Advanced Imaging Core

先进的成像核心

• 批准号: 8565586
• 负责人: Ronald Petralia
• 金额: $ 86万
• 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/8565586
• 关键词: Actins; Adhesions; Amygdaloid structure; Annual Reports; Antibodies; Auditory; Biochemistry; Biological; Brain; Calendar; Cells; Cerebellum; Cochlea; Collaborations; Core Facility; Data; Development; Doctor of Philosophy; Electron Microscope; Electron Microscopy; Equipment; Erinaceidae; Future; Genetic; Goals; Grant Review; Hippocampus (Brain); Image; Immediate-Early Genes; Immunoperoxidase Technics; Institutes; Intramural Research; Journals; Kainic Acid Receptors; Kinocilium; Label; Laboratories; Learning; Letters; Light Microscope; Mechanics; Medial; Methodology; Methods; Microscope; Microtubules; Molecular; Monomeric GTP-Binding Proteins; Motor; Mus; Myosin ATPase; National Heart, Lung, and Blood Institute; National Institute of Child Health and Human Development; National Institute of Neurological Disorders and Stroke; National Institute on Deafness and Other Communication Disorders; Neuregulins; Neurons; Neurosciences; Outer Hair Cells; Paper; Pattern; Performance; Phosphotransferases; Physiology; Pillar Cell; Postdoctoral Fellow; Preparation; Principal Investigator; Property; Proteins; Publishing; Rattus; Recommendation; Reporter; Reporting; Research; Research Personnel; Research Project Grants; Research Support; Resources; Retinal Diseases; Role; Safety; Scientist; Sensory; Services; Signal Transduction; Societies; Specialist; Structure; Study Section; Supporting Cell; Surface; Sweden; Synapses; Synaptic Transmission; Taro Vegetable; Techniques; Thyroid Hormones; Time; Tissues; Training; Transmission Electron Microscopy; United States National Institutes of Health; Ventricular; Work; Writing; dorsal cochlear nucleus; follow-up; graduate student; human disease; immunocytochemistry; interest; light microscopy; mTOR protein; meetings; microwave electromagnetic radiation; new technology; posters; postnatal; receptor; research study; symposium; tissue preparation; tomography; trafficking; transmission process

In October, 2011, Ronald S. Petralia and Ya-Xian Wang moved from the original project of Dr. Robert Wenthold, The Molecular Mechanisms of Synaptic Transmission, to the newly-established Advanced Imaging Core facility of NIDCD. This core is dedicated primarily to collaborating with and training others scientists in NIDCD in studies utilizing electron microscopy either alone or in conjunction with light microscopy. Also, to a limited extent, some work is done with other NIH institutes or outside NIH. Many of the projects reported here were continuations of those described in the FY11 Annual Report of the Molecular Mechanisms of Synaptic Transmission project, and this has been referred to also in the final (FY12) report of that project. The specific research projects (collaboration and/or training) that the Core has been involved in this year are described below. In addition to these projects, the Core generally oversees common equipment (microscopes, freezers, biological microwave oven, lockers, culture room, etc.) and space of the NIDCD PIs in building 50. Also, 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 laboratory and office safety efforts for NIDCD. Projects: In calendar year 2012 so far, the Core has 9 research papers published or in press, as well as 2 reviews. NIDCD: Matt Kelley/Richard Chadwick NIDCD labs: The Core works with fellow, Kate Szarama (Ph.D. Graduate Student), on mechanical properties of developing pillar cells in comparison to outer hair cells, in the mouse cochlea; this project has involved the Core in active research and extensive training. The first paper, on the cytoskeletal changes in actin and microtubules that underlie the developing surface mechanical properties of sensory and supporting cells in the mouse cochlea, is now published in the journal, Development. An additional related paper related is in review. A third paper is in review, and looks at hypothyroid pillar cells and the effect of thyroid hormone secretion on Fgf receptor. We now are looking at the relatively radical changes in the ultrastructure of pillar cells during early postnatal development and the functional implications of these changes. The Core performed extensive immunogold labeling and EM, while Kate was in Sweden, and this work followed up on Kates initial work with the Core on this. Now Kate has started working with the Core regularly on this project. Matt Kelley NIDCD lab: Work with fellow, Helen May-Simera, on the ciliary proteins, BBS8 and IFT20, as part of a study on 'ciliary' proteins and their association with the PCP protein Vangl2; This study began in early 2011. For this project, the Core has been performing EM immunogold labeling plus electron microscopy on both the developing mouse cochlea