Ludwig-Maximilians University Munich

3 ARTICLES PUBLISHED IN JoVE

Neuroscience

Lineage-reprogramming of Pericyte-derived Cells of the Adult Human Brain into Induced Neurons

Marisa Karow¹, Christian Schichor², Ruth Beckervordersandforth³, Benedikt Berninger^1,4

¹Department of Physiological Genomics, Institute of Physiology, Ludwig Maximilians University Munich, ²Tumor Biology Lab, Neurosurgical Clinic, Ludwig-Maximilians University Munich, ³Institut für Biochemie, Emil-Fischer-Zentrum, Friedrich-Alexander-Universität Erlangen-Nürnberg, ⁴Institute of Physiological Chemistry and Focus Program Translational Neuroscience, Johannes Gutenberg University Mainz

Targeting brain-resident cells for direct lineage-reprogramming offers new perspectives for brain repair. Here we describe a protocol of how to prepare cultures enriched for brain-resident pericytes from the adult human cerebral cortex and convert these into induced neurons by retrovirus-mediated expression of the transcription factors Sox2 and Ascl1.

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.

Biochemistry

An Improved Method to Isolate Mitochondrial Contact Sites

Siavash Khosravi^*1,2, Johanna Frickel^*1, Max E. Harner¹

¹Department of Cell Biology, Biomedical Center, Medical Faculty, Ludwig-Maximilians University Munich, ²Cardio-Metabolic Diseases, Boehringer Ingelheim Pharma GmbH & Co. KG

Mitochondrial contact sites are protein complexes that interact with mitochondrial inner and outer membrane proteins. These sites are essential for the communication between the mitochondrial membranes and, thus, between the cytosol and the mitochondrial matrix. Here, we describe a method to identify candidates qualifying for this specific class of proteins.