University of South Carolina View Institution's Website 6 articles published in JoVE Neuroscience Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology Hailong Li1, Jessica M. Illenberger1, Kristen A. McLaurin1, Charles F. Mactutus1, Rosemarie M. Booze1 1Program in Behavioral Neuroscience, Department of Psychology, University of South Carolina Identification of dopamine D1-alpha receptor in the nucleus accumbens is critical for clarifying D1 receptor dysfunction during a central nervous system disease. We performed a novel RNA in situ hybridization assay to visualize single RNA molecules in a specific brain area. Biology A Fluorescent Screening Assay for Identifying Modulators of GIRK Channels Maribel Vazquez1, Charity A. Dunn1, Kenneth B. Walsh1 1Department of Pharmacology, Physiology & Neuroscience, University of South Carolina, School of Medicine A real-time screening procedure for identifying drugs that interact with G protein-gated inward rectifier K+ (GIRK) channels is described. The assay utilizes membrane potential-sensitive fluorescent dyes to measure GIRK channel activity. This technique is adaptable for use on a number of cell lines. Bioengineering Tracking Hypoxic Signaling within Encapsulated Cell Aggregates Matthew L. Skiles1, Suchit Sahai1, James O. Blanchette2 1Biomedical Engineering Program, University of South Carolina, 2Chemical Engineering Department, University of South Carolina A method for photo-encapsulation of cells in a crosslinked PEG hydrogel is described. Hypoxic signaling within encapsulated murine insulinoma (MIN6) aggregates is tracked using a fluorescent marker system. This system allows serial examination of cells within a hydrogel scaffold and correlation of hypoxic signaling with changes in cell phenotype. Immunology and Infection Isolation of Functional Cardiac Immune Cells Jennifer L. McLarty1, Giselle C. Meléndez1, William J. Spencer1, Scott P. Levick1, Gregory L. Brower1, Joseph S. Janicki1 1Department of Cell Biology and Anatomy, University of South Carolina- School of Medicine This method for isolating functional immune cells from the heart provides an alternative to the conventional methods of collagenase digestion, which causes unwanted immune cell activation, resulting in a decreased responsiveness of these cells. Our method of isolation yields functional cardiac immune cells by avoiding problems associated with enzymatic digestion. Biology Extraction of High Molecular Weight DNA from Microbial Mats Benjamin S. Bey1, Erin B. Fichot1, R. Sean Norman1 1Department of Environmental Health Sciences, Arnold School of Public Health, University of South Carolina We provide an improved protocol for extracting high molecular weight DNA from hypersaline microbial mats. Microbial cells are separated from the mat matrix prior to DNA extraction and purification. This enhances the concentrations, quality, and size of the DNA. The protocol may be used for other refractory samples. Biology Recording Multicellular Behavior in Myxococcus xanthus Biofilms using Time-lapse Microcinematography Rion G. Taylor1, Roy D. Welch2 1Department of Environmental Health Sciences, University of South Carolina (USC), 2Department of Biology, Syracuse University To study Myxococcus xanthus swarm behavior, we have designed a time-lapse microcinematography protocol that can be modified for different assays. It employs standard growth conditions adapted for microscopy, and yields reproducible results by the use of inexpensive, reusable silicone gaskets. We have used this method to quantify multicellular chemotaxis.
