JoVE Neuroscience is a multidisciplinary section devoted to investigations of the structure, function, physiology, and pathophysiology of the brain and nervous system. Included methodologies range from molecular and cellular level studies to full central and peripheral neural systems. Potential treatment platforms and surgical techniques for neurological diseases and disorders are also presented in this section.
1Department of Cell Biology and Physiology, Washington University School of Medicine
Four-dimensional (4D) imaging is utilized to study the behavior and interactions among two types of endosomes in living vertebrate nerve terminals. Movement of these small structures is characterized in three dimensions, permitting confirmation of events such as endosome fusion and exocytosis.
Published April 16, 2014. Keywords: Neuroscience, Microscopy, Fluorescence, Endocytosis, nerve, endosome, lysosome, deconvolution, 3D, 4D, epifluorescence
1Department of Veterans Affairs, Minneapolis Veterans Affairs Health Care System, 2Department of Food Science and Nutrition, University of Minnesota, 3Department of Integrative Biology and Physiology, University of Minnesota, 4Department of Medicine, University of Minnesota Medical School, University of Minnesota
Multiplex assays can provide beneficial information for basic cellular mechanisms and eliminate waste of reagents and unnecessary repetitive experiments. We describe here a multiplex caspase-3/7 activity assay, using fluorescent- and luminescent-based methods, to determine cell viability in an in vitro hypothalamic model following oxidative challenge with palmitic acid.
Published April 16, 2014. Keywords: Neuroscience, apoptosis, obesity, caspase, resazurin, DEVD, palmitic acid, hypothalamic model
1Leibniz Institute for Age Research, Fritz Lipmann Institute, 2Friedrich Schiller University Jena, 3Institute of Molecular Cell Biology & Center for Sepsis Control and Care (CSCC) Jena University Hospital, Friedrich Schiller University Jena
Measurements of nerve conduction properties in vivo exemplify a powerful tool to characterize various animal models of neuromuscular diseases. Here, we present an easy and reliable protocol by which electrophysiological analysis on sciatic nerves of anesthetized mice can be performed.
Published April 13, 2014. Keywords: Neuroscience, Demyelinating Diseases, Neurodegenerative Diseases, electrophysiology, sciatic nerve, mouse, nerve conduction velocity, neuromuscular diseases
1Genetics and Developmental Biology, School of Medicine, University of Connecticut Health Center
Drosophila melanogaster are useful in studying genetic or environmental manipulations that affect behaviors such as spontaneous locomotor activity. Here we describe a protocol that utilizes monitors with infrared beams and data analysis software to quantify spontaneous locomotor activity.
Published April 10, 2014. Keywords: Neuroscience, Investigative Techniques, Life Sciences (General), Behavioral Sciences, Drosophila melanogaster, Fruit flies, Spontaneous physical activity, Mobility, Fly behavior, Locomotor Activity
1Team Alzheimer & Tauopathies, Jean-Pierre Aubert Research Centre, Inserm UMR 837, 2EA 4308-Department of Reproductive Biology-Spermiology-CECOS, CHRU-Lille, 3EA2686-Laboratorie d'Immunologie, Faculté de Médecine - Pôle Recherche, 4Department of Neurology, CHRU-Lille
A common protein extraction protocol using urea/thiourea/SDS buffer for human and mice brain tissue allows indentification of proteins by 2D-DIGE and their subsequent characterization by mini 2DE immunoblotting. This method enables one to obtain more reproducible and reliable results from human biopsies and experimental models.
Published April 10, 2014. Keywords: Neuroscience, proteomics, neurodegeneration, 2DE, human and mice brain tissue, fluorescence, immunoblotting.
Abbreviations: 2DE (two-dimensional gel electrophoresis), 2D-DIGE (two-dimensional fluorescence difference gel electrophoresis), mini-2DE (mini 2DE immunoblotting), IPG (Immobilized pH Gradients), IEF (isoelectrofocusing), AD (Alzheimer´s disease)
1Program in Neuroscience, Graduate School of Biomedical Sciences, University of Massachusetts Medical School, 2Brudnick Neuropsychiatric Research Institute, Department of Psychiatry, University of Massachusetts Medical School
Neuronal membrane trafficking dynamically controls plasma membrane protein availability and significantly impacts neurotransmission. To date, it has been challenging to measure neuronal endocytic trafficking in adult neurons. Here, we describe a highly effective, quantitative method to measure rapid changes in surface protein expression ex vivo in acute brain slices.
Published April 3, 2014. Keywords: Neuroscience, Trafficking, endocytosis, internalization, biotinylation, brain, neurons, transporter, protein kinase C
1Department of Biology, Case Western Reserve University
We previously developed a technique for implanting tetrode wires into the central complex of cockroach brains that allows us to monitor activity in individual units of tethered cockroaches. Here we present a modified version of that technique that allows us to also record brain activity in freely moving insects.
Published April 1, 2014. Keywords: Neuroscience, Central complex, Free walking, Climbing, Brain recording, Tetrode, Fan-shaped body
1Department of Molecular Physiology & Biophysics, University of Iowa Carver College of Medicine, 2Department of Biology & Biochemistry, University of Bath
We describe the use of styryl FM dyes to image synaptic vesicle recycling in functional nerve terminals. This protocol can be applied not only to evoked, but also spontaneous and miniature synaptic activities. The protocol expands the variety of synaptic events that can be effectively evaluated.
Published March 31, 2014. Keywords: Neuroscience, Presynaptic Terminals, Synaptic Vesicles, Microscopy, Biological Assay, Nervous System, Endocytosis, exocytosis, fluorescence imaging, FM dye, neuron, photobleaching
1Hans Berger Department of Neurology, Jena University Hospital, Jena, Germany, 2Integrated Research and Treatment Center, Center for Sepsis Control and Care (CSCC), Jena University Hospital, Jena, Germany
GABAergic presynaptic inhibition is a powerful inhibitory mechanism in the spinal cord important for motor and sensory signal integration in spinal cord networks. Underlying primary afferent depolarization can be measured by recording of dorsal root potentials (DRP). Here we demonstrate a method of in vivo recording of DRP in mice.
Published March 29, 2014. Keywords: Neuroscience, Central Nervous System Diseases, Spinal Cord Diseases, Electrophysiology, dorsal root potentials (DRP), spinal cord, GABA, presynaptic inhibition, primary afferent depolarization (PAD), in vivo electrophysiology
1Pharmacology & Chemical Biology Department, University of Pittsburgh School of Medicine
Here we demonstrate the use of fluorescent Alexa dye coupled to α-bungarotoxin to measure GABA A receptor surface localization and endocytosis in hippocampal neurons. Through the use of constructs bearing a short extracellular tag that binds α-bungarotoxin, analysis of plasma membrane protein endocytic trafficking can be achieved.
Published March 28, 2014. Keywords: Neuroscience, α-bungarotoxin, binding site, endocytosis, immunostaining, rodent hippocampal neurons, receptor, trafficking, plasma membrane