Advanced Imaging Core

先进的成像核心

• 批准号: 8940142
• 负责人: Ronald Petralia
• 金额: $ 77.71万
• 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/8940142
• 关键词: Adhesions; Affect; Aging; Amygdaloid structure; Animals; Auditory; Autophagocytosis; BAX gene; Biological; Biological Psychiatry; Brain region; Caspase; Cells; Cerebellum; Cochlea; Collaborations; Core Facility; Czech Republic; Data; Development; Electron Microscope; Electron Microscopy; Erinaceidae; Evolution; Face; France; Future; Gaucher Disease; Genetic; Goals; Grant Review; Hair Cells; Hearing; Hippocampus (Brain); Human Pathology; Image; Institutes; Interruption; Intramural Research; Ion Channel; Journals; Kainic Acid Receptors; Label; Laboratories; Left; Letters; Light Microscope; Manuscripts; Medial; Mentors; Metabotropic Glutamate Receptors; Methods; Microscope; Molecular; Molecular Genetics; Motor; Mus; Mutant Strains Mice; Myosin ATPase; N-Methyl-D-Aspartate Receptors; National Heart, Lung, and Blood Institute; National Human Genome Research Institute; National Institute of Child Health and Human Development; National Institute on Deafness and Other Communication Disorders; Neurology; Neurons; Neuropharmacology; Neurosciences; Nociception; Outer Hair Cells; Paper; Parkinson Disease; Performance; Physiology; Positioning Attribute; Postdoctoral Fellow; Preparation; Principal Investigator; Proteins; Publishing; Recommendation; Reporting; Research; Research Personnel; Research Project Grants; Research Support; Research Training; Resources; Retinal; Retinal Diseases; Role; Running; Safety; Sampling; Scanning; Scanning Electron Microscopy; Schizophrenia; Scientist; Services; Societies; Somatosensory Cortex; Specialist; Staging; Staining method; Stains; Stereocilium; Structure; Students; Synapses; Synaptic Vesicles; System; Taro Vegetable; Techniques; Time; Tissues; Training; Transmission Electron Microscopy; Ultramicrotomy; United States National Institutes of Health; Universities; Utricle structure; Variant; Work; Writing; Zebrafish; comparative; dorsal cochlear nucleus; experience; frontier; immunocytochemistry; interest; meetings; mutant; neuroimmunology; new technology; operation; posters; programs; receptor recycling; research study; ribbon synapse; tissue preparation; tomography; trafficking; transmission process

Ronald S. Petralia and Ya-Xian Wang have run the Advanced Imaging Core facility of NIDCD since October 2011. This core is dedicated primarily to training and collaborating with other scientists in NIDCD in studies utilizing electron microscopy (EM). Also, to a limited extent, some work is done with other NIH institutes or outside NIH. In addition, Ronald Petralia regularly reviews papers for professional journals and reviews many various documents for NIDCD PIs, reviews grants, writes review papers and letters of recommendation, serves on several NIH committees and helps coordinate lab+office safety for NIDCD (including the Clean Sweep this year). This year, we experienced a significant interruption (for about 2 months) due to the move from building 50 to 35A. In addition, we switched to a new, advanced transmission electron microscope that includes EM Tomography. Installation and set-up of this EM took considerable time. And this has entailed extensive, ongoing training on the new microscope system and methods. Projects (RCDC=Neuro, Aud Sys): NIDCD: Matt Kelley: We worked with fellow, Helen May-Simera (now in NEI), on the ciliary proteins, BBS8 and IFT20, as part of a study on 'ciliary' proteins and their association with the PCP protein Vangl2; The Core has been performing EM immunogold labeling plus EM on the developing mouse cochlea. For this study, the Core also has been involved in the extensive cochlea tissue preparation, prepared previously in collaboration with Kate Szarama. A manuscript received favorable reviews and is currently in revision for the journal Development. We also are performing studies on another ciliary protein. Finally, we recently started planning a study with Weise Chang. (1F-Dev-Aud Sys) Bechara Kachar: The Core recently has been training the new fellow, Robstein Chidavaenzi, on electron microscope tissue preparation, ultramicrotomy, immunolabeling, and electron microscope operation. The Core also has been involved in several projects either recently published or in preparation, with Dr. Kachars lab. The first was with Taro Fujikawa on the kainate receptors of the outer hair cells and their afferent terminals; it involved previous training and research. The findings were published in Hearing Research this year. The second project was with Janaina Brusco, on EM ultrastructure of synapses in the posterodorsal medial amygdala. This paper was published in the Journal of Comparative Neurology this year. The third project is with Seham Ebrahim. In late 2013, we performed tissue preparation, ultramicrotomy, immunolabeling, staining, and EM of auditory and vestibular tissue, especially looking