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The Salk Institute for Biological Studies

9 ARTICLES PUBLISHED IN JoVE

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Biology

Spinal Cord Electrophysiology
Allyn Meyer 1, Benjamin W. Gallarda 1,2, Samuel Pfaff 1, William Alaynick 1
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.

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Biology

Mutagenesis and Functional Analysis of Ion Channels Heterologously Expressed in Mammalian Cells
Bartosz Balana 1, Natalie Taylor 1, Paul A. Slesinger 1
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.

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Neuroscience

Organotypic Cerebellar Cultures: Apoptotic Challenges and Detection
Tatiana Hurtado de Mendoza 1, Bartosz Balana 2, Paul A. Slesinger 2, Inder M. Verma 1
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.

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Neuroscience

Spinal Cord Electrophysiology II: Extracellular Suction Electrode Fabrication
Suresh Garudadri 1, Benjamin Gallarda 1, Samuel Pfaff 1, William Alaynick 1
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

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Neuroscience

Isolating Nasal Olfactory Stem Cells from Rodents or Humans
Stéphane D. Girard 1,2, Arnaud Devéze 3, Emmanuel Nivet 1,2,4, Bruno Gepner 5, François S. Roman 2, 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.

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Neuroscience

Construction of Cell-based Neurotransmitter Fluorescent Engineered Reporters (CNiFERs) for Optical Detection of Neurotransmitters In Vivo
Emre Lacin 1, Arnaud Muller 2, Marian Fernando 1, David Kleinfeld 2,3, Paul A. Slesinger 1
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.

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Neuroscience

Optical Control of a Neuronal Protein Using a Genetically Encoded Unnatural Amino Acid in Neurons
Ji-Yong Kang 1, Daichi Kawaguchi 2, Lei Wang 3
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.

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Education

External Excitation of Neurons Using Electric and Magnetic Fields in One- and Two-dimensional Cultures
Shani Stern 1, Assaf Rotem 2, Yuri Burnishev 3, Eyal Weinreb 3, Elisha Moses 3
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.

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Biochemistry

Single-Particle Cryo-EM Data Collection with Stage Tilt using Leginon
Sriram Aiyer 1, Timothy S. Strutzenberg 1, Marianne E. Bowman 2, 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.

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