The University of North Carolina at Chapel Hill 6 articles published in JoVE Neuroscience Whole-Brain Single-Cell Imaging and Analysis of Intact Neonatal Mouse Brains Using MRI, Tissue Clearing, and Light-Sheet Microscopy Felix A. Kyere*1,2, Ian Curtin*1,2, Oleh Krupa1,2, Carolyn M. McCormick1,2, Mustafa Dere3, Sarah Khan3,7, Minjeong Kim7, Tzu-Wen Winnie Wang4,5,6, Qiuhong He4,5,6, Guorong Wu3, Yen-Yu Ian Shih4,5,6, Jason L. Stein1,2 1UNC Neuroscience Center, University of North Carolina, Chapel Hill, 2Department of Genetics, University of North Carolina, Chapel Hill, 3Department of Psychiatry, University of North Carolina, Chapel Hill, 4Center for Animal Magnetic Resonance Imaging, The University of North Carolina at Chapel Hill, 5Biomedical Research Imaging Center, The University of North Carolina at Chapel Hill, 6Department of Neurology, The University of North Carolina at Chapel Hill, 7Department of Computer Science, The University of North Carolina at Greensboro This protocol describes methods for conducting magnetic resonance imaging, clearing, and immunolabeling of intact mouse brains using iDISCO+, followed by a detailed description of imaging using light-sheet microscopy, and downstream analyses using NuMorph. Medicine Rapid Viscoelastic Characterization of Airway Mucus Using a Benchtop Rheometer Jason A. Wykoff1, Kendall M. Shaffer1, Kenza C. Araba1, Matthew R. Markovetz1, Jérémy Patarin2, Matthieu Robert de Saint Vincent2, Scott H. Donaldson1,3, Camille Ehre1,4 1Marsico Lung Institute / CF Center, The University of North Carolina at Chapel Hill, 2Rheonova, 3Department of Pulmonary and Critical Care Medicine, The University of North Carolina at Chapel Hill, 4Department of Pediatric, Pediatric Pulmonology Division, The University of North Carolina at Chapel Hill The viscoelastic properties of mucus play a critical role in mucociliary clearance. However, traditional mucus rheological techniques require complex and time-consuming approaches. This study provides a detailed protocol for the use of a benchtop rheometer that can rapidly and reliably perform viscoelastic measurements. Behavior Behavioral Assessment of Hearing in 2 to 4 Year-old Children: A Two-interval, Observer-based Procedure Using Conditioned Play-based Responses Angela Yarnell Bonino1,2, Lori J. Leibold3 1Department of Allied Health Sciences, The University of North Carolina at Chapel Hill, 2Department of Speech, Language, and Hearing Sciences, University of Colorado Boulder, 3Center for Hearing Research, Boys Town National Research Hospital This paper describes a procedure for measuring hearing sensitivity in 2 to 4 year-old children. Children are trained to perform play-based responses when they hear a target signal. Thresholds are then estimated in a two-interval, two-alternative forced-choice paradigm, based on observations of the child's behaviors. Neuroscience Utilizing Combined Methodologies to Define the Role of Plasma Membrane Delivery During Axon Branching and Neuronal Morphogenesis Cortney C. Winkle1, Christopher C. Hanlin2, Stephanie L. Gupton1,2 1Curriculum in Neurobiology, The University of North Carolina at Chapel Hill, 2Department of Cell Biology and Physiology, The University of North Carolina at Chapel Hill Light microscopy techniques coupled with biochemical assays elucidate the involvement of SNARE-mediated exocytosis in netrin-dependent axon branching. This combination of techniques permits identification of molecular mechanisms controlling axon branching and cell shape change. Biology Fundamental Technical Elements of Freeze-fracture/Freeze-etch in Biological Electron Microscopy Johnny L. Carson1 1Department of Pediatrics, Center for Environmental Medicine, Asthma, and Lung Biology, The University of North Carolina at Chapel Hill Basic techniques and refinements of freeze-fracture processing of biological specimens and nanomaterials for examination by transmission electron microscopy are described. This technique is a preferred method for revealing ultrastructural features and specializations of biological membranes and for obtaining ultrastructural level dimensional and spatial data in materials sciences and nanotechnology products. Biology Culturing of Human Nasal Epithelial Cells at the Air Liquid Interface Loretta Müller1, Luisa E. Brighton1, Johnny L. Carson1,2, William A. Fischer II1,3, Ilona Jaspers1,2,4 1Center for Environmental Medicine, Asthma, and Lung Biology, The University of North Carolina at Chapel Hill, 2Department of Pediatrics, The University of North Carolina at Chapel Hill, 3Pulmonary Diseases and Critical Care, The University of North Carolina at Chapel Hill, 4Curriculum in Toxicology, The University of North Carolina at Chapel Hill Nasal epithelial cells, obtained through superficial scrape biopsy of human volunteers, are expanded and transferred onto tissue culture inserts. Upon reaching confluency, cells are grown at air liquid interface, yielding cultures of ciliated and non-ciliated cells. Differentiated nasal epithelial cell cultures provide viable experimental models for studying the respiratory mucosa.
