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Pontificia Universidad Católica de Chile

9 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Use of Real-Time Functional Magnetic Resonance Imaging-Based Neurofeedback to Downregulate Insular Cortex in Nicotine-Addicted Smokers
Mohit Rana 1,2, Sergio Ruiz 1,2, Andrea Sánchez Corzo 1,2, Axel Muehleck 4, Sandra Eck 4, César Salinas 5, Francisco Zamorano 5,6, Claudio Silva 5, Massimiliano Rea 4, Anil Batra 4, Niels Birbaumer 1,7,8, Ranganatha Sitaram 1,2,9
1Departamento de Psiquiatría, Escuela de Medicina, Centro Interdisciplinario de Neurociencias, Pontificia Universidad Católica de Chile, 2Laboratory for Brain-Machine Interfaces and Neuromodulation, Pontificia Universidad Católica de Chile, 3Institute of Medical Psychology and Behavioral Neurobiology, University of Tübingen, 4Department of Psychiatry and Psychotherapy, University of Tübingen, 5Unidad de Imágenes Cuantitativas Avanzadas, Departamento de Imaágenes, Facultad de Medicina Clínica Alemana, Universidad del Desarrollo, 6División de Neurociencia, Centro de Investigación en Complejidad Social (neuroCICS), Facultad de Gobierno, Universidad del Desarrollo, 7Institute di Ricovero e Cura a Carattere Scientifico, 8Wyss Center for Bio and Neuroengineering, 9Institute for Biological and Medical Engineering, Pontificia Universidad Católica de Chile

In real-time functional magnetic resonance imaging (rtfMRI), brain activity is experimentally manipulated as an independent variable, and behavior is measured as a dependent variable. The protocol presented here focuses on the practical use of rtfMRI as a therapeutic tool for psychiatric disorders such as nicotine addiction.

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Immunology and Infection

Studying Organelle Dynamics in B Cells During Immune Synapse Formation
Jorge Ibañez-Vega *1, Danitza Fuentes *1, Jonathan Lagos 1,2, Jorge Cancino 3, María Isabel Yuseff 1
1Laboratory of Immune Cell Biology, Department of Cellular and Molecular Biology, Pontificia Universidad Católica de Chile, 2Faculty of Medicine, Pontificia Universidad Católica de Chile, 3Centro de Investigaciones en Biología Celular y Biomedicina, Facultad de Ciencia, Universidad San Sebastián

Herein we describe two approaches to characterize cell polarization events in B lymphocytes during the formation of an IS. The first, involves quantification of organelle recruitment and cytoskeleton rearrangements at the synaptic membrane. The second is a biochemical approach, to characterize changes in composition of the centrosome, which undergoes polarization to the immune synapse.

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Neuroscience

An Improved Protocol to Purify and Directly Mono-Biotinylate Recombinant BDNF in a Tube for Cellular Trafficking Studies in Neurons
Nicolás Stuardo 1,2, Guillermo Moya-Alvarado 1,3, Carolina Ramírez 1,3, Giampietro Schiavo 4, Francisca C. Bronfman 1,3
1Department of Physiology, Faculty of Biological Sciences, Pontificia Universidad Católica de Chile, 2Department of Molecular and Cellular Biology, Faculty of Biological Sciences, Center for Aging and Regeneration (CARE UC), Pontificia Universidad Católica de Chile, 3Institute of Biomedical Sciences. Faculty of Medicine and Faculty of Life Sciences, Universidad Andrés Bello, 4Department of Neuromuscular Diseases, UCL Queen Square Institute of Neurology and UK Dementia Research Institute at UCL, University College London Campus

Recombinant BDNF containing an Avi sequence (BDNFAvi) is produced in HEK293 cells in a cost-effective manner and is purified by affinity chromatography. BDNFavi is then directly mono-biotinylated with the enzyme BirA in a tube. BDNFavi and mono-biotinylated BDNFavi retain their biological activity when compared to commercially available BDNF.

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Bioengineering

Quantitative MRI of Endothelial Permeability and (Dys)function in Atherosclerosis
Begoña Lavin 1,2,3, Marcelo E. Andia 1,4,5, Prakash Saha 6, René M. Botnar 1,2,7,8, Alkystis Phinikaridou 1,2
1School of Biomedical Engineering and Imaging Sciences, King’s College London, 2BHF Centre of Excellence, Cardiovascular Division, King’s College London, 3Biochemistry and Molecular Biology Department, School of Chemistry, Complutense University, 4Radiology Department, School of Medicine, Pontificia Universidad Católica de Chile, 5ANID - Millennium Science Initiative Program - Millennium Nucleus for Cardiovascular Magnetic Resonance, 6Academic Department of Vascular Surgery, Cardiovascular Division, King’s College London, 7Wellcome Trust and EPSRC Medical Engineering Center, King’s College London, 8Escuela de Ingeniería, Universidad Católica de Chile

We have developed an accurate, non-invasive, and easy-to-use method to quantify endothelial permeability and dysfunction in the arteries using Magnetic Resonance Imaging (MRI), named qMETRIC. This technique enables assessing vascular damage and cardiovascular risk associated with atherosclerosis in preclinical models and humans.

