will be submitted soon. (posters presented at meetings also) 4) Working with Helen May-Simera from the lab of Dr. Matt Kelley (in collaboration with Kate Szarama) on immunogold localization of kinocilium proteins in the developing cochlea. 5) Completed, as noted above, 3 projects/papers with Chan-Ying Zheng while she was associated with the lab of Dr. Kachar. 6) Involved in some preliminary studies of EM immunogold distribution with researchers in the lab of Dr. Tom Friedman. Especially involved in training and working with Inna Bellyantseva on EM techniques. 7) Working on collaboration with members of the lab of Dr. Paul Sieving on EM localization of a surface protein in the retina of experimental mice. 8) Providing thin sections for Dr. Bechara Kachar in ultrastructural studies of hair cells and the cochlea. 9) Advising Janaina Brusco from the lab of Dr. Bechara Kachar on an EM immunogold and 3D study of synapses of the amygdala. 10) Advising Taro Fujikawa from the lab of Dr. Bechara Kachar on a study of kainate glutamate receptors in the cochlea. 11) Planning immunoEM studies on the dorsal cochlear nucleus for the lab of Dr. Stephan Brenowitz. 12) Planning EM studies of inner ear with Dr. Doris Wus lab. With other NIH institutes: 6) Working on several studies of EM localization of sonic hedgehog and its receptors, patched and smoothened, with Drs. Pamela Yao and Mark Mattson of NIA. A first author paper now is in press in the Journal of Comparative Neurology. We also have another first author manuscript in review in the Journal of Comparative Neurology, and an addendum to the former paper and other studies are in preparation. (posters presented at meetings also) 1) Involved in 2 preliminary studies on mutations of proteins that affect the nervous system (including a mouse model for schizophrenia) with Drs. Jie-Min Jia and Song Jiao of NIMH (manuscript in preparation). 2) Worked with the lab of Dax Hoffman in NICHD on a potassium channel auxiliary subunit involved in synaptic integration and plasticity. Paper in press in the journal Neuron. With labs outside of NIH: 1) Worked with Dr. Jinhyun Kim of Janelia Farm Research Campus (HHMI), on EM immunolocalization of a protein in the synaptic cleft. A manuscript is in review. 2) Worked on EM immunogold contributions to 3 papers related to pain effects on neuronal plasticity of the spinal cord with Dr. Yuan-Xiang Tao of Johns Hopkins School of Medicine in Baltimore; these involve other collaborators such as Drs. Richard Huganir and Jeffrey Rothstein of Johns Hopkins, and D. Olga Kopach of the Bogomoletz Institute of Physiology in the Ukraine. All 3 papers are now published in the journals Anesthesiology, Molecular Pain, and Pain. 3) Working on an EM immungold studies of the brain of related to trafficking of glutamate and GABA receptors in mice with researchers in the laboratory of Richard Huganir of Johns Hopkins School of Medicine in Baltimore. 4) Working with researchers in the laboratory of Dr. Paul Worley of Johns Hopkins School of Medicine in Baltimore on EM immunogold studies of several proteins related to protein trafficking in neurons, including distributions in normal rats and knock-in mice. One paper involving Shank3 and NMDA receptors in a mouse model of Autism is now published in the journal Cell. Another paper is near acceptance in the journal Cell and concerns APP, presenilin, and Arc trafficking related to Alzheimers disease. 5) Worked with Dr. Jeff Erickson of LSU Health Sciences Center on some EM immunogold studies of synapse plasticity. 6) Worked with Dr. Youming Lu of LSU School of Medicine on ultrastructural changes in neurons related to signaling in synaptic transmission. A manuscript is in review. So far in 2011, the lab has published (or has in press) 11 papers, half of these within NIDCD, and has several other papers with NIDCD and other labs in review or final preparation that we hope to have in press by the end of 2011.

