Universidad Nacional Autonoma de Mexico 12 articles published in JoVE Medicine Lumican Extraction from Amniotic Membrane and Determination of its Storage Temperature Luis Haro-Morlett*1, Fatima Sofia Magaña-Guerrero*1, Beatriz Buentello Volante1, Yonathan Garfias1,2 1Instituto de Oftalmologia Fundacion Conde de Valenciana IAP, 2Biochemistry Department, Medicine Faculty, Universidad Nacional Autónoma de Mexico The present protocol describes the extraction of lumican from the amniotic membrane (AM) and their storing conditions as AM extract (AME) at -20 °C, 4 °C, and room temperature (RT) for 6, 12, 20, and 32 days to quantify its proteins and lumican concentration. Biochemistry Identification of Hemolytic and Phospholipase Activity in Crude Extracts from Sea Anemones by Straightforward Bioassays Santos Ramírez-Carreto1, Sandra I. Salazar-García1, Giovanni Macías Martínez1, Claudia Rodríguez-Almazán1,2 1Instituto de Biotecnología, Universidad Nacional Autónoma de México, 2Departamento de Micro y Nanotecnologías, Instituto de Ciencias Aplicadas y Tecnología, Universidad Nacional Autónoma de México Here, we describe a protocol to obtain crude venom extract from sea anemone and detect its hemolytic and phospholipase activity. Developmental Biology Analysis of Cell Differentiation, Morphogenesis, and Patterning During Chicken Embryogenesis Using the Soaked-Bead Assay Jessica Cristina Marín-Llera1, Jesús Chimal-Monroy1 1Departamento de Medicina Genómica y Toxicología Ambiental, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México The soaked bead assay involves targeted delivery of test reagent at any developmental time point to study the regulation of cell differentiation and morphogenesis. A detailed protocol, applicable to any experimental animal model, for preparing three different types of soaked beads and implanting these in the interdigit of a chicken embryo is presented. Genetics In-Nucleus Hi-C in Drosophila Cells Ayerim Esquivel-López1, Rodrigo Arzate-Mejía1, Rosario Pérez-Molina1, Mayra Furlan-Magaril1 1Departamento de Genética Molecular, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México The genome is organized in the nuclear space into different structures that can be revealed through chromosome conformation capture technologies. The in-nucleus Hi-C method provides a genome-wide collection of chromatin interactions in Drosophila cell lines, which generates contact maps that can be explored at megabase resolution at restriction fragment level. Biology Immunofluorescent Staining for Visualization of Heterochromatin Associated Proteins in Drosophila Salivary Glands Silvia Meyer-Nava1, Mario Zurita1, Viviana Valadez-Graham1 1Instituto de Biotecnología, Departamento de Genética del Desarrollo y Fisiología Molecular, Universidad Nacional Autónoma de México This protocol aims to visualize heterochromatin aggregates in Drosophila polytene cells. Neuroscience Unraveling the Role of Discrete Areas of the Rat Brain in the Regulation of Ovulation through Reversible Inactivation by Tetrodotoxin Microinjections Carlos-Camilo Silva*1,2, Montserrat Bolaños-Hurtado*1, Cinthia Juárez-Tapia4, Angélica Flores1,2, Isabel Arrieta-Cruz3,5, María-Esther Cruz3, Roberto Domínguez1,2,3 1Chronobiology of Reproduction Research Lab, Biology of Reproduction Research Unit, Facultad de Estudios Superiores Zaragoza, Universidad Nacional Autónoma de México, 2Developmental Biology Lab, Biology of Reproduction Research Unit, Facultad de Estudios Superiores Zaragoza, Universidad Nacional Autónoma de México, 3Neuroendocrinology Lab, Biology of Reproduction Research Unit, Facultad de Estudios Superiores Zaragoza, Universidad Nacional Autónoma de México, 4Facultad de Ciencias, Universidad Nacional Autónoma de México, 5Department of Basic Research, Instituto Nacional de Geriatria, Ministry of Health This protocol describes the construction of a low-cost microinjection system, its stereotaxic implantation into deep-brain structures and the procedure for timed microinjections of tetrodotoxin in awake and unrestrained rats. The goal is to reveal the participation of hypothalamic structures in the regulation of ovulation by inhibiting their neural activity. Neuroscience Culture of Neurospheres Derived from the Neurogenic Niches in Adult Prairie Voles Daniela Ávila-González1, Larry J. Young2, Francisco Camacho1, Raúl G. Paredes1,3, Néstor F. Díaz4, Wendy Portillo1 1Departamento de Neurobiología Conductual y Cognitiva, Instituto de Neurobiología, Universidad Nacional Autónoma de México, 2Silvio O. Conte Center for Oxytocin and Social Cognition, Center for Translational Social