COBRE: LSU HSC: CELL & MOLECULAR CORE

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。细胞和分子核心主任：Emel Songu Mize，博士和Stephen M. Lanier，博士这个核心有三个主要的支持任务：细胞培养；蛋白质组学;和成像。该设施的工作人员是Marella Kuriakose， M.S., Jesse Guidry， B.S.（讲师）和Olha Nichols， M.S.细胞培养核心为COBRE研究人员提供原代培养。这包括大鼠主动脉平滑肌细胞(使用者：dr。Catling, Park, Wu, Songu Mize, Park)，小鼠巨噬细胞培养（用户：Dr. Boulares），大鼠新生心脏细胞培养（用户：Dr. Boulares）。Park和Wu)， HUVEC培养(用户：dr .；Park and Boulares)。除此之外，细胞培养核心设施还开发了小鼠肠系膜平滑肌细胞的其他细胞系。Lucchesi, Catling, Songu-Mize)，胚胎成纤维细胞（Boulares博士），并正在培养成年大鼠心肌细胞(dr。Lucchesi, Wu, Songu-Mize)。奥尔加·尼科尔斯（Olga Nichols）拥有共聚焦显微镜、组织学和组织病理学方面的专业知识，为了满足COBRE教师不断增长的需求，她于去年加入了COBRE，并分担了维护和向研究人员提供培养细胞以及监督核心成像部分的职责。COBRE的调查人员每周都会收到一份关于可用细胞系、传代数、冷冻供应和繁荣培养的报告。他们依次提出下一周的要求。Kuriakose女士为每位调查员提供他们需要的东西。2004年7月，蛋白质组学核心安装了一台Ettan斑点处理工作站。该仪器可自动处理蛋白质组学样品，从2D凝胶到最终的质谱处理。2005年4月，分子核心还上线了一台IcylerIQ实时PCR机和一台Experion电泳站，用于分析DNA、RNA和蛋白质。Jesse Guidry通过合作继续开发和促进蛋白质组学项目（儿童医院的磷蛋白质组LTQ分析，堪萨斯城密苏里大学的人类血清SELDI分析）。血管平滑肌表征的工作仍在继续。此外，该核心还与COBRE研究人员合作，协助融合蛋白富集，肾动脉，脑蛋白质组学，凝集素，磷酸化蛋白质组学，相互作用组学等蛋白质组学分析。1) Hamid Boulares, Karine Oumouna-Benachour， Manika Das和Kirk Hutchinson：在动脉粥样硬化小鼠模型中，PARP-1抑制促进动脉粥样硬化斑块稳定性因子，并在体外防止血管细胞死亡。血管生物学与医学2005：从分子到人；[2]张志强，李建军，李建军，等。大鼠脑缺血缺血再灌注后血管内皮细胞TRPC表达及钙离子迁移的研究进展。[3]李建军，李建军，李建军，等。大鼠血管平滑肌细胞TRPC4蛋白和基因表达的研究进展。提交给美国心脏协会，高血压协会，2005年4)凯文·洛德，瑞安·里德，艾丽莎·希克斯，帕梅拉·卢切西和库尔特·瓦纳。氧化应激在甲基苯丙胺诱导的心功能障碍中的作用。重点：心脏疾病和再生的分子生物学，2005年4月3-8日。5)西蒙小提琴，杰西·古德里，周卓文，史蒂芬·M·拉尼尔。与M2毒蕈碱受体细胞内第三环络合的辅助蛋白的鉴定。6) RE Reed, HH Hughie LJ Dell？Italia和PA Lucchesi。过氧亚硝酸盐的产生增加是左室细胞外基质重塑的关键介质，以应对急性容量过载。重点：心脏疾病和再生的分子生物学，2005年4月3-8日。4)，马丽娟，马丽娟，马丽娟。MEK1支架蛋白MP1通过整合PAK1和Rho信号调控细胞扩散。摩尔。细胞。医学杂志。，出版。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Cell and Molecular Core Directors: Emel Songu Mize, Ph.D. and Stephen M. Lanier, Ph.D. This core has three major support missions: Cell Culture; Proteomics; and Imaging. The facility is staffed by Marella Kuriakose, M.S., Jesse Guidry, B.S. (Instructor)and Olha Nichols, M.S. CELL CULTURE The core supplies primary cultures for COBRE investigators. This includes rat aortic smooth muscle cells (users: Drs. Catling, Park, Wu, Songu Mize, Park), mouse macrophage cultures (user: Dr. Boulares), rat neonatal cardiac cell cultures (user: Drs. Park and Wu), HUVEC cultures (users: Drs. Park and Boulares). In addition to the above, the Cell Culture core facility developed additional cell lines of mouse mesenteric smooth muscle cells (Drs. Lucchesi, Catling, Songu-Mize), embryonic fibroblasts (Dr. Boulares) and is in the process of establishing adult rat cardiac myocytes in culture (Drs. Lucchesi, Wu, Songu-Mize). In order to respond to the expanding needs of the COBRE faculty Ms. Olga Nichols, who has expertise in confocal microscopy, histology and histopathology, came on board last year and is sharing the expanding responsibilities of maintaining and providing cultured cells to the investigators and overseeing the Imaging component of