University of South Carolina

18 articles published in JoVE

• Developmental Biology

  
  Establishing a High Throughput Epidermal Spheroid Culture System to Model Keratinocyte Stem Cell Plasticity Yvon Woappi^1,2, Geraldine Ezeka³, Justin Vercellino⁴, Sean M. Bloos⁵, Kim E. Creek⁶, Lucia Pirisi^{1 1}Department of Pathology, Microbiology and Immunology, University of South Carolina School of Medicine, ² Here we describe a protocol for the systematic cultivation of epidermal spheroids in 3D suspension culture. This protocol has wide-ranging applications for use in a variety of epithelial tissue types and for the modeling of several human diseases and conditions.

• Neuroscience

  
  A Rat Model of EcoHIV Brain Infection Hailong Li¹, Kristen A. McLaurin¹, Charles F. Mactutus¹, Rosemarie M. Booze^{1 1}Program in Behavioral Neuroscience, Department of Psychology, University of South Carolina Here, we present a protocol to establish a new rat model of active HIV infection using chimeric HIV (EcoHIV), which is critical for enhancing our understanding of HIV-1 viral reservoirs in the brain and offering a system to study HIV-associated neurocognitive disorders and associated comorbidities (i.e., drug abuse).

• Neuroscience

  
  A Hydrophobic Tissue Clearing Method for Rat Brain Tissue Kristin N. Kirchner¹, Hailong Li¹, Adam R. Denton¹, Steven B. Harrod¹, Charles F. Mactutus¹, Rosemarie M. Booze^{1 1}Program in Behavioral Neuroscience, Department of Psychology, University of South Carolina Here we present a hydrophobic tissue clearing method that allows for the viewing of target molecules as part of intact brain structures. This technique has now been validated for F344/N control and HIV-1 transgenic rats of both sexes.

• Neuroscience

  
  Ballistic Labeling of Pyramidal Neurons in Brain Slices and in Primary Cell Culture Hailong Li¹, Kristen A. McLaurin¹, Charles F. Mactutus¹, Rosemarie M. Booze^{1 1}Program in Behavioral Neuroscience, Department of Psychology, University of South Carolina We present a protocol to label and analyze pyramidal neurons, which is critical for evaluating potential morphological alterations in neurons and dendritic spines that may underlie neurochemical and behavioral abnormalities.

• Bioengineering

  
  Encapsulated Cell Technology for the Delivery of Biologics to the Mouse Eye Marwa Belhaj¹, Balasubramaniam Annamalai², Nathaniel Parsons², Andrew Shuler¹, Jay Potts¹, Bärbel Rohrer^{2,3,4 1}Department of Cell Biology, University of South Carolina, ²Department of Ophthalmology, Division of Research, Medical University of South Carolina, ³Department of Neuroscience, Division of Research, Medical University of South Carolina, ⁴Division of Research, Ralph H. Johnson VA Medical Center Presented here is a protocol for the use of alginate as a polymer in microencapsulation of immortalized cells for long-term delivery of biologics to rodent eyes.

• Neuroscience

  
  The Power of Interstimulus Interval for the Assessment of Temporal Processing in Rodents Kristen A McLaurin¹, Landhing M Moran¹, Hailong Li¹, Rosemarie M Booze¹, Charles F Mactutus^{1 1}Program in Behavioral Neuroscience, Department of Psychology, University of South Carolina Temporal processing, a preattentive process, may underlie deficits in higher-level cognitive processes, including attention, commonly observed in neurocognitive disorders. Using prepulse inhibition as an exemplar paradigm, we present a protocol for manipulating interstimulus interval (ISI) to establish the shape of the ISI function to provide an assessment of temporal processing.

• Environment

  
  Specific and Accurate Detection of the Citrus Greening Pathogen Candidatus liberibacter spp. Using Conventional PCR on Citrus Leaf Tissue Samples Huan Chen*^1,2, Ian Arthur Palmer*¹, Jian Chen^1,2, Ming Chang^1,2, Stephen L. Thompson³, Fengquan Liu², Zheng Qing Fu^{1 1}Department of Biological Sciences, University of South Carolina, ²Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, ³Department of Instructions and Teacher Education, University of South Carolina Citrus Greening is a particularly destructive disease affecting citrus crops globally. Presented here is a simple method using PCR and genomic DNA extraction of citrus leaf tissue for the accurate and precise identification of the citrus greening pathogen, Candidatus liberibacter spp.