at the structure of normal stereocilia and comparing this to the structure of stereocilia in espin mutant mice. The Core also has been working with Dr. Kachar on studies involving the hair cell synaptic ribbon, especially of a mutant mouse for Vglut3, and we hope to develop this further with the use of EM Tomography. The Core also gives help with other projects of the lab. (1G&2B-Per Aud/Vest S&F) Tom Friedman/Inna Belyantseva: The Core currently is training Inna Belyantseva on use of the new electron microscope and related techniques. (1G&2B-Per Aud/Vest S&F) Stephan Brenowitz: The Cores previous extensive work with postdoctoral fellows from Dr. Brenowitzs laboratory, Shan He and Milos Sedlacek (including mentoring of a summer student) was terminated when Dr. Brenowitz left NIH in late 2013, to start a new position at Janelia. However, for the study with Dr. He, on ion channels of the cartwheel cell synapses from the dorsal cochlear nucleus, we did publish a paper in The Journal of Neuroscience this year. (1A-Cen Aud S&F) Lisa Cunningham/Lindsey May/Andrew Breglio: The Core has been working extensively now on 2 separate projects for this laboratory. We have been performing morphological analyses of utricle exosomes with EM for a study with Lindsey May. As a part of this study, the Core helped to mentor a summer student, Tyra Martinez; she presented a summer poster of her findings in August. Also, recently, we performed EM Tomography on EM material from this study. We also have been preparing samples of cochlea for Serial Block Face Imaging using a scanning electron microscope with a built-in Ultramicrotome. (2B-Vest S&F) Katie Kindt: The Core has been working on structural changes in ribbon synapses from Zebrafish hair cells. Recently, this has included study of these synapses using EM Tomography. Yong Zeng/Vijay Camasamudram/Hongman Song/Ron Bush NIDCD/NEI: The Core has prepared samples and examined retinal tissue with EM for 2 different studies on retinal diseases, involving Yong/Vijay and Hongman, respectively. The Core currently has worked with Gail Seabold (formerly NIDCD/currently in the Graduate Partnerships Program/OITE), on synaptic adhesion-like molecule 1 (SALM1) trafficking in neurons in collaboration with a lab in France and involves a SALM1-associated human pathology (manuscript in preparation). Other NIH institutes: NIA /Mark Mattson/Pamela Yao lab: Projects include: endocytic accessory proteins in neurons; another looks at sonic hedgehog in synapse development and function. We will present a poster at the Society for Neuroscience National Meeting on this in November. In addition, we published 2 review papers (Petralia is first author on both), on aging synapses and the evolution of aging mechanisms in animals. The Core continues to be involved in research studies with this group. NICHD Chris McBain/Ken Pelkey/Megan Wyeth lab: The Core worked on a project looking at the role of a kainate receptor auxiliary protein in hippocampal circuitry. This involved preparation of tissue (KO and WT mice), immunogold labeling, and EM, as well as training of the fellow, Megan. Earlier this year, we published a paper on this in The Journal of Neuroscience. Currently, we are working on another research project related to this topic. NHGRI Ellen Sidransky/Nahid Tayebi: Currently, we are working on an ultrastructural study of changes in different regions of the brain in mice with combinations of Parkinsons and Gauchers diseases. NHLBI John Hammer lab: Last year, we studied the localization of the motor protein, myosin My18A in the cerebellum, utilizing EM methods. It is not clear when this study will be completed. NIHMS: Last year we worked on a study with Song Jiao and Zheng Li, on autophagy and synaptic vesicles affected by BAD-BAX-caspase; we expect that this study will be completed and published in the near future. Outside NIH: We published a paper with Paul Worley, Jay Baraban and Irving Reti of Johns Hopkins University, on Narp in nociception in press in the Journal of Neuroimmunology. Others include: with Gavin Rumbaugh (Scripps Institute) on SynGAP mutant effects on synapses of somatosensory cortex (associated with schizophrenia), in press in Biological Psychiatry; one with Jaroslav Blahos of the Institute of Molecular Genetics in the Czech Republic examining associations between 2 variants of the metabotropic glutamate receptor, mGluR1, in press in Neuropharmacology; and one with Houhui Xia from LSU on the neuronal phosphoproteome. Finally, we also have a paper with Nathalie Sans of the University of Bordeaux in France in the final stage of acceptance for Cell Reports, concerning the role of Scribble1 in NMDA receptor recycling. In addition, we recently submitted an invited review paper on NMDA receptor trafficking with Nathalie Sans and Martin Horak (Czech Republic), to Frontiers in Neuroscience. Finally, we are working on a new project with Martin on NMDA receptor trafficking.

