Molecular Mechanisms Of Synaptic Transmission

突触传递的分子机制

基本信息

批准号：
8565491
负责人：
David Robinson
金额：
$ 38.42万
依托单位：
NATIONAL INSTITUTE ON DEAFNESS AND OTHER COMMUNICATION DISORDERS
依托单位国家：
美国
项目类别：
财政年份：
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/8565491
关键词：
AMPA Receptors Adhesions Annual Reports Antibodies Axon Binding Binding Proteins Biochemistry Biological Assay Biology Cell Adhesion Molecules Cell Culture Techniques Cell Line Cell Proliferation Cellular biology China Cochlea Collaborations Communication Dendrites Development Diagnosis Disease Ectopic Expression Electron Microscope Equipment Excitatory Synapse Fluorescence Recovery After Photobleaching Gifts Glutamate Receptor Hair Cells Hippocampus (Brain)Histology Image In Vitro International Investigation Journals Knockout Mice Laboratories Lasers Lead Location MAPK3 gene Maintenance Manuscripts Metabotropic Glutamate Receptors Methods Microscope Microscopy Molecular Molecular Biology Techniques N-Methylaspartate National Institute of Neurological Disorders and Stroke National Institute on Deafness and Other Communication Disorders Neuraxis Neurons Neuropharmacology Neurosciences Paper Phase Preparation Process Protein Tyrosine Kinase Proteins Published Comment Publishing Regulation Reporting Research Research Personnel Resolution Role Scaffolding Protein Science Signal Pathway Supporting Cell Synapses Synaptic Transmission System Tail Transgenic Mice Vertebral column Visual Work follow-up frontier interest kainate melanocyte melanoma member nervous system disorder neuropathology plasmid DNA receptor receptor binding research study scaffold synaptic function trafficking transmission process tumorigenesis vector

项目摘要

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 continued to be used here by some of these researchers, and others from adjacent NIDCD labs, in FY12. Ronald Petralia and Ya-Xian Wang continued on several projects begun in FY11 and which were described in last years annual report, and began some new ones in FY12. These are described in detail in the annual report of the new Advanced Imaging Core. However, we have included the associated published works here also in this report, since most of them were begun in FY11 in this project. Otherwise, we will summarize here specifically the work of Gail Seabold, Kai Chang, and Chan-Ying Zheng. Gail Seabold worked to finish two projects while she was here in FY12. She completed the revisions of her major paper on the synaptic adhesion-like molecule, SALM1, and the role of dileucine and PDZ-binding motifs in its trafficking in hippocampal neurons. This work was a collaboration with Ronald Petralia, Robert Wenthold, and Ya-Xian Wang, along with former members of the laboratory, Philip Wang and Kai Chang. It finally was published in the Journal of Biological Chemistry in early 2012, and was featured for the cover photo of the issue. She also worked with Ronald Petralia (and Martin Horak) on a large review paper on glutamate receptor trafficking and plasticity that was published on-line in the Spring of 2012. Gail also worked with Ronald Petralia on some preliminary new studies that are in progress. In addition, Gail was of great benefit in the laboratory, helping to organize and cull through antibodies, DNA plasmids, and other materials in the laboratory, as well as helping out in the laboratory in general. Kai Chang has used techniques of molecular biology, cell biology and histology, to characterize melanomas from several strains of transgenic mice for the metabotropic glutamate receptor, mGluR5 (Choi K.Y and Chang K., et al. PNAS 108 (37): 15219-15224, 2011). In the next phase of study on mGluR5-induced melanoma, he focused on the molecular mechanisms that lead to tumorigenesis of melanocytes by mGluR5 ectopic expression. His major accomplishments in this include: unraveled the signal pathways of ERK1/2 activation in mGluR5-transfected melanocytes, characterized proteins involved in melanomagenesis induced by mGluR5 (potentially useful in diagnosis and therapy), elucidated the function of different mGluR5 domains and showed that ectodomain shedding was associated with cell proliferation, established a melan-a transforming assay and stable melan-a cell lines expressing mGluRs, useful for in vitro tumorigenesis assays, and studied non-receptor tyrosine kinases that may bind to the C-tail of mGluRs and function as co-receptors. Chan-Ying Zheng had three papers with Ronald Petralia and Ya-Xian Wang during 2011, including 2 research papers published in collaboration with Drs. Wenthold and Kachar. These were follow-up papers to one published by these three in 2010. One was a description of fluorescence recovery after photobleaching of pEGFP vector in spines of cultured hippocampal neurons, published in the Journal of Visual Experiments. Another was a study on super resolution microscopy that revealed the slightly different localization of the MAGUKs (synaptic scaffolding proteins), SAP102 and PSD-95 a difference that required super resolution to identify. The third was a review article on the scaffolding proteins, MAGUKs, and their role in synaptic development and plasticity. This was published in the Neuroscientist, and also was used for the cover photo for that issue. After Chan-Ying joined the laboratory of Dr. Katherine Roche in NINDS, she began studying the trafficking and function of two AMPA receptor binding proteins, Gamma 8 and Cornichon 2. These projects are ongoing. Recently, she received two knockout (KO) mice. The Gamma 2 KO mice were purchased from Jackson Laboratories and Gamma 8 KO mice were gifts from Dr. Roger Nicolls group. Chan-Ying is working on Gamma 8 and Cornichon 2 projects using imaging and biochemistry methods. She recently contributed to a Cornichon 2 paper, which has been submitted by the Nicoll group last month. She is the third author of that paper. Also this year, Chan-Ying has been actively reviewing papers submitted to journals by other research groups. She has reviewed 10 manuscripts since November 2011. Half of them were referred by Ronald Petralia, who helped to review her comments in her reviews. The journals that she reviewed include: 2011-- Journal of Molecular Cell Biology, Science China, Frontiers in Biology. 2012-Plos One*(2 papers), Communicative & Integrative Biology, Neuropharmacology, International Journal of Developmental Neuroscience*(2 papers), Acta Pharmacologica Sinica. Chan-Ying will continue her work with Katherine Roche into FY13.

