University of California, San Diego

14 articles published in JoVE

A High-throughput Method for Measurement of Glomerular Filtration Rate in Conscious Mice

JoVE 50330 5/10/2013

¹Division of Nephrology-Hypertension, Department of Medicine, University of California, San Diego, ²San Diego VA Healthcare System

Measurement of glomerular filtration rate (GFR) is the gold standard for kidney function assessment. Here we describe a high-throughput method which allows the determination of GFR in conscious mice by using a single bolus injection, determination of fluorescein isothiocyanate (FITC)-inulin in plasma and calculation of GFR by a two-phase exponential decay model.

Density Gradient Multilayered Polymerization (DGMP): A Novel Technique for Creating Multi-compartment, Customizable Scaffolds for Tissue Engineering

JoVE 50018 2/12/2013

Shivanjali Joshi-Barr¹, Jerome V. Karpiak², Yogesh Ner¹, Jessica H. Wen³, Adam J. Engler³, Adah Almutairi^1,2

¹Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego, ²Biomedical Sciences Program, University of California, San Diego, ³Department of Bioengineering, University of California, San Diego

Here we describe a unique strategy for creating biocompatible, layered matrices with continuous interfaces between distinct layers for tissue engineering. Such a scaffold could provide an ideal customizable environment to modulate cell behavior by various biological, chemical or mechanical cues

Using Unfixed, Frozen Tissues to Study Natural Mucin Distribution

JoVE 3928 9/21/2012

Miriam Cohen¹, Nissi M. Varki¹, Mark D. Jankowski², Pascal Gagneux¹

¹Department of Cellular and Molecular Medicine, University of California, San Diego, ²Biosecurity and Public Health, Los Alamos National Laboratory

Unfixed frozen tissue samples embedded in Optimal Cutting Temperature medium (OCT) can be used to study natural distribution and glycosylation of secreted mucus. In this approach tissue processing is minimal and the natural presentation of glycolipids, mucins and glycan-epitopes is preserved. Tissue sections can be analyzed by immunohistochemistry using fluorescence or chromogenic detection.

Sampling Human Indigenous Saliva Peptidome Using a Lollipop-Like Ultrafiltration Probe: Simplify and Enhance Peptide Detection for Clinical Mass Spectrometry

JoVE 4108 8/07/2012

Wenhong Zhu¹, Richard L. Gallo^2,3, Chun-Ming Huang^2,3,4

¹Sanford-Burnham Medical Research Institute, ²Division of Dermatology, University of California, San Diego, ³VA San Diego Healthcare Center, ⁴Moores Cancer Center, University of California, San Diego

Considering saliva sampling for future clinical application, a lollipop-like ultrafiltration (LLUF) probe was fabricated to fit in the human oral cavity. Direct analysis of undigested saliva by NanoLC-LTQ mass spectrometry demonstrated the ability of LLUF probes to remove large proteins and high abundance proteins, and make low-abundant peptides more detectable.

Detection and Genogrouping of Noroviruses from Children's Stools By Taqman One-step RT-PCR

JoVE 3232 7/22/2012

Sonia Apaza¹, Susan Espetia¹, Robert H. Gilman^1,2, Sonia Montenegro³, Susana Pineda³, Fanny Herhold¹, Romeo Pomari¹, Margaret Kosek², Nancy Vu¹, Mayuko Saito^1,2,4

¹Laboratorio de Investigación y Desarrollo (LID), Universidad Peruana Cayetano Heredia, ²Bloomberg School of Public Health, Johns Hopkins University, ³Laboratorio de Diagnostico Molecular, Facultad de Medicina, University of Concepcion,Chile, ⁴University of California San Diego School of Medicine

A One-Step RT-PCR assay for detection and genogroup identification of Norovirus isolates from children’s stools, that utilizes primers and TaqMan probes specific to the open reading frame 1 (ORF1)-ORF2 junction region, the most conserved region of the Norovirus genome is described. A non-commercial, cost-effective RNA extraction method is detailed.

A Polished and Reinforced Thinned-skull Window for Long-term Imaging of the Mouse Brain

JoVE 3742 3/07/2012

Andy Y. Shih¹, Celine Mateo¹, Patrick J. Drew^2,3, Philbert S. Tsai¹, David Kleinfeld^1,4

¹Department of Physics, University of California, San Diego, ²Department of Engineering Science and Mechanics, Pennsylvania State University, ³Department of Neurosurgery, Pennsylvania State University, ⁴Section of Neurobiology, University of California, San Diego

We present a method to form an imaging window in the mouse skull that spans millimeters and is stable for months without inflammation of the brain. This method is well suited for longitudinal studies of blood flow, cellular dynamics, and cell/vascular structure using two-photon microscopy.