Neuroscience, Department of Psychiatry and Behavioral Sciences, Yerkes National Primate Research Center, Emory University, 3Escuela Nacional de Estudios Superiores Juriquilla, Universidad Nacional Autónoma de México, 4Departamento de Fisiología y Desarrollo Celular, Instituto Nacional de Perinatología We established the conditions to culture neural progenitor cells from the subventricular zone and dentate gyrus of the adult brain of prairie voles, as a complementary in vitro study, to analyze the sex-dependent differences between neurogenic niches that could be part of functional plastic changes associated with social behaviors. Neuroscience Desensitization and Recovery of Crayfish Photoreceptors Upon Delivery of a Light Stimulus Carolina Barriga-Montoya1, Araceli de la O-Martínez1, Arturo Picones2, Arturo Hernández-Cruz2, Beatriz Fuentes-Pardo1, Froylán Gómez-Lagunas1 1Departamento de Fisiología, Facultad de Medicina, Universidad Nacional Autónoma de México, 2Laboratorio Nacional de Canalopatías, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México A protocol for the study of desensitization and sensitivity recovery of crayfish photoreceptors as a function of circadian time is presented. Neuroscience Simple Surgical Induction of Conductive Hearing Loss with Verification Using Otoscope Visualization and Behavioral Clap Startle Response in Rat Francis A. M. Manno*1, Sinai H. C. Manno*2,3,4, Victor Ma5, Fernando A. Barrios1, William C Cho5, Shuk Han Cheng2,3,6, Condon Lau4 1Instituto de Neurobiología, Universidad Nacional Autónoma de México, México, 2State Key Laboratory of Marine Pollution (SKLMP), City University of Hong Kong, Hong Kong, 3Department of Biomedical Sciences, College of Veterinary Medicine and Life Sciences, City University of Hong Kong, Hong Kong, 4Department of Physics, City University of Hong Kong, Hong Kong, 5Department of Clinical Oncology, Queen Elizabeth Hospital, Hong Kong, 6Department of Materials Science and Engineering, City University of Hong Kong, Hong Kong Here, we present a protocol to establish a replicable conductive hearing loss induction via surgical tympanic membrane puncture and verification by otoscope visualization and behavioral assessment by clap startle. Chemistry A Facile Synthetic Method to Obtain Bismuth Oxyiodide Microspheres Highly Functional for the Photocatalytic Processes of Water Depuration Juan C. Durán-Álvarez1, Carolina Martínez1, Adriana C. Mera2, Raquel Del Angel1, Nini J. Gutiérrez-Moreno1, Rodolfo Zanella1 1Instituto de Ciencias Aplicadas y Tecnología, Universidad Nacional Autónoma de México, 2Instituto de Investigación Multidisciplinario en Ciencia y Tecnología, Universidad de La Serena This article describes a synthetic method to obtain bismuth oxyiodide microspheres, which are highly functional to perform the photocatalytic removal of organic pollutants, such as ciprofloxacin, in water under UV-A/visible light irradiation. Genetics Promoter Capture Hi-C: High-resolution, Genome-wide Profiling of Promoter Interactions Stefan Schoenfelder*1, Biola-Maria Javierre*1,2, Mayra Furlan-Magaril1,3, Steven W. Wingett1,4, Peter Fraser1,5 1Nuclear Dynamics Programme, The Babraham Institute, Babraham Research Campus, 2IJC Building, Campus ICO-Germans Trias i Pujol, Josep Carreras Leukemia Research Institute, 3Departamento de Genética Molecular, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, 4Bioinformatics Group, The Babraham Institute, Babraham Research Campus, 5Department of Biological Science, Florida State University DNA regulatory elements, such as enhancers, control gene expression by physically contacting target gene promoters, often through long-range chromosomal interactions spanning large genomic distances. Promoter Capture Hi-C (PCHi-C) identifies significant interactions between promoters and distal regions, enabling the assignment of potential regulatory sequences to their target genes. Biochemistry Lipid Index Determination by Liquid Fluorescence Recovery in the Fungal Pathogen Ustilago Maydis Lucero Romero-Aguilar*1,2, Mónica Montero-Lomeli*3, Juan Pablo Pardo2, Guadalupe Guerra- Sánchez1 1Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, 2Facultad de Medicina, Universidad Nacional Autónoma de México, 3Instituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro Here, we describe a protocol to obtain the lipid droplet index (LD index) to study the dynamics of triacylglycerols in cells cultured in high-throughput experiments. The LD index assay is an easy and reliable method that uses BODIPY 493/503. This assay does not need dispendious lipid extraction or microscopy analysis.
