University of California San Diego - UCSD

8 ARTICLES PUBLISHED IN JoVE

Biology

Localized RNAi and Ectopic Gene Expression in the Medicinal Leech

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.

Biology

Spinal Cord Electrophysiology

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.

Education

Thermodynamics of Membrane Protein Folding Measured by Fluorescence Spectroscopy

Diana E. Schlamadinger¹, Judy E. Kim¹

¹Chemistry 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.

Medicine

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

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.

Neuroscience

Using Affordable LED Arrays for Photo-Stimulation of Neurons

Matthew Valley¹, Sebastian Wagner¹, Benjamin W. Gallarda¹, Pierre-Marie Lledo¹

¹Laboratory for Perception and Memory, Institut Pasteur and Centre National de la Recherche Scientifique (CNRS)

Adult-born neurons expressing ChR2 can be manipulated in slice electrophysiological preparations in order to examine their contribution towards the function of olfactory neural circuits.

Neuroscience

Selective Viral Transduction of Adult-born Olfactory Neurons for Chronic in vivo Optogenetic Stimulation

Gabriel Lepousez¹, Mariana Alonso¹, Sebastian Wagner¹, Benjamin W. Gallarda¹, Pierre-Marie Lledo¹

¹Laboratory for Perception and Memory, Institut Pasteur and Centre National de la Recherche Scientifique (CNRS)

Adult-born neurons of the olfactory bulb can be optogenetically controlled using Channelrhodopsin2-expressing lentiviral injection in the rostral migratory stream and chronic photostimulation with an implanted miniature LED.

Bioengineering

Designing Porous Silicon Films as Carriers of Nerve Growth Factor

Michal Rosenberg¹, Neta Zilony^2,3, Orit Shefi^2,3, Ester Segal^1,4

¹Department of Biotechnology and Food Engineering, Technion - Israel Institute of Technology, ²Faculty of Engineering, Bar-Ilan University, ³Bar-Ilan Institute of Nanotechnologies and Advanced Materials, ⁴Russell Berrie Nanotechnology Institute, Technion - Israel Institute of Technology

Here, we present a protocol to design and fabricate nanostructured porous silicon (PSi) films as degradable carriers for the nerve growth factor (NGF). Neuronal differentiation and outgrowth of PC12 cells and mice dorsal root ganglion (DRG) neurons are characterized upon treatment with the NGF-loaded PSi carriers.

Bioengineering

Fabrication of Magnetic Platforms for Micron-Scale Organization of Interconnected Neurons

Ganit Indech^1,4, Reut Plen^2,4, Dafna Levenberg^2,4, Naor Vardi^1,4, Michal Marcus^2,4, Alejandra Smith^2,4, Shlomo Margel^3,4, Orit Shefi^2,4, Amos Sharoni^1,4

¹Department of Physics, Bar-Ilan University, ²Faculty of Engineering, Bar-Ilan University, ³Department of chemistry, Bar-Ilan University, ⁴The Institutes of Nanotechnology & Advanced Materials, Bar-Ilan University

This work presents a bottom-up approach to the engineering of local magnetic forces for control of neuronal organization. Neuron-like cells loaded with magnetic nanoparticles (MNPs) are plated atop and controlled by a micro-patterned platform with perpendicular magnetization. Also described are magnetic characterization, MNP cellular uptake, cell viability, and statistical analysis.