• 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. Booze^{1 1}Program 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.

• Chemistry

  
  Characterization, Quantification and Compound-specific Isotopic Analysis of Pyrogenic Carbon Using Benzene Polycarboxylic Acids (BPCA) Daniel B. Wiedemeier¹, Susan Q. Lang², Merle Gierga³, Samuel Abiven¹, Stefano M. Bernasconi³, Gretchen L. Früh-Green³, Irka Hajdas⁴, Ulrich M. Hanke¹, Michael D. Hilf¹, Cameron P. McIntyre⁴, Maximilian P. W. Scheider¹, Rienk H. Smittenberg⁵, Lukas Wacker⁴, Guido L. B. Wiesenberg¹, Michael W. I. Schmidt^{1 1}Department of Geography, University of Zurich, ²Department of Earth and Ocean Sciences, University of South Carolina, ³Department of Earth Sciences, ETH Zurich, ⁴Laboratory of Ion Beam Physics, ETH Zurich, ⁵Department of Geological Sciences, Stockholm University We present the benzene polycarboxylic acid (BPCA) method for assessing pyrogenic carbon (PyC) in the environment. The compound-specific approach uniquely provides simultaneous information about the characteristics, quantity and isotopic composition (¹³C and ¹⁴C) of PyC.

• Developmental Biology

  
  A Novel Ex Ovo Banding Technique to Alter Intracardiac Hemodynamics in an Embryonic Chicken System Vinal Menon¹, Lorain Junor², Marwa Balhaj¹, John F. Eberth^1,3, Jay D. Potts^{1,3 1}Department of Cell Biology and Anatomy, School of Medicine, University of South Carolina, ²Instrumentation Resource Facility, School of Medicine, University of South Carolina, ³Biomedical Engineering Program, College of Engineering and Computing, University of South Carolina For the first time we present here a reproducible banding procedure to alter hemodynamics in the developing heart ex ovo. This is achieved by partially constricting the outflow tract (OFT).

• Bioengineering

  
  Three-dimensional Biomimetic Technology: Novel Biorubber Creates Defined Micro- and Macro-scale Architectures in Collagen Hydrogels Veronica Rodriguez-Rivera¹, John W. Weidner², Michael J. Yost^{1 1}Department of Surgery - Division of General Surgery, Medical University of South Carolina, ²Department of Chemical Engineering, University of South Carolina An innovative biofabrication technique was developed to engineer three-dimensional constructs that resemble the architectural features, components, and mechanical properties of in vivo tissue. This technique features a newly developed sacrificial material, BSA rubber, which transfers detailed spatial features, reproducing the in vivo architectures of a wide variety of tissues.

• Biology

  
  Using Digital Image Correlation to Characterize Local Strains on Vascular Tissue Specimens Boran Zhou¹, Suraj Ravindran², Jahid Ferdous², Addis Kidane², Michael A. Sutton², Tarek Shazly^{1,2 1}Biomedical Engineering Program, University of South Carolina, ²Department of Mechanical Engineering, University of South Carolina We describe the use of digital image correlation to characterize the local surface strain field on vascular tissue samples subjected to uniaxial tensile testing. These measurements facilitate precise quantification of the sample mechanical response and the generation of constitutive stress-strain relations.

• Biology

  
  Development of Inhibitors of Protein-protein Interactions through REPLACE: Application to the Design and Development Non-ATP Competitive CDK Inhibitors Padmavathy Nandha Premnath¹, Sandra Craig¹, Campbell McInnes^{1 1}Department of Drug Discovery and Biomedical Sciences, University of South Carolina We describe implementation of the REPLACE strategy for targeting protein-protein interactions. REPLACE is an iterative strategy involving synthetic and computational approaches for the conversion of optimized peptidic inhibitors into drug like molecules.

• Biology

  
  A Fluorescent Screening Assay for Identifying Modulators of GIRK Channels Maribel Vazquez¹, Charity A. Dunn¹, Kenneth B. Walsh^{1 1}Department 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. Blanchette^{2 1}Biomedical 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. Janicki^{1 1}Department 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 Norman^{1 1}Department 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. Welch^{2 1}Department 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.