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，其中一些研究人员以及来自邻近 NIDCD 实验室的其他研究人员在 2012 财年继续使用这些设备。 Ronald Petralia 和王亚贤继续开展 2011 财年开始并在去年年度报告中描述的几个项目，并在 2012 财年开始一些新项目。这些在新的高级成像核心的年度报告中都有详细描述。然而，我们也将相关已发表的作品纳入本报告中，因为其中大部分是 2011 财年在该项目中开始的。除此之外，我们将在这里具体总结 Gail Seabold、Kai Chang 和 Chan-Ying Cheng 的工作。 2012 财年，盖尔·西博尔德 (Gail Seabold) 在此期间完成了两个项目。她完成了关于突触粘附样分子 SALM1 以及二亮氨酸和 PDZ 结合基序在其海马神经元运输中的作用的主要论文的修订。这项工作是与 Ronald Petralia、Robert Wenthold 和 Ya-Xian Wang 以及实验室前成员 Philip Wang 和 Kai Chang 合作完成的。最终于2012年初发表在《生物化学杂志》上，并作为该期封面照片。她还与 Ronald Petralia（和 Martin Horak）合作撰写了一篇关于谷氨酸受体运输和可塑性的大型评论论文，该论文于 2012 年春季在线发表。Gail 还与 Ronald Petralia 合作进行了一些正在进行的初步新研究。此外，盖尔在实验室也大有裨益，帮助组织和剔除实验室中的抗体、DNA质粒和其他材料，并在实验室中提供一般帮助。 Kai Chang 使用分子生物学、细胞生物学和组织学技术，对代谢型谷氨酸受体 mGluR5 的几种转基因小鼠品系的黑色素瘤进行了表征（Choi K.Y 和 Chang K. 等人。PNAS 108 (37): 15219-15224 ，2011）。在mGluR5诱导黑色素瘤的下一阶段研究中，他重点研究mGluR5异位表达导致黑色素细胞肿瘤发生的分子机制。他在这方面的主要成就包括：揭示了 mGluR5 转染的黑色素细胞中 ERK1/2 激活的信号通路，表征了 mGluR5 诱导的黑色素瘤发生中涉及的蛋白质（可能在诊断和治疗中有用），阐明了不同 mGluR5 结构域的功能，并表明胞外域脱落与细胞增殖相关，建立了 melan-a 转化测定和表达 mGluRs 的稳定 melan-a 细胞系，有用用于体外肿瘤发生测定，并研究了可能与 mGluR 的 C 尾结合并充当辅助受体的非受体酪氨酸激酶。 2011 年，Chan-Ying Cheng 与 Ronald Petralia 和 Ya-Xian Wang 合作发表了三篇论文，其中包括 2 篇与 Drs.温特霍尔德和卡查尔。这些是这三人于 2010 年发表的一篇论文的后续论文。其中一篇描述了培养的海马神经元棘中 pEGFP 载体光漂白后的荧光恢复，发表在《视觉实验杂志》上。另一项是超分辨率显微镜研究，揭示了 MAGUK（突触支架蛋白）、SAP102 和 PSD-95 的定位略有不同，这种差异需要超分辨率才能识别。第三篇是一篇关于支架蛋白 MAGUK 及其在突触发育和可塑性中的作用的评论文章。这篇文章发表在《神经科学家》杂志上，也被用作该期的封面照片。 Chan-Ying 加入 NINDS Katherine Roche 博士的实验室后，开始研究两种 AMPA 受体结合蛋白 Gamma 8 和 Cornichon 2 的转运和功能。这些项目正在进行中。最近，她收到了两只基因敲除（KO）小鼠。 Gamma 2 KO小鼠购自Jackson Laboratories，Gamma 8 KO小鼠由Roger Nicolls博士团队赠送。 Chan-Ying 正在使用成像和生物化学方法开展 Gamma 8 和 Cornichon 2 项目。她最近为 Cornichon 2 论文做出了贡献，该论文已由 Nicoll 小组上个月提交。她是该论文的第三作者。今年，Chan-Ying 一直在积极审查其他研究小组提交给期刊的论文。自 2011 年 11 月以来，她审阅了 10 篇手稿。其中一半是罗纳德·佩特拉利亚 (Ronald Petralia) 推荐的，他帮助审阅了她评论中的评论。审阅的期刊包括：2011--Journal of Molecular Cell Biology、Science China、Frontiers in Biology。 2012-Plos One*（2篇），交流与整合生物学，神经药理学，国际发育神经科学杂志*（2篇），药理学报。 Chan-Ying 将在 2013 财年继续与 Katherine Roche 合作。