自2011年10月以来，Ronald S. Petralia和Ya-xian Wang已运行了NIDCD的先进成像核心设施。该核心主要致力于使用电子显微镜（EM）的NIDCD的其他科学家进行培训和与其他科学家合作。同样，在有限的范围内，与其他NIH机构或NIH以外进行了一些工作。此外，罗纳德·佩特里亚（Ronald Petralia）定期审查专业期刊的论文，并审查了许多有关NIDCD PIS的各种文件，审查赠款，撰写审查论文和推荐信，在几个NIH委员会上任职，并帮助协调NIDCD的LAB+办公室安全（包括今年的清洁扫描）。 今年，由于从50号建筑物转移到35A，我们经历了重大中断（约2个月）。此外，我们改用了包括EM断层扫描的新的高级传输电子显微镜。此EM的安装和设置花费了大量时间。这需要对新显微镜系统和方法进行广泛的，正在进行的培训。 项目（rcdc = neuro，aud sys）： NIDCD： 马特·凯利（Matt Kelley）：我们与Helen May-Simera（现为NEI），纤毛蛋白，BBS8和IFT20一起工作，作为“纤毛”蛋白的研究的一部分及其与PCP蛋白vangl2的关联。核心一直在发育中的小鼠耳蜗上执行EM免疫金标签加上EM。在这项研究中，核心还参与了以前与Kate Szarama合作准备的广泛耳蜗组织制备。手稿收到了有利的评论，目前正在修订《期刊开发》。我们还在对另一种睫状蛋白进行研究。最后，我们最近开始与Weise Chang计划一项研究。 （1f-dev-aud sys） Bechara Kachar：核心最近一直在训练新的研究员Robstein Chidavaenzi，该研究人员介绍了电子显微镜组织制备，超大切开术，免疫标记和电子显微镜操作。核心还参与了最近发布或准备的几个项目，并与Kachars博士实验室一起参与。首先是在外毛细胞及其传入末端的海藻酸盐受体上使用芋头藤川。它涉及以前的培训和研究。研究结果于今年发表在听力研究中。第二个项目是Janaina Brusco，关于后内侧杏仁核突触的EM超微结构。本文发表在今年的《比较神经病学杂志》上。第三个项目是与塞汉姆·埃布拉希姆（Seham Ebrahim）一起。在2013年底，我们进行了组织制备，超细切开术，免疫标记，染色以及听觉和前庭组织的EM，尤其是查看正常立体胶质的结构，并将其与Espin突变小鼠中立体尾依性结构进行了比较。核心还一直与Kachar博士合作研究涉及毛细胞突触色带的研究，尤其是用于VGLUT3的突变小鼠，我们希望通过使用EM断层扫描进一步发展这一点。核心还为实验室的其他项目提供了帮助。 （1G和2b/aud/vest s＆f） 汤姆·弗里德曼（Tom Friedman）/inna belyantseva：目前，核心是使用新电子显微镜和相关技术训练inna belyantseva。 （1G和2b/aud/vest s＆f） 斯蒂芬·布罗诺维茨（Stephan Brenowitz）：核心先前与布雷诺维茨（Brenowitzs）实验室博士的博士后研究员（Shan He）和米洛斯·塞德拉斯克（Milos Sedlacek）（包括夏季学生的指导）在2013年底离开NIH，以开始在珍妮莉亚（Janelia）的新职位时，终止了核心。但是，对于与He博士的研究，在背侧耳蜗核心的Cartwheel细胞突触的离子通道上，我们确实在今年的神经科学杂志上发表了一篇论文。 （1A-CEN AUD S＆F） 丽莎·坎宁安（Lisa Cunningham）/林赛·梅（Lindsey May）/安德鲁·布雷格里奥（Andrew Breglio）：核心现在已经在这个实验室为两个单独的项目进行了广泛的工作。