University Hospital Wuerzburg 6 articles published in JoVE Neuroscience Laser Microdissection-Based Protocol for the LC-MS/MS Analysis of the Proteomic Profile of Neuromelanin Granules Maximilian Wulf1,2, Katalin Barkovits-Boeddinghaus1,2, Paula Sommer1,2, Karin Schork1,2, Martin Eisenacher1,2, Peter Riederer3,4, Manfred Gerlach5, Steffen Kösters1,2, Britta Eggers*1,2, Katrin Marcus*1,2 1Medizinisches Proteom-Center, Medical Faculty, Ruhr-University Bochum, 2Medical Proteome Analysis, Center for Proteindiagnostics (PRODI), Ruhr-University Bochum, 3Center of Mental Health; Clinic and Policlinic for Psychiatry, Psychosomatics and Psychotherapy, University Hospital Wuerzburg, 4Psychiatry Department of Clinical Research, University of Southern Denmark Odense University Hospital, 5Center of Mental Health, Department of Child and Adolescent Psychiatry, Psychosomatics and Psychotherapy, University Hospital of Wuerzburg, University of Wuerzburg A robust protocol is presented here for isolating neuromelanin granules from human post-mortem substantia nigra pars compacta tissue via laser microdissection. This revised and optimized protocol massively minimizes the required time for sample collection, reduces the required sample amount, and enhances the identification and quantification of proteins by LC-MS/MS analysis. Bioengineering Recombinant Collagen I Peptide Microcarriers for Cell Expansion and Their Potential Use As Cell Delivery System in a Bioreactor Model Melva Suarez Muñoz1, Davide Confalonieri1, Heike Walles1,2, Elisabeth M. W. M. van Dongen3, Gudrun Dandekar1,2 1Department Tissue Engineering and Regenerative Medicine, University Hospital Wuerzburg, 2Translational Center Regenerative Therapies (TLC-RT), Fraunhofer Institute for Silicate Research ISC, 3Fujifilm Manufacturing Europe B.V. We propose a cell expansion protocol on macroporous microcarriers and their use as delivery system in a perfusion bioreactor to seed a decellularized tissue matrix. We also include different techniques to determine cell proliferation and viability of cells cultured on microcarriers. Furthermore, we demonstrate functionality of cells after bioreactor cultures. Neuroscience Stereological Estimation of Dopaminergic Neuron Number in the Mouse Substantia Nigra Using the Optical Fractionator and Standard Microscopy Equipment Chi Wang Ip1, David Cheong1, Jens Volkmann1 1Department of Neurology, University Hospital of Würzburg This work presents a step-by-step protocol for the unbiased stereological estimation of dopaminergic neuronal cell numbers in the mouse substantia nigra using standard microscopy equipment (i.e., a light microscope, a motorized object table (x, y, z plane), and public domain software for digital image analysis. Behavior A Novel Approach to Assess Motor Outcome of Deep Brain Stimulation Effects in the Hemiparkinsonian Rat: Staircase and Cylinder Test Marta Rattka1, Felix Fluri1, Miloš Krstić1, Esther Asan2, Jens Volkmann1 1Department of Neurology, University Hospital Wuerzburg, 2Institute of Anatomy and Cell Biology, University Wuerzburg Deep brain stimulation (DBS) is an effective treatment option for Parkinson's disease. We established a study design to screen novel stimulation paradigms in rats. The protocol describes the use of the staircase test and cylinder test for motor outcome assessment in DBS treated hemiparkinsonian rats. Bioengineering A Combined 3D Tissue Engineered In Vitro/In Silico Lung Tumor Model for Predicting Drug Effectiveness in Specific Mutational Backgrounds Claudia Göttlich*1, Lena C. Müller*1, Meik Kunz*3, Franziska Schmitt1, Heike Walles1,4, Thorsten Walles2, Thomas Dandekar3, Gudrun Dandekar1,4, Sarah L. Nietzer1 1Department of Tissue Engineering and Regenerative Medicine (TERM), University Hospital Wuerzburg, 2Department of Cardiothoracic Surgery, University Hospital Wuerzburg, 3Department of Bioinformatics, University Wuerzburg, 4Translational Center Wuerzburg, Fraunhofer Institute Interfacial Engineering and Biotechnology IGB We present a three-dimensional (3D) lung cancer model based on a biological collagen scaffold to study sensitivity towards non-small-cell-lung-cancer-(NSCLC)-targeted therapies. We demonstrate different read-out techniques to determine the proliferation index, apoptosis and epithelial-mesenchymal transition (EMT) status. Collected data are integrated into an in silico model for prediction of drug sensitivity. Neuroscience Microelectrode Guided Implantation of Electrodes into the Subthalamic Nucleus of Rats for Long-term Deep Brain Stimulation Felix Fluri1, Micheal Bieber1, Jens Volkmann1, Christoph Kleinschnitz1 1Department of Neurology, University Hospital Wuerzburg A method for implanting electrodes into the subthalamic nucleus (STN) of rats is described. Better localization of the STN was achieved by using a microrecording system. Furthermore, a stimulation set-up is presented that is characterized by long-lasting connections between the head of the animal and the stimulator.