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Universidad de Valencia

4 ARTICLES PUBLISHED IN JoVE

Developmental Biology

Stable and Efficient Genetic Modification of Cells in the Adult Mouse V-SVZ for the Analysis of Neural Stem Cell Autonomous and Non-autonomous Effects

Eva Porlan¹, Beatriz Martí-Prado^2,3, Antonella Consiglio^4,5, Isabel Fariñas^2,3

¹Cell Division and Cancer Group, Spanish National Cancer Research Centre (CNIO), ²Centro de Investigaciones Biomédicas en Red sobre Enfermedades Neurodegenerativas (CIBERNED), ³Departmento de Biologìa Celular, Universidad de Valencia, ⁴Institut de Biomedicina de la Universitat de Barcelona (IBUB), ⁵Department of Molecular and Translational Medicine, Fibroblast Reprogramming Unit, University of Brescia

Here we describe a procedure based on the use of lentiviral particles for the long-term genetic modification of neural stem cells and/or their adjacent ependymal cells in the adult ventricular-subventricular neurogenic niche which allows the separate analysis of cell autonomous and non-autonomous, niche-dependent effects on neural stem cells.

Neuroscience

Live Imaging Followed by Single Cell Tracking to Monitor Cell Biology and the Lineage Progression of Multiple Neural Populations

Rosa Gómez-Villafuertes^*1,2,3, Lucía Paniagua-Herranz^*1,2,3, Sergio Gascon^*4,5, David de Agustín-Durán^1,2,3, María de la O Ferreras^1,2,3, Juan Carlos Gil-Redondo^1,2,3, María José Queipo^1,2,3, Aida Menendez-Mendez^1,2,3, Ráquel Pérez-Sen^1,2,3, Esmerilda G. Delicado^1,2,3, Javier Gualix^1,2,3, Marcos R. Costa⁶, Timm Schroeder⁷, María Teresa Miras-Portugal^1,2,3, Felipe Ortega^1,2,3

¹Biochemistry and Molecular Biology Department, Faculty of Veterinary medicine, Complutense University, ²University Institute for Neurochemistry Research (IUIN), ³Instituto de Investigación Sanitaria del Hospital Clínico San Carlos (IdISSC), ⁴Institute of Stem Cell Research, Helmholtz Center Munich, Neuherberg/Munich, Germany Physiological Genomics, Biomedical Center, Ludwig-Maximilians University Munich, ⁵Toxicology and Pharmacology Department, Faculty of Veterinary medicine, Complutense University, ⁶Brain Institute, Federal University of Rio Grande do Norte, ⁷Department of Biosystems Science and Engineering, Eidgenössische Technische Hochschule (ETH) Zurich

A robust protocol to monitor neural populations by time-lapse video-microscopy followed by software-based post-processing is described. This method represents a powerful tool to identify biological events in a selected population during live imaging experiments.

Biology

Noninvasive Monitoring of Lesion Size in a Heterologous Mouse Model of Endometriosis

Jessica Martinez¹, Viviana Bisbal², Nerea Marin², Antonio Cano^1,3,4, Raul Gómez¹

¹Instituto de Investigación Sanitaria, ²Unidad de Animalario del Centro de Investigación Príncipe Felipe (CIPF), ³Departamento de Pediatría, Obstetricia y Ginecología, Universidad de Valencia, ⁴Servicio de Obstetricia y Ginecología, Hospital Clínico Universitario de Valencia

Here, we present a protocol for live-imaging of fluorescently labeled human endometrial fragments grafted in mice. The method allows studying the effects of drugs of choice on endometriotic lesion size through monitoring and quantification of fluorescence emitted by the fluorescent reporter on real time

Neuroscience

Double In Utero Electroporation to Target Temporally and Spatially Separated Cell Populations

Isabel Mateos-White¹, Jaime Fabra-Beser¹, David de Agustín-Durán¹, Cristina Gil-Sanz¹

¹Estructura de Recerca Interdisciplinar en Biotecnología y Biomedicina (ERI BIOTECMED), Departamento de Biología Celular, Biología Funcional y Antropología Física, Universidad de Valencia

Double in utero electroporation allows targeting cell populations that are spatially and temporally separated. This technique is useful to visualize interactions between those cell populations using fluorescent proteins in normal conditions but also after functional experiments to perturb genes of interest.