自 2009 年 10 月 Robert Wenthold 去世以来，该实验室继续支持一些已搬到 NINDS 的 Katherine Roche 博士实验室的研究人员（Elizabeth Webber、Kai Chang、Takaaki Hirai 以及今年从 Kachar 博士实验室转来的 Chan-Ying Cheng）以及 Gail Seabold 的项目，Gail Seabold 继续主要在这里的实验室工作，尽管她的 PI 是 来自 NHLBI 的莎朗·米尔格拉姆。实验室继续根据这些研究人员的需要进行一些神经元细胞培养的准备工作。我们还监督 JEOL 1010 透射电子显微镜和相关设备，以及 Zeiss LSM710 激光共焦显微镜和 Leica Biowave 系统的维护，其中一些研究人员继续使用这些设备。此外，NIDCD 的其他研究人员的手稿审查以及各种期刊要求的审查也会定期进行。 基本上，现在的工作是与其他实验室的 PI 合作完成的，我们将在 2012 财年开始的 10 月份成为一个独立的核心设施。我们的主要职责是促进 NIDCD 中其他 PI 的项目。这里的一些合作延续了 Wenthold 博士实验室在谷氨酸受体和神经元突触相关蛋白运输方面所做的工作。然而，作为我们帮助 NIDCD 研究的承诺的一部分，该实验室越来越多地参与耳蜗发育和病理学的研究，并且在不久的将来该实验室计划开展一些有关中枢听觉功能的工作。 今年实验室作为合作者参与的具体研究项目如下： 国家疾病预防控制中心 与原 Wenthold/NIDCD 实验室的研究人员一起： 1) 与 Philip Wang 和 Stephan Brenowitz 合作完成了一项关于神经元生长锥 NMDA 受体的研究，最近提交。进行了本研究中使用的 EM 分析，并提供了培养的神经元。现已发表在《神经科学杂志》上。 2) 继续与 Gail Seabold 和 Philip Wang 一起撰写 SALM1 论文。这是一种与突触谷氨酸受体相关的粘附蛋白。进行本研究中使用的 EM 分析并提供培养的神经元。本文经《生物化学杂志》审阅后正在修订中。 （会议上也展示了海报） 3) 与 Chan-Ying Cheng 合作完成了一篇关于突触棘 FRAP 的视频方法论文，现已发表在 JoVE 杂志上。 4) 与 Chan-Ying Cheng 合作完成一篇论文，作为已发表的 PSD-95 和 SAP102 论文的附录。本附录现已发表在《交流与整合生物学》杂志上。 5) 首席研究员受邀为《神经科学家》撰写一篇关于 MAGUK 可塑性的综述论文，并得到了 Chan-Ying Cheng、Gail Seabold 和 Martin Horak 的帮助。该文件现已出版并在线发布。 国家疾病预防控制中心B 与 NIDCD 的其他人一起： 1) 与顾新星博士实验室的任大斌合作，完成了两项独立的电镜研究，研究免疫金对儿童中耳炎和成人呼吸道感染（包括慢性阻塞性肺病）相关细菌的定位。其中一项现已发表在《疫苗》杂志上。第二篇论文正在审查中。 2) 与安迪·格里菲斯博士实验室的研究人员合作，研究涉及与耳聋相关的突变体的耳蜗变化。这篇论文现已在 J. Clin 上发表。库存。 （会议上也展示了海报） 3) 在 Drs. 实验室接受培训并与 Kate Szarama 一起工作。马特·凯利和理查德·查德威克研究了与发育和疾病相关的耳蜗超微结构变化。一篇论文正在审查中，另一篇论文即将提交。 （会议上也展示了海报） 4) 与 Matt Kelley 博士实验室的 Helen May-Simera 合作（与 Kate Szarama 合作），研究发育中耳蜗中动纤毛蛋白的免疫金定位。 