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The Salk Institute for Biological Studies

9 ARTICLES PUBLISHED IN JoVE

Biology

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.

Biology

Mutagenesis and Functional Analysis of Ion Channels Heterologously Expressed in Mammalian Cells

Bartosz Balana¹, Natalie Taylor¹, Paul A. Slesinger¹

¹Clayton Foundation Laboratories for Peptide Biology, Salk Institute for Biological Studies

We will demonstrate how to study the effect of a single point mutation on the function of an ion channel.

Neuroscience

Organotypic Cerebellar Cultures: Apoptotic Challenges and Detection

Tatiana Hurtado de Mendoza¹, Bartosz Balana², Paul A. Slesinger², Inder M. Verma¹

¹Laboratory of Genetics, The Salk Institute for Biological Studies, ²Clayton Foundation Laboratories for Peptide Biology, The Salk Institute for Biological Studies

This method describes the generation of organotypic cerebellar cultures and the effect of certain apoptotic stimuli on the viability of different cerebellar cell types.

Neuroscience

Spinal Cord Electrophysiology II: Extracellular Suction Electrode Fabrication

Suresh Garudadri¹, Benjamin Gallarda¹, Samuel Pfaff¹, William Alaynick¹

¹Howard Hughes Medical Institute and Gene Expression Laboratory, The Salk Institute for Biological Studies

A demonstration of the fabrication and use of an extracellular suction electrode used to measure electrophysiological recordings of neonatal rodent spinal cords in vitro

Neuroscience

Isolating Nasal Olfactory Stem Cells from Rodents or Humans

Stéphane D. Girard^1,2, Arnaud Devéze³, Emmanuel Nivet^1,2,4, Bruno Gepner⁵, François S. Roman², François Féron^1,6

¹NICN, Aix Marseille University, ²LNPM, Aix Marseille University, ³ENT Department, Aix Marseille University, ⁴Gene expression Laboratory, The Salk Institute for Biological Studies, ⁵Laboratory of Speech and Language, Aix Marseille University, ⁶Centre d'Investigations Cliniques en Biothérapie, Aix Marseille University

We describe here a method for biopsying olfactory mucosa from rat and human nasal cavities. These biopsies can be used for either identifying molecular anomalies in brain diseases or isolating multipotent adult stem cells that can be utilized for cell transplantation in animal models of brain trauma/disease.

Neuroscience

Construction of Cell-based Neurotransmitter Fluorescent Engineered Reporters (CNiFERs) for Optical Detection of Neurotransmitters In Vivo

Emre Lacin¹, Arnaud Muller², Marian Fernando¹, David Kleinfeld^2,3, Paul A. Slesinger¹

¹Department of Neuroscience, Icahn School of Medicine at Mount Sinai, ²Department of Physics, University of California, San Diego, ³Section of Neurobiology, University of California, San Diego

We present a protocol to create cell-based neurotransmitter fluorescent engineered reporters (CNiFERs) for the optical detection of volumetric neurotransmitter release.

Neuroscience

Optical Control of a Neuronal Protein Using a Genetically Encoded Unnatural Amino Acid in Neurons

Ji-Yong Kang¹, Daichi Kawaguchi², Lei Wang³

¹Department of Neuroscience, School of Medicine, Tufts University, ²Molecular Neurobiology Laboratory, The Salk Institute for Biological Studies, ³Department of Pharmaceutical Chemistry and the Cardiovascular Research Institute, University of California, San Francisco

Here, a procedure to selectively activate a neuronal protein with a short pulse of light by genetically encoding a photo-reactive unnatural amino acid into a target neuronal protein expressed in neurons in culture or in vivo is presented.

Education

External Excitation of Neurons Using Electric and Magnetic Fields in One- and Two-dimensional Cultures

Shani Stern¹, Assaf Rotem², Yuri Burnishev³, Eyal Weinreb³, Elisha Moses³

¹Laboratory of Genetics, The Salk Institute for Biological Studies, ²Department of Physics and SEAS, Harvard University, ³Department of Physics of Complex Systems, Weizmann Institute of Science

Neuronal cultures are a good model for studying emerging brain stimulation techniques via their effect on single neurons or a population of neurons. Presented here are different methods for stimulation of patterned neuronal cultures by an electric field produced directly by bath electrodes or induced by a time-varying magnetic field.

Biochemistry

Single-Particle Cryo-EM Data Collection with Stage Tilt using Leginon

Sriram Aiyer¹, Timothy S. Strutzenberg¹, Marianne E. Bowman², Joseph P. Noel^2,3, Dmitry Lyumkis^1,4,5

¹Laboratory of Genetics, The Salk Institute for Biological Studies, ²Jack H. Skirball Center for Chemical Biology and Proteomics, The Salk Institute for Biological Studies, ³Department of Chemistry and Biochemistry, University of California San Diego, ⁴Graduate School of Biological Sciences, Section of Molecular Biology, University of California San Diego, ⁵Department of Integrative Structural and Computational Biology, The Scripps Research Institute

The present protocol describes a generalized and easy-to-implement scheme for tilted single-particle data collection in cryo-EM experiments. Such a procedure is especially useful for obtaining a high-quality EM map for samples suffering from preferential orientation bias due to adherence to the air-water interface.