1Center for Visual and Neurocognitive Rehabilitation, Atlanta VAMC, Department of Medicine, Division of General Medicine and Geriatrics, Emory University School of Medicine, 2Harvard Neurology Residency Program, Brigham and Woman‘s Hospital and Massachusetts General Hospital
Adapted tango has demonstrated efficacy for improving mobility and balance. We describe the dissemination of adapted tango teaching methods to dance instructor trainees and the implementation of adapted tango by the trainees in the community for improving mobility and balance in older adults and individuals with Parkinson’s disease.
Published December 9, 2014. Keywords: Behavior, Dance, tango, balance, pedagogy, dissemination, exercise, older adults, Parkinson's Disease, mobility impairments, falls
1Center for Health Sciences, SRI International, 2Department of Chemistry and Biochemistry, University of California-Santa Cruz
Lipoxygenase (LOX) isozymes can generate products that may increase or decrease neuroinflammation and neurodegeneration. A gene-environment interaction study could identify LOX isozyme-specific effects. Using the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) model of nigrostriatal damage in two LOX isozyme-deficient transgenic lines allows for comparison of the contribution of LOX isozymes on dopaminergic integrity and inflammation.
Published January 7, 2014. Keywords: Medicine, MPTP, dopamine, Iba1, TH, GFAP, lipoxygenase, transgenic, gene-environment interactions, mouse, Parkinson's disease, neurodegeneration, neuroinflammation
1Centre for Neurobiology of Stress, Dept Biological Sciences, University of Toronto at Scarborough
A protocol for performing unilateral 6-OHDA lesions of the medial forebrain bundle in mice is described. This method has a low mortality rate (13.3 %) with 89% of the surviving animals showing >95% loss of striatal dopamine and 90.63±-4.02 % ipsiversive rotational bias towards the side of the lesion.
Published February 14, 2012. Keywords: Medicine, mouse, 6-OHDA, Parkinson’s disease, medial forebrain bundle, unilateral
1Department of Biology, Colby College
Parkinson’s disease is a neurodegenerative disorder that results from the degeneration of dopaminergic neurons in the central nervous system, causing locomotion defects. Rotenone models Parkinson’s disease in Drosophila. This paper outlines two assays that characterize both spontaneous and startle-induced locomotion deficiencies caused by rotenone.
Published August 17, 2014. Keywords: Neuroscience, Locomotion, Parkinson’s disease, rotenone, Drosophila, activity monitoring, neurobiology, behavior
1Nuffield Department of Clinical Neurosciences, John Radcliffe Hospital, University of Oxford, 2Sobell Department of Motor Neuroscience & Movement Disorders, Unit of Functional Neurosurgery, UCL Institute of Neurology
Adaptive deep brain stimulation (aDBS) is effective for Parkinson’s disease, improving symptoms and reducing power consumption compared to conventional deep brain stimulation (cDBS). In aDBS we track a local field potential biomarker (beta oscillatory amplitude) in real time and use this to control the timing of stimulation.
Published July 16, 2014. Keywords: Medicine, Parkinson's, deep brain stimulation, adaptive, closed loop
1Department of