Vanderbilt University School of Medicine 9 articles published in JoVE Biology Human Pseudoislet System for Synchronous Assessment of Fluorescent Biosensor Dynamics and Hormone Secretory Profiles Tiffany M. Richardson*1, Yasminye D. Pettway*1, John T. Walker1, Heather A. Nelson2, Matthew Ishahak3, Gregory Poffenberger2, Radhika Aramandla2, Conrad Reihsmann2, Ashutosh Agarwal3, Alvin C. Powers1,2,4, Marcela Brissova2 1Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, 2Division of Diabetes, Endocrinology and Metabolism, Department of Medicine, Vanderbilt University Medical Center, 3Department of Biomedical Engineering, University of Miami, 4VA Tennessee Valley Healthcare System This protocol describes a method for the synchronous acquisition and co-registration of intracellular signaling events and the secretion of insulin and glucagon by primary human pseudoislets using the adenoviral delivery of a cyclic adenosine monophosphate (cAMP) biosensor, a cAMP difference detector in situ (cADDis), and a microperifusion system. Biology Real-Time Measurements of Calcium and Contractility Parameters in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes Rafeeh Dinani1,2, Emmy Manders1,2,3, Michiel Helmes1,2,3, Lili Wang4, Bjorn C. Knollmann4, Diederik W. D. Kuster1,2, Jolanda van der Velden1,2 1Division of Physiology, Amsterdam UMC, Vrije University, 2Heart Failure & Arrhythmias, Amsterdam Cardiovascular Sciences, 3CytoCypher BV, 4Division of Clinical Pharmacology, Vanderbilt School of Medicine Here, an established method is described to perform contractility and calcium measurements in human induced pluripotent stem cell-derived cardiomyocytes using an optics-based platform. This platform enables researchers to study the effect of mutations and the response to various stimuli in a rapid and reproducible manner. Biology Optimization of Transesophageal Atrial Pacing to Assess Atrial Fibrillation Susceptibility in Mice Matthew B. Murphy1, Kyungsoo Kim1, Prince J. Kannankeril1, Katherine T. Murray1 1Departments of Medicine, Pediatrics and Pharmacology, Vanderbilt University School of Medicine The present protocol describes the optimization of experimental parameters when using transesophageal atrial pacing to assess atrial fibrillation susceptibility in mice. Medicine Blind Endotracheal Intubation in Neonatal Rabbits Sergio Benito1, Stephanie Hadley2, Marta Camprubí-Camprubí3, Joan Sanchez-de-Toledo4,5 1Department of Pediatric Critical Care, Hospital Sant Joan de Déu, Institut de Recerca Sant Joan de Déu, 2Vanderbilt University School of Medicine, 3Neonatology Department, Hospital Sant Joan de Déu, Institut de Recerca Sant Joan de Déu, Universidad de Barcelona, 4Department of Pediatric Cardiology, Hospital Sant Joan de Déu, Institut de Recerca Sant Joan de Déu, 5Department of Critical Care Medicine, University of Pittsburgh We describe a technique of endotracheal intubation in newborn rabbits after esophageal catheterization with a gastric tube. Cancer Research Perfusion and Inflation of the Mouse Lung for Tumor Histology Mackenzie L. Davenport1, Taylor P. Sherrill2, Timothy S. Blackwell2, Mick D. Edmonds1 1Department of Genetics, University of Alabama at Birmingham, 2Department of Medicine, Division of Allergy, Pulmonary, and Critical Care Medicine, Vanderbilt University School of Medicine The purpose of this method is to present a simple and efficient method for the perfusion, inflation, and fixation of mouse lungs for the examination of lung tumor pathology and evaluation of metastases to the lung. Biochemistry Isolation of High-density Lipoproteins for Non-coding Small RNA Quantification Danielle L. Michell*1, Ryan M. Allen*1, Stuart R. Landstreet1, Shilin Zhao2, Cynthia L. Toth1, Quanhu Sheng3, Kasey C. Vickers1,2 1Department of Medicine, Vanderbilt University School of Medicine, 2Center for Quantitative Sciences, Vanderbilt University School of Medicine, 3Department of Cancer Biology, Vanderbilt University School of Medicine This protocol describes the isolation and quantification of high-density lipoprotein small RNAs. Immunology and Infection High Resolution Electron Microscopy of the Helicobacter pylori Cag Type IV Secretion System Pili Produced in Varying Conditions of Iron Availability Kathryn Patricia Haley1, Eric Joshua Blanz1, Jennifer Angeline Gaddy1,2 1Department of Medicine - Division of Infectious Diseases, Vanderbilt University School of Medicine, 2Tennessee Valley Healthcare Systems, U. S. Dept. of Veterans Affairs Here we describe a method to visualize the oncogenic bacterial organelle known as the Cag Type IV Secretion System (Cag-T4SS). We find that the Cag-T4SS is differentially produced on the surface of H. pylori in response to varying conditions of iron availability. Biology Reconstitution Of β-catenin Degradation In Xenopus Egg Extract Tony W. Chen*1, Matthew R. Broadus*1, Stacey S. Huppert2, Ethan Lee1,3 1Department of Cell and Developmental Biology and Program in Developmental Biology, Vanderbilt University Medical Center, 2 A method is described for analyzing protein degradation using radiolabeled and luciferase-fusion proteins in Xenopus egg extract and its adaptation for high-throughput screening for small molecule modulators of protein degradation. Bioengineering A Step Beyond BRET: Fluorescence by Unbound Excitation from Luminescence (FUEL) Joseph Dragavon1, Carolyn Sinow2, Alexandra D. Holland1, Abdessalem Rekiki1, Ioanna Theodorou3, Chelsea Samson4, Samantha Blazquez1, Kelly L. Rogers5, Régis Tournebize1,6,7, Spencer L. Shorte1 1Plate-Forme d'Imagerie Dynamique, Imagopole, Institut Pasteur, 2Department of Radiation Oncology, Stanford School of Medicine, 3 Expanding the foundation and applicability of Fluorescence by Unbound Excitation from Luminescence (FUEL) by surveying the relevant principles and demonstrating its compatibility with a multitude of fluorophores and antibody-targeted conditions.