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Behavior

An Inescapable Cat Odor Exposure Protocol for Studying Innate and Contextual Threat Conditioning in Rats
María Rodríguez 1,2, Marco Contreras 3, Marcos Domic-Siede 4, Francisco Ceric 5, Fernando Torrealba 1
1Departamento de Fisiología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, 2Instituto de Ciencias Sociales, Universidad de O'Higgins, 3Department of Psychology, University of Arizona, Tucson, 4Escuela de Psicología, Universidad Católica del Norte, 5Laboratorio de Neurociencia Afectiva, Facultad de Psicología, Universidad del Desarrollo

Cat skin/fur odor exposure provides a reliable method to investigate neural circuits and mechanisms of defensive responses in rodents and may offer insight into mechanisms that mediate fear in humans. Here, we describe a protocol for investigating the role of the interoceptive cortex in threat responses in rats.

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Biology

Differentiation and Imaging of Brown Adipocytes from the Stromal Vascular Fraction of Interscapular Adipose Tissue from Newborn Mice
Ana-María Figueroa 1, Francisca Stolzenbach 1, Pablo Tapia 1, Víctor Cortés 1
1Department of Nutrition, Diabetes and Metabolism, Pontificia Universidad Católica de Chile

Preadipocytes are isolated from the stromal vascular fraction of interscapular brown adipose tissue from newborn mice and differentiated into cells that accumulate lipid droplets, express molecular markers, and show the mitochondrial morphology of mature brown adipocytes. These cells are further analyzed by immunofluorescence and transmission electron microscopy.

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Neuroscience

A Visuospatial Planning Task Coupled with Eye-Tracker and Electroencephalogram Systems
Marcos Domic-Siede 1, Martín Irani 2, Joaquín Valdés 3, María Rodríguez 4,5, Brice Follet 3,6, Marcela Perrone-Bertolotti 7,8, Tomás Ossandón 3
1Laboratorio de Neurociencia Cognitiva, Escuela de Psicología, Universidad Católica del Norte, 2Department of Psychology, University of Illinois Urbana-Champaign, 3Neurodynamics of Cognition Laboratory, Departamento de Psiquiatría, Pontificia Universidad Católica de Chile, 4Instituto de Ciencias de la Salud, Universidad de O’Higgins, 5Instituto de Ciencias de la Ingeniería, Universidad de O’Higgins, 6Department of Psychiatry, University Hospital of Saint-Etienne, 7Univ. Grenoble Alpes, Univ. Savoie Mont Blanc, CNRS, LPNC, 8Institut Universitaire de France (IUF)

The study of cognitive planning combining EEG and eye-tracking systems provides a multimodal approach to investigate the neural mechanisms that mediate cognitive control and goal-directed behavior in humans. Here, we describe a protocol for investigating the role of brain oscillations and eye movements in planning performance.

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Biochemistry

Optimized Methods for the Surface Immobilization of Collagens and Collagen Binding Assays
Nadia Chaher 1, Giuseppe Digilio *2, Sara Lacerda *3, René M. Botnar 1,4,5,6, Alkystis Phinikaridou 1,4
1School of Biomedical Engineering and Imaging Sciences, King’s College London, 2Dipartimento di Scienze e Innovazione Tecnologica, Università del Piemonte Orientale, 3Centre de Biophysique Moléculaire, CNRS UPR 4301, Université d'Orléans, 4BHF Centre of Research Excellence, Cardiovascular Division, King’s College London, 5Escuela de Ingeniería, Pontificia Universidad Católica de Chile, 6Instituto de Ingeniería Biológica y Médica, Pontificia Universidad Católica de Chile

This work presents an optimized protocol to reproducibly immobilize and quantify type I and III collagen onto microplates, followed by an improved in vitro binding assay protocol to study collagen-compound interactions using a time-resolved fluorescence method. The subsequent step-by-step data analysis and data interpretation are provided.

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Biology

Vertical Immobilization Method for Time-Lapse Microscopy Analysis in Filamentous Cyanobacteria
Jorge Olivares 1, Annia González 1, Derly Andrade 1,2, Mónica Vásquez 1
1Departamento de Genética Molecular y Microbiología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, 2Laboratorio de Ciencias Omicas, Facultad de Ciencias de la Salud, Universidad Espíritu Santo

We present a simple and accessible method for filamentous cyanobacterial visualization in the XY plane. A low-melting point agarose matrix was used, allowing the acquisition of images of proteins involved in the division, in a vertical orientation. Therefore, this methodology can be applied to any filamentous organism and different kinds of proteins.

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