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.

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