Molecular Mechanisms Of Synaptic Transmission

根据这些研究人员的需要，该实验室继续进行一些神经元细胞培养准备工作。我们还监督JEOL 1010透射电子显微镜和相关设备的维护，以及蔡司LSM710激光共聚焦显微镜和徕卡BioWAVE系统的维护，其中一些研究人员和邻近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Ya-Xian Wang在2011财年开始了几个项目，这些项目在去年的年报中进行了描述，并在2012财年开始了一些新的项目。这些都在新的先进成像核心的年度报告中进行了详细描述。然而，我们也将相关的出版作品也包括在本报告中，因为它们中的大多数都是在2011财年开始的。另外，我们将在这里具体总结盖尔·希伯德、张凯、嬴政的工作。 Gail Seabold在2012财年工作期间完成了两个项目。她完成了关于突触黏附样分子SALM1的主要论文的修订，以及二亮氨酸和PDZ结合基序在其在海马神经元运输中的作用。这项工作是与罗纳德·彼得里亚、罗伯特·温托尔德和王亚贤以及实验室前成员菲利普·王和张凯合作完成的。它终于在2012年初发表在《生物化学杂志》上，并被选为本期的封面照片。她还与罗纳德·佩特里亚(Ronald Petralia)(和马丁·霍拉克)合作撰写了一篇关于谷氨酸受体贩运和可塑性的大型综述论文，并于2012年春天在网上发表。盖尔还与罗纳德·佩特里亚合作进行了一些初步的新研究，这些研究正在进行中。 此外，盖尔在实验室中发挥了很大的作用，帮助在实验室中通过抗体、DNA质粒和其他材料进行组织和筛选，以及在实验室中提供帮助。 张凯使用了分子生物学、细胞生物学和组织学技术，对几种代谢性谷氨酸受体mGluR5转基因小鼠的黑色素瘤进行了特征分析(Choi K.Y和Chang K.，等人)。PNAS108(37)：15219-15224,2011年)。在下一阶段对mGluR5诱导的黑色素瘤的研究中，他专注于mGluR5异位表达导致黑素细胞肿瘤发生的分子机制。他在这方面的主要成就包括：解开了转导mGluR5的黑素细胞ERK1/2激活的信号通路，鉴定了参与mGluR5诱导的黑色素瘤形成的蛋白质(可能用于诊断和治疗)，阐明了不同mGluR5结构域的功能并证明了胞外结构域的脱落与细胞增殖有关，建立了可用于体外肿瘤发生分析的mGluR的转化实验和稳定的mGluR细胞系，并研究了可能与mGluR的C-尾结合并作为辅助受体发挥功能的非受体酪氨酸激酶。 2011年，嬴政与罗纳德·佩特里亚和王亚贤合作发表了三篇论文，其中包括与温特霍尔德和卡查尔博士合作发表的两篇研究论文。这些是这三人在2010年发表的一篇论文的后续论文。其中一篇发表在《视觉实验杂志》上，描述了pEGFP载体在培养的海马神经元脊椎中光漂白后的荧光恢复。另一项是超分辨率显微镜研究，发现MAGUK(突触支架蛋白)、SAP102和PSD-95的定位略有不同，这一差异需要超分辨率才能识别。第三篇是关于支架蛋白MAGUK及其在突触发育和可塑性中的作用的综述文章。这篇文章发表在《神经学家》杂志上，也被用作该期的封面照片。 在Chan-Ying加入Katherine Roche博士在NINDS的实验室后，她开始研究两种AMPA受体结合蛋白Gamma 8和Cornichon 2的运输和功能。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实验室，Gamma 8 KO小鼠由Dr.Roger Nicolls Group赠送。Chan-Ying正在使用成像和生物化学方法进行Gamma 8和Cornichon 2项目。 她最近发表了一篇Cornichon 2的论文，这篇论文已于上个月由Nicoll小组提交。她是那篇论文的第三作者。 同样在今年，陈莹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，她已经审阅了10篇手稿。其中一半是由罗纳德·彼得利亚推荐的，她帮助审查了她在评论中的评论。她评论的期刊包括：2011年--《分子细胞生物学杂志》，《科学中国》，《生物学前沿》。2012-Plos One*(2篇论文)，交流与整合生物学，神经药理学，国际发育神经科学杂志*(2篇论文)，药理学报。 陈颖将继续与凯瑟琳·罗氏合作，直到2013财年。

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.