5) 如上所述，在 Kachar 博士的实验室工作期间，与 Chan-Ying Cheng 一起完成了 3 个项目/论文。 6) 与Tom Friedman博士实验室的研究人员一起参与了一些关于EM免疫金分布的初步研究。特别参与了关于 EM 技术的培训并与 Inna Bellyantseva 合作。 7) 与 Paul Saving 博士的实验室成员合作，研究实验小鼠视网膜表面蛋白的电镜定位。 8) 为 Bechara Kachar 博士提供毛细胞和耳蜗超微结构研究的薄片。 9) 为 Bechara Kachar 博士实验室的 Janaina Brusco 就杏仁核突触的 EM 免疫金和 3D 研究提供建议。 10) 为 Bechara Kachar 博士实验室的 Taro Fujikawa 就耳蜗中红藻氨酸谷氨酸受体的研究提供建议。 11) 为 Stephan Brenowitz 博士的实验室规划耳蜗背核的免疫电镜研究。 12) 与 Doris Wus 博士实验室一起规划内耳的电磁研究。 与其他 NIH 机构合作： 6) 与 Drs. 一起研究 sonic hedgehog 及其受体的电磁定位、修补和平滑处理。 NIA 的帕梅拉·姚和马克·马特森。第一作者的论文现已发表在《比较神经学杂志》上。我们还有另一篇第一作者手稿正在《比较神经病学杂志》上进行审查，前一篇论文的附录和其他研究正在准备中。 （会议上也展示了海报） 1) 与 Drs. 一起参与了 2 项影响神经系统的蛋白质突变（包括精神分裂症小鼠模型）的初步研究。 NIMH 的 Jie-Min Jia 和 Song Jiao（手稿正在准备中）。 2）与NICHD的Dax Hoffman实验室合作研究涉及突触整合和可塑性的钾通道辅助亚基。论文发表在《神经元》杂志上。 NIH 以外的实验室： 1) 与 Janelia Farm Research Campus (HHMI) 的 Jinhyun Kim 博士合作，研究突触间隙蛋白质的电镜免疫定位。一份手稿正在审查中。 2) 与巴尔的摩约翰·霍普金斯大学医学院的陶元祥博士合作，为 3 篇有关疼痛对脊髓神经元可塑性影响的论文进行了 EM 免疫金贡献；这些涉及其他合作者，例如博士。约翰·霍普金斯大学的理查德·胡加尼尔（Richard Huganir）和杰弗里·罗斯坦（Jeffrey Rothstein），以及乌克兰博戈莫列茨生理学研究所的 D.奥尔加·科帕奇（D. Olga Kopach）。所有 3 篇论文现已发表在《麻醉学》、《分子疼痛》和《疼痛》杂志上。 3) 与巴尔的摩约翰·霍普金斯大学医学院 Richard Huganir 实验室的研究人员一起，对小鼠大脑中与谷氨酸和 GABA 受体运输相关的大脑进行 EM 免疫研究。 4) 与巴尔的摩约翰·霍普金斯大学医学院 Paul Worley 博士实验室的研究人员合作，对与神经元中蛋白质运输相关的几种蛋白质进行 EM 免疫金研究，包括在正常大鼠和敲入小鼠中的分布。一篇涉及自闭症小鼠模型中 Shank3 和 NMDA 受体的论文现已发表在《细胞》杂志上。另一篇论文即将被《Cell》杂志接受，涉及与阿尔茨海默病相关的 APP、早老素和 Arc 转运。 5）与LSU健康科学中心的Jeff Erickson博士合作进行一些突触可塑性的EM免疫金研究。 6) 与LSU医学院的Youming Lu博士合作研究与突触传递信号相关的神经元超微结构变化。一份手稿正在审查中。 2011 年到目前为止，该实验室已发表（或已出版）11 篇论文，其中一半是 NIDCD 内部的，还有几篇由 NIDCD 和其他实验室进行审查或最终准备的其他论文，我们希望在 2011 年底前出版。