as well as on the ventricular zone of the rat brain, concentrating on labeling patterns in kinocilia. For this study, the Core also has been involved in the extensive cochlea tissue preparation, in collaboration with Kate Szarama. It is not clear yet how much work will be done on this project in the near future. Bechara Kachar lab: The Core has been involved in 3 projects in progress with Dr. Kachars lab during the designated time period of this report. The first is with Taro Fujikawa on the kainate receptors of the outer hair cells and their afferent terminals; it has involved both training and extensive performance of research. The second project is with Janaina Brusco, on EM ultrastructure of synapses, especially related to GABAergic terminal distribution, in the posterodorsal medial amygdala. The core recently performed additional EM studies for this. The third project is directly with Bechara and involves the examination of specially prepared auditory and vestibular tissue to be used for tomography or other special EM techniques. The Core expects to become more deeply involved in this study and also expand the versatility of the Core by learning new methodologies from Bechara. The Core also gives occasional help with the projects of other staff of Becharas lab. Tom Friedman/Inna Belyantseva lab: The Core has been training Inna and assisting her with tissue preparation and interpretation of data involving electron microscopy. Stephan Brenowitz lab: Last year, the Core performed some studies of immunogold labeling in the dorsal cochlear nucleus in relation to Stephans projects. Recently, this involvement has increased, with the Core now involved in 2 major projects with 2 postdoctoral fellows in the laboratory, Milos Sedlacek and Shan He. Shan recently presented some of this work at a Gordon Research Conference. Vijay Camasamudram/Yong Zeng/Ron Bush NIDCD/NEI lab: The Core has been preparing tissue and studying sections with EM for a study on changes in immunolocalization of a protein, in relation to treatment of a retinal disease. The Core also published a paper in the Journal of Biological Chemistry in early 2012 on a study done with Gail Seabold (Robert Wenthold (NIDCD)/Sharon Milgram (NHLBI) labs), on synaptic adhesion-like molecule 1 (SALM1) trafficking in neurons, We also have worked together on some associated studies. Other NIH institutes: NIA Mattson Lab/Mark Mattson and Pamela Yao lab: The Core has been involved in 2 projects with these scientists: one on the role of sonic hedgehog in synapse development and function in postmitotic neurons (5 papers recently published, in press, or in review, + posters presented at meetings); another looks at the role of endocytic accessory proteins, especially AP180 and Calm, in neurons. The Core recently adapted TSA (tyramide signal augmentation) to enhance EM labeling of terminals in hippocampal neuron cultures transfected with AP180 or Calm constructs. NICHD Chris McBain/Ken Pelkey/Megan Wyeth lab: Recently, the Core has begun work on a project looking at the role of Neto1/2, a kainate receptor auxiliary protein, in hippocampal circuitry. This involves work of the Core on preparation of tissue (KO and WT mice), immunogold labeling, and EM, as well as training of the fellow, Megan. We will have a poster on this at the Society for Neuroscience meeting. NICHD Andres Buonanno lab: We have collaborated in past years and now we are beginning a collaboration involving EM immunolocalization of antibodies to some neuregulin-associated proteins. NINDS Katherine Roche lab: The Core assisted a graduate student, Stephanie Payne, with interpretation of localization patterns in brain structures of a beta-gal KI reporter mouse, helping with tissue preparation, training, and interpretation of localization patterns. NHLBI John Hammer lab: We have been studying the localization of the motor protein, myosin My18A in the cerebellum, utilizing postembedding immunogold and preembedding immunoperoxidase methods. That lab will present a poster on this collaboration at the ASCB meeting. Outside NIH: 1) Johns Hopkins - Paul Worley lab: Last year the Core worked on several projects for this lab, and published 2 papers last year and a third this year (in Cell, Nat Neurosci). Now the Core is collaborating with Paul and Po Chen on a molecule called Rheb1, an immediate early gene that encodes for a small GTPase that activates a kinase that is a mammalian target of rapamycin (mTOR). 2) Gail Seabold: In addition to the published paper (J Biol Chem), Gail has worked with the Core on some subsequent studies on SALMs. We also hope to collaborate with some outside scientists looking at the role of SALMs in human disease. 3) Also, there are some earlier collaborations that were published recently (in Nat Meth, Neuron) or are in preparation or review.

