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.
1Biological Sciences, University of Illinois at Chicago
Recording Ca2+ currents at the presynaptic release face membrane is key to a precise understanding of Ca2+ entry and neurotransmitter release. We present an acute dissociation of the lamprey spinal cord that yields functional isolated reticulospinal axons, permitting recording directly from the release face membrane of individual presynaptic terminals.
Published October 1, 2014. Keywords: Neuroscience, reticulospinal synapse, reticulospinal axons, presynaptic terminal, presynaptic calcium, voltage-gated calcium channels, vesicle fusion, synaptic transmission, neurotransmitter release, spinal cord, lamprey, synaptic vesicles, acute dissociation
1Institute of Integrative Neuroanatomy, NeuroCure Cluster of Excellence, Charité Universitätmedizin
Cortical networks are controlled by a small, but diverse set of inhibitory interneurons. Functional investigation of interneurons therefore requires targeted recording and rigorous identification. Described here is a combined approach involving whole-cell recordings from single or synaptically-coupled pairs of neurons with intracellular labeling, post-hoc morphological and immunocytochemical analysis.
Published September 30, 2014. Keywords: Neuroscience, electrophysiology, acute slice, whole-cell patch-clamp recording, neuronal morphology, immunocytochemistry, parvalbumin, hippocampus, inhibition, GABAergic interneurons, synaptic transmission, IPSC, GABA-B receptor
1Department of Physiology and Centre for Brain Research, University of Auckland, 2Department of Molecular and Cellular Physiology, Stanford University
Pair recordings are simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling precise electrophysiological and pharmacological characterization of the synapses between individual neurons. Here we describe the detailed methodology and requirements for establishing this technique in organotypic hippocampal slice cultures in any laboratory equipped for electrophysiology.
Published September 28, 2014. Keywords: Neuroscience, hippocampus, paired recording, whole cell recording, organotypic slice, synapse, synaptic transmission, synaptic plasticity
1Department of Natural Sciences, Assumption College
Cell culture models provide detailed control over environmental conditions and thus provide a powerful platform to elucidate numerous aspects of neuronal cell biology. We describe a rapid, inexpensive, and reliable method to isolate, dissociate, and culture sensory neurons from chick embryos. Details of substrata preparation and immunocytochemistry are also provided.
Published September 24, 2014. Keywords: Neuroscience, dorsal root gangia, DRG, chicken, in vitro, avian, laminin-1, embryonic, primary
1Department of Mechanical Engineering, San José State University, 2J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, 3Department of Physiology, Emory University School of Medicine, 4Department of Biological Sciences, San José State University
Muscle sensory neurons are involved in proprioceptor signaling and also report on metabolic state and injury related events. We describe an adult mouse in vitro muscle-nerve preparation for studies on stretch-activated muscle afferents.
Published September 24, 2014. Keywords: Neuroscience, muscle spindle, muscle afferent, extensor digitorum longus, sensory neurons, electrophysiology
1Cancer Biology Program, Fox Chase Cancer Center, Temple University Health System
Nestin-expressing progenitors are a newly identified population of neuronal progenitors in the developing cerebellum. Using the microdissection technique presented here in combination with fluorescent-activated cell sorting, this cell population can be purified with no contamination from other cerebellar regions and can be cultured for further studies.
Published September 21, 2014. Keywords: Neuroscience, microdissection, cerebellum, EGL, Nestin, medulloblastoma
1Centre de Résonance Magnétique Biologique et Médicale, UMR 7339 CNRS, Faculté de Médecine, Aix-Marseille Université, 2Centre de Recherches en Oncologie Biologique et Oncopharmacologie, UMR 911 INSERM, Faculté de Médecine, Aix-Marseille Université
The neurochemistry of mammalian brain is changed in many neurological and systemic diseases. Characteristic profiles of cerebral metabolites can be efficiently obtained based on crude extracts of brain tissue. To this end, high-resolution NMR spectroscopy is employed, enabling detailed quantitative analysis of metabolite concentrations (metabolomics).
Published September 21, 2014. Keywords: Neuroscience, metabolomics, brain tissue, rodents, neurochemistry, tissue extracts, NMR spectroscopy, quantitative metabolite analysis, cerebral metabolism, metabolic profile
1Department of Neurosciences, University of California, San Diego, 2School of Biomedical Engineering and Med-X Research Institute, Shanghai Jiao Tong University, 3Department of Anesthesiology, University of California, San Diego, 4VA San Diego Healthcare System
Axonal transport of BDNF, a neurotrophic factor, is critical for the survival and function of several neuronal populations. Some degenerative disorders are marked by disruption of axonal structure and function. We demonstrated the techniques used to examine live trafficking of QD-BDNF in microfluidic chambers using primary neurons.
Published September 15, 2014. Keywords: Neuroscience, live imaging, brain-derived neurotrophic factor (BDNF), quantum dot, trafficking, axonal retrograde transport, microfluidic chamber
1Department of Neuroscience, Baylor College of Medicine, 2Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease, University of Florida, 3Department of Neurology and Department of Neurosurgery, Baylor College of Medicine
Here we demonstrate a technique for widespread neuronal transduction by intraventricular injection of adeno-associated virus into the neonatal mouse brain. This method provides a rapid and easy way to attain lifelong expression of virally-delivered transgenes.
Published September 15, 2014. Keywords: Neuroscience, AAV, adeno-associated virus, viral transduction, neuronal transduction, intraventricular injection, neonatal injection, brain transgenesis, viral labeling
1Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, 2Department of Obstetrics and Gynecology, Stanford University School of Medicine
We, based on knowledge from developmental biology and published research, developed an optimized protocol to efficiently generate A9 midbrain dopaminergic neurons from both human embryonic stem cells and human induced pluripotent stem cells, which would be useful for disease modeling and cell replacement therapy for Parkinson’s disease.
Published September 15, 2014. Keywords: Neuroscience, dopaminergic neuron, substantia nigra pars compacta, midbrain, Parkinson’s disease, directed differentiation, human pluripotent stem cells, floor plate