Targeting Olfactory Bulb Neurons Using Combined In Vivo Electroporation and Gal4-Based Enhancer Trap Zebrafish Lines

JoVE 2964 8/15/2011

Kenric J. Hoegler¹, Martin Distel², Reinhard W. Köster³, John H. Horne¹

¹Department of Biology, Pace University, ²Cellular and Molecular Medicine, University of California, San Diego, ³Division of Cell Biology and Cell Physiology, Zoological Institute, Braunschweig University of Technology

The temporal and spatial resolution of genetic manipulations determines the spectrum of biological phenomena that they can perturb. Here we use temporally and spatially discrete in vivo electroporation, combined with transgenic lines of zebrafish, to induce expression of a GFP transgene specifically in neurons of the developing olfactory bulb.

Magnetic Resonance Imaging Quantification of Pulmonary Perfusion using Calibrated Arterial Spin Labeling

JoVE 2712 5/30/2011

Tatsuya J. Arai^1,2, G. Kim Prisk^1,3, Sebastiaan Holverda¹, Rui Carlos Sá¹, Rebecca J. Theilmann³, A. Cortney Henderson¹, Matthew V. Cronin³, Richard B. Buxton³, Susan R. Hopkins^1,3

¹Medicine, University of California San Diego - UCSD, ²Bioengineering, University of California San Diego - UCSD, ³Radiology, University of California San Diego - UCSD

A MR imaging method to study the distribution of pulmonary blood flow under a variety of physiological conditions, in this case exposure to three different inspired oxygen concentrations: hypoxia, normoxia, and hyperoxia, is described. This technique utilizes human pulmonary physiology research techniques in an MR scanning environment.

Thermodynamics of Membrane Protein Folding Measured by Fluorescence Spectroscopy

JoVE 2669 4/28/2011

Diana E. Schlamadinger, Judy E. Kim

Chemistry and Biochemistry, University of California San Diego - UCSD

This video article details the experimental procedure for obtaining the Gibbs free energy of membrane protein folding by tryptophan fluorescence.

Fabrication of Biologically Derived Injectable Materials for Myocardial Tissue Engineering

JoVE 2109 12/20/2010

Sonya Seif-Naraghi, Jennifer Singelyn, Jessica DeQuach, Pamela Schup-Magoffin, Karen Christman

University of California, San Diego

Methods for preparing an injectable matrix gel from decellularized tissue and injecting it into rat myocardium in vivo are described.

Intravital Microscopy of the Mouse Brain Microcirculation using a Closed Cranial Window

JoVE 2184 11/18/2010

Pedro Cabrales¹, Leonardo J. M. Carvalho²

¹Bioengineering, University of California, San Diego, ²La Jolla Bioengineering Institute

Intravital microscopy to follow temporal and spatial hemodynamic and inflammatory events in the pial microcirculation.

A Simple Composite Phenotype Scoring System for Evaluating Mouse Models of Cerebellar Ataxia

JoVE 1787 5/21/2010

Stephan J. Guyenet¹, Stephanie A. Furrer², Vincent M. Damian¹, Travis D. Baughan², Albert R. La Spada*³, Gwenn A. Garden*²

¹Department of Biochemistry, University of Washington, ²Department of Neurology, University of Washington, ³Division of Genetics, Departments of Pediatrics and Cellular and Molecular Medicine, and the Institute for Genomic Medicine, University of California, San Diego - Rady Children’s Hospital

We describe a protocol for the rapid and sensitive quantification of disease severity in mouse models of cerebellar ataxia. Measures include hind limb clasping, ledge test, gait and kyphosis. This protocol effectively discriminates between affected and non-affected individuals, and detects the progression of affected individuals over time.

Spinal Cord Electrophysiology

JoVE 1660 1/18/2010

Allyn Meyer¹, Benjamin W. Gallarda^1,2, Samuel Pfaff¹, William Alaynick¹

¹The Salk Institute for Biological Studies, Howard Hughes Medical Institute and Gene Expression Laboratory, ²Biology Graduate Program, University of California San Diego - UCSD

A demonstration of the isolation of neonatal mouse spinal cord for electrophysiologic studies.

Localized RNAi and Ectopic Gene Expression in the Medicinal Leech

JoVE 697 4/17/2008

Orit Shefi¹, Claire Simonnet², Alex Groisman², Eduardo R Macagno¹

¹Division of Biological Sciences, University of California San Diego - UCSD, ²Department of Physics, University of California San Diego - UCSD

In this video, we show a procedure for an accurate biolistic delivery of reagents into live tissue with a novel miniature gene gun. We are knocking down the expression of the axon guidance molecule Netrin in leech embryos by delivering molecules of dsRNA into the ventral body wall and ganglia of single segments.