2011年10月，Ronald S. Petralia和Ya-xian Wang从突触传播的分子机制Robert Gathhold博士的原始项目转移到了NIDCD的新成立的高级成像核心设施。该核心主要致力于与单独或与光显微镜结合使用电子显微镜的研究和培训NIDCD的其他科学家的研究。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。这里报告的许多项目都是突触传输项目分子机制的2011财年报告中描述的项目的延续，这在该项目的最终（FY12）报告中也被提及。下面介绍了核心参与的特定研究项目（协作和/或培训）。 In addition to these projects, the Core generally oversees common equipment (microscopes, freezers, biological microwave oven, lockers, culture room, etc.) and space of the NIDCD PIs in building 50. Also, 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 laboratory NIDCD的办公室安全工作。 项目： 到目前为止，在2012年日历年中，核心有9条研究论文发表或媒体，以及2项评论。 NIDCD： 马特·凯利（Matt Kelley）/理查德·查德威克（Richard Chadwick）NIDCD实验室：核心与凯特·萨拉玛（Kate Szarama）（博士学位研究生）一起工作，与小鼠耳蜗中的外毛细胞相比，开发柱细胞的机械性能；该项目涉及积极研究和广泛培训的核心。第一篇论文，关于肌动蛋白和微管的细胞骨架变化，这些论文构成了在小鼠耳蜗中发展的表面机械性能和支持细胞的发展表面机械性能，现已发表在《发育》杂志上。 在审查中正在审查其他相关论文。第三篇论文正在综述中，研究甲状腺柱细胞以及甲状腺激素分泌对FGF受体的影响。现在，我们正在研究产后早期发育期间柱细胞超微结构的相对根本变化以及这些变化的功能意义。当凯特（Kate）在瑞典时，核心执行了广泛的免疫金标签和EM，这项工作跟进了凯特（Kates）的初步工作，并以此为核心。现在，凯特（Kate）已开始定期与该项目的核心合作。 Matt Kelley NIDCD实验室：与Helen May-Simera，在纤毛蛋白，BBS8和IFT20上合作，作为“纤毛”蛋白的研究的一部分，及其与PCP蛋白vangl2的关联；这项研究始于2011年初。对于该项目，核心一直在发育中的小鼠耳蜗以及大鼠脑的心室中进行EM免疫质量标签以及电子显微镜，并集中在Kinocilia的标记模式上。在这项研究中，与凯特·萨拉马（Kate Szarama）合作，核心也参与了广泛的耳蜗组织制备。目前尚不清楚在不久的将来将在该项目上完成多少工作。 Bechara Kachar Lab：在本报告的指定时间段内，核心参与了Kachars Lab博士的3个项目。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。它涉及培训和广泛的研究表现。第二个项目是在后内侧杏仁核中与突触的EM超微结构（尤其是与Gabaergic末端分布有关的）。核心最近为此进行了其他EM研究。第三个项目直接与Bechara一起使用，涉及对用于断层扫描或其他特殊EM技术的特殊听觉和前庭组织的检查。核心希望通过从Bechara学习新方法来扩大核心的多功能性，并扩大了核心的多功能性。核心还偶尔会在Becharas Lab的其他工作人员的项目中提供帮助。 