我们一直在对Lindsey May进行一项研究，对EM进行尿素外泌体的形态分析。作为这项研究的一部分，核心帮助指导了夏季学生泰拉·马丁内斯（Tyra Martinez）；她在八月份介绍了她发现的夏季海报。另外，最近，我们对这项研究的EM材料进行了EM层析成像。我们还一直在使用带有内置的超级机体的扫描电子显微镜来制备耳蜗样品，用于串行块面成像。 （2b-vest S＆F） 凯蒂·金特（Katie Kindt）：核心一直在研究来自斑马鱼毛细胞的色带突触的结构变化。最近，这包括使用EM断层扫描对这些突触的研究。 Yong Zeng/Vijay Camasamudram/Hongman Song/Ron Bush Nidcd/Nei：核心已经准备了样品并检查了EM的视网膜组织，分别涉及Yong/Vijay和Hongman的2种不同的视网膜疾病研究。 目前，该核心与Gail Seabold（以前是NIDCD/目前在研究生合作伙伴计划/OETE中）合作，与法国实验室合作，在神经元中类似突触粘附的分子1（SALM1）运输，并涉及SALM1相关的人类病理学（制备中的手稿）。 其他NIH机构： NIA /Mark Mattson /Pamela Yao实验室：项目包括：神经元中的内吞辅助蛋白；另一个看着突触发展和功能中的声音刺猬。我们将在11月在神经科学全国会议上介绍一张海报。此外，我们发表了两篇评论论文（Petralia都是第一作者），有关动物衰老机制的衰老突触的演变。核心继续参与该小组的研究。 NICHD CHRIS MCBAIN/KEN PELKEY/MEGAN WYETH LAB：核心在一个项目中工作，着眼于海马电路中的Kainate受体辅助蛋白的作用。这涉及到组织（KO和WT小鼠），免疫金标签以及EM的准备，以及对同伴梅根的培训。今年早些时候，我们在《神经科学杂志》上发表了有关此论文的论文。目前，我们正在研究与该主题有关的另一个研究项目。 NHGRI Ellen Sidransky/Nahid Tayebi：目前，我们正在研究小鼠与帕金森氏症和高乔疾病的组合的大脑不同区域变化的超微结构研究。 NHLBI John Hammer实验室：去年，我们利用EM方法研究了运动蛋白Myosin My18a的定位。目前尚不清楚该研究何时完成。 NIHMS：去年，我们与Song Jiao和Zheng Li进行了一项研究，研究了受BAX-Caspase影响的自噬和突触囊泡；我们预计这项研究将在不久的将来完成并发表。 NIH外面： 我们与约翰·霍普金斯大学的保罗·沃利（Paul Worley），杰伊·巴拉班（Jay Baraban）和欧文·埃里（Irving Reti）发表了一篇论文，载于《神经免疫学杂志》中的新闻吸引力。其他包括：与Gavin Rumbaugh（Scripps Institute）有关Syngap突变体对体感皮质突触的影响（与精神分裂症相关），在生物精神病学的印刷中；一个与捷克共和国分子遗传学研究所的Jaroslav Blahos一起研究了神经药理学中的2种变体之间的关联。一个来自LSU的Houhui Xia在神经元磷酸蛋白酶上。最后，在接受细胞报告的最后阶段，我们还与法国波尔多大学的Nathalie Sans一起发表了一篇论文，涉及Scribble1在NMDA受体回收中的作用。此外，我们最近向Nathalie Sans和Martin Horak（捷克共和国）提交了一份有关NMDA受体贩运的邀请审查论文，并向神经科学的前沿。最后，我们正在与Martin一起从事NMDA受体贩运的新项目。