the Core. The COBRE investigators are provided a weekly report of available cell lines, passage numbers, both the frozen supplies and thriving cultures. They in turn make their request for the following week. Ms. Kuriakose delivers to each investigator what they need. PROTEOMICS AND IMAGING In July 2004, the proteomics core installed an Ettan Spot Handling Workstation. This instrument automates the processing of proteomic samples from 2D gels unto the ultimate processing via mass spectrometry. In April 2005, the molecular core also brought online an IcylerIQ real-time PCR machine and an Experion Electrophoresis station for the analysis of DNA, RNA and protein. Jesse Guidry continues to develop and foster proteomics projects through collaboration (LTQ analysis of phosphoproteome at Children's Hospital, SELDI analysis of human serum at University of Missouri, Kansas City). Work continues in the vascular smooth muscle characterization. In addition, the core is also aiding in the proteomic analysis of fusion protein enrichment, renal artery, brain proteomics, galectins, phosphoproteome, interactomes and others in conjunction with COBRE investigators. A list of abstracts and a manuscript that resulted from the use of Cell Culture Core cell lines is attached: Abstracts 1) Hamid Boulares, Karine Oumouna-Benachour, Manika Das, and Kirk Hutchinson: PARP-1 inhibition promotes factors of atherosclerotic plaque stability in a mouse model of atherosclerosis and prevents death of vascular cells in vitro. Vascular Biology and Medicine 2005: From Molecules to Man; Chicago June 16-19, 2005 2) Hoffmann SE, Zhang Z, Songu-Mize E: Effect of cyclic stretch on TRPC expression and calcium mobilization. Experimental Biology, April 2-6, 2005 San Diego. 3) Hoffmann SE, Kuriakose M, Songu-Mize E: Effect of cyclic stretch on TRPC4 protein and gene expression in vascular smooth muscle cells. Submitted to American Heart Association, High Blood Pressure Council, 2005 4) Kevin Lord, Ryan Reed, Alissa Hicks, Pamela Lucchesi and Kurt Varner. Role of Oxidative stress in methamphetamine-induced cardiac dysfunction. Keystone: Molecular Biology of Cardiac Diseases and Regeneration April 3-8th, 2005. 5) Violaine Simon, Jessie Guidry, Chau-Wen Chou and Stephen M Lanier. Identification of accessory proteins complexing with the third intracellular loop of M2 muscarinic receptors. Experimental Biology, April 2-6, 2005 San Diego. 6) RE Reed, HH Hughie LJ Dell?Italia and PA Lucchesi. Increased peroxynitrite production is a key mediator of LV extracellular matrix remodeling in response to acute volume overload. Keystone: Molecular Biology of Cardiac Diseases and Regeneration April 3-8th, 2005. Manuscripts: 4) Pullikuth, A. K., McKinnon, E., Schaeffer, H. J., and Catling, A. D. The MEK1 scaffolding protein MP1 regulates cell spreading by integrating PAK1 and Rho signals. Mol. Cell. Biol., in press.