汤姆·弗里德曼（Tom Friedman）/inna belyantseva实验室：核心一直在训练INNA，并协助她对涉及电子显微镜的数据制备和解释。 斯蒂芬·布罗诺维茨（Stephan Brenowitz）实验室：去年，核心对背侧人工耳蜗核对斯蒂芬斯项目的影响进行了一些研究。最近，这一参与增加了，核心现在参与了2个主要项目，其中有2个博士后研究员，Milos Sedlacek和Shan He。 Shan最近在戈登研究会议上介绍了其中的一些工作。 Vijay Camasamudram/Yong Zeng/Ron Bush NIDCD/Nei Lab：核心一直在准备组织并研究EM的切片，以研究与视网膜疾病的治疗有关的蛋白质免疫定位的变化。 该核心还在2012年初在《生物化学杂志》上发表了一篇论文，该论文涉及盖尔·西伯尔德（Robert gethold（NIDCD）/莎朗·米尔格拉姆（NHLBI）实验室），对神经元的突触类分子1（salm1）在神经元中进行了类似突触的粘附分子（salm1）的研究，我们还研究了一些相关的研究。 其他NIH机构： Nia Mattson Lab/Mark Mattson和Pamela Yao Lab：核心与这些科学家一起参与了2个项目：一个关于Sonic Hedgehog在突触后突触发展和功能中的作用（5篇论文，最近发表，发表在Press或in Review中， +海报，在会议上介绍了 +海报）；另一个研究了内吞蛋白质蛋白，尤其是AP180和平静在神经元中的作用。核心最近对TSA（Tyramide信号增强）进行了调整，以增强用AP180或平静构建体转染的海马神经元培养物中末端的EM标记。 NICHD CHRIS MCBAIN/KEN PELKEY/MEGAN WYETH LAB：最近，核心已经开始在海马电路中的Kainate受体辅助蛋白Neto1/2（Kainate受体辅助蛋白）中进行研究。这涉及核心在制备组织（KO和WT小鼠），免疫标签以及EM以及梅根（Megan）的培训方面的工作。我们将在神经科学会议会议上有一张海报。 NICHD ANDRES BUONANNO LAB：我们在过去的几年中进行了合作，现在我们开始进行一项合作，涉及对某些神经调节蛋白相关蛋白的EM免疫定位。 Ninds Katherine Roche Lab：核心协助研究生Stephanie Payne，对Beta-Gal Ki Reporter Mouse的大脑结构中的本地化模式进行解释，以帮助进行组织准备，训练和解释定位模式。 NHLBI John Hammer实验室：我们一直在研究运动蛋白，肌球蛋白MY18A在小脑中的定位，利用后置IMMUNOGOLD和预填充免疫过氧化物酶方法。该实验室将在ASCB会议上为此合作提供海报。 NIH外面： 1）约翰·霍普金斯（Johns Hopkins） - 保罗·沃利（Paul Worley）实验室：去年，核心在该实验室的几个项目中工作，并于去年发表了2篇论文，今年发表了第三篇论文（在Cell，Nat Neurosci）。现在，核心正在与Paul and Po Chen合作，以一种称为Rheb1的分子，这是一个直接的早期基因，该基因编码一个小的GTPase，该基因激活了一种激酶，该激酶是雷帕霉素（MTOR）的哺乳动物靶标。 2）Gail Seabold：除了发表的论文（J Biol Chem）外，Gail还与核心合作，对萨尔姆斯的一些后续研究。我们还希望与一些外部科学家合作，研究萨尔姆在人类疾病中的作用。 3）此外，最近发布了一些较早的合作（在Nat Meth，Neuron）或正在准备或审查中。