项目成果

期刊论文数量（20）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

NMDA receptors interact with flotillin-1 and -2, lipid raft-associated proteins.

NMDA 受体与 flotillin-1 和 -2（脂筏相关蛋白）相互作用。

DOI：
发表时间：
2009-04-17
期刊：
影响因子：
3.5
作者：
Swanwick, Catherine Croft;Shapiro, Marietta E;Yi, Zhaohong;Chang, Kai;Wenthold, Robert J
通讯作者：
Wenthold, Robert J

Differential localization of SAP102 and PSD-95 is revealed in hippocampal spines using super-resolution light microscopy.

使用超分辨率光学显微镜揭示了海马棘中 SAP102 和 PSD-95 的差异定位。

DOI：
发表时间：
2011-01
期刊：
影响因子：
0
作者：
Zheng, Chan;Wang, Ya;Kachar, Bechara;Petralia, Ronald S
通讯作者：
Petralia, Ronald S

Trafficking of the NMDAR2B receptor subunit distal cytoplasmic tail from endoplasmic reticulum to the synapse.

NMDAR2B 受体亚基远端胞质尾部从内质网运输到突触。

DOI：
发表时间：
2012
期刊：
影响因子：
3.7
作者：
Standley, Steve;Petralia, Ronald S;Gravell, Manneth;Hamilton, Rebecca;Wang, Ya;Schubert, Manfred;Wenthold, Robert J
通讯作者：
Wenthold, Robert J

mGRASP enables mapping mammalian synaptic connectivity with light microscopy.

mGRASP 能够利用光学显微镜绘制哺乳动物突触连接图。

DOI：
发表时间：
2011-12-04
期刊：
影响因子：
48
作者：
Kim, Jinhyun;Zhao, Ting;Petralia, Ronald S;Yu, Yang;Peng, Hanchuan;Myers, Eugene;Magee, Jeffrey C
通讯作者：
Magee, Jeffrey C

Subcellular localization of Patched and Smoothened, the receptors for Sonic hedgehog signaling, in the hippocampal neuron.

海马神经元中声波刺猬信号受体 Patched 和 Smoothened 的亚细胞定位。