Centro Nacional de Investigaciones Cardiovasculares 6 articles published in JoVE Neuroscience In vivo Positron Emission Tomography to Reveal Activity Patterns Induced by Deep Brain Stimulation in Rats Marta Casquero-Veiga1, Nicolás Lamanna-Rama2,3, Diego Romero-Miguel2,3, Manuel Desco1,2,3,4, María Luisa Soto-Montenegro2,4,5 1Centro Nacional de Investigaciones Cardiovasculares (CNIC), 2Laboratorio de Imagen Médica, Instituto de Investigación Sanitaria Gregorio Marañón, 3Departamento de Bioingeniería e Ingeniería Aeroespacial, Universidad Carlos III de Madrid, 4CIBER de Salud Mental (CIBERSAM), 5High Performance Research Group in Physiopathology and Pharmacology of the Digestive System (NeuGut), Universidad Rey Juan Carlos We describe a preclinical experimental method to evaluate metabolic neuromodulation induced by acute deep brain stimulation with in vivo FDG-PET. This manuscript includes all experimental steps, from stereotaxic surgery to the application of the stimulation treatment and the acquisition, processing, and analysis of PET images. Biology Oligomerization Dynamics of Cell Surface Receptors in Living Cells by Total Internal Reflection Fluorescence Microscopy Combined with Number and Brightness Analysis Moreno Zamai1, Antonio Trullo2, Elvira Arza1, Ugo Cavallaro3, Valeria R. Caiolfa1,2 1Unit of Microscopy and Dynamic Imaging, Centro Nacional de Investigaciones Cardiovasculares (CNIC), Madrid, Spain, 2Centro di Imaging Sperimentale, Ospedale San Raffaele, Milan, Italy, 3Unit of Gynecological Oncology Research, European Institute of Oncology IRCCS, Milan, Italy We describe an imaging approach for the determination of the average oligomeric state of mEGFP-tagged-receptor oligomers induced by ligand binding in the plasma membrane of living cells. The protocol is based on Total Internal Reflection Fluorescence (TIRF) microscopy combined with Number and Brightness (N&B) analysis. Biology Synthesis of 68Ga Core-doped Iron Oxide Nanoparticles for Dual Positron Emission Tomography /(T1)Magnetic Resonance Imaging Irene Fernández-Barahona1, Jesús Ruiz-Cabello2, Fernando Herranz1, Juan Pellico3 1Nanobiotechnology, Molecular Imaging and Metabolomics Lab, Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC), 2CIC biomaGUNE and CIBER de Enfermedades Respiratorias (CIBERES). Ikerbasque, Basque Foundation for Science, Universidad Complutense de Madrid (UCM), 3Advanced Imaging Unit, Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC), CIBER de Enfermedades Respiratorias (CIBERES) Here, we present a protocol to obtain 68Ga core-doped iron oxide nanoparticles via fast microwave-driven synthesis. The methodology renders PET/(T1)MRI nanoparticles with radiolabeling efficiencies higher than 90% and radiochemical purity of 99% in a 20-min synthesis. Immunology and Infection An In Vivo Mouse Model to Measure Naïve CD4 T Cell Activation, Proliferation and Th1 Differentiation Induced by Bone Marrow-derived Dendritic Cells Raquel Toribio-Fernandez1, Virginia Zorita1, Beatriz Herrero-Fernandez2, Jose M. Gonzalez-Granado1,2,3 1LamImSys Lab, Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC), 2LamImSys Lab, Instituto de Investigación Sanitaria Hospital 12 de Octubre (imas12), 3CIBER de Enfermedades Cardiovasculares Here, we present a protocol for the in vivo determination of naïve CD4 T cell (T cell) activation, proliferation, and Th1 differentiation induced by GM-CSF bone marrow (BM)-derived dendritic cells (DCs). In addition, this protocol describes BM and T-cell isolation, DC generation, and DC and T-cell adoptive transfer. Developmental Biology Tracing Gene Expression Through Detection of β-galactosidase Activity in Whole Mouse Embryos María José Blanco*1, Ana I.R. Learte*1, Miguel A. Marchena1, Emma Muñoz-Sáez1, María Antonia Cid1, Iván Rodríguez-Martín1, Cristina Sánchez-Camacho2,3 1Faculty of Biomedical and Health Sciences, Universidad Europea de Madrid, 2Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC), 3School of Doctoral Studies and Research, Universidad Europea de Madrid Here we describe the standard protocol for the detection of β-galactosidase activity in early whole mouse embryos and the method for paraffin sectioning and counterstaining. This is an easy and quick procedure to monitor gene expression during development that can also be applied to tissue sections, organs or cultured cells. Bioengineering Microwave-driven Synthesis of Iron Oxide Nanoparticles for Fast Detection of Atherosclerosis Juan Pellico1,2, Jesús Ruiz-Cabello1,2, Fernando Herranz1 1Advanced Imaging Unit, Centro Nacional de Investigaciones Cardiovasculares Carlos III and CIBER de Enfermedades Respiratorias, 2Universidad Complutense de Madrid Microwave technology enables extremely fast synthesis of iron oxide nanoparticles for atherosclerosis plaque characterization. The use of an aminobisphosphonate in the external side of the nanoparticle provides a fast accumulation in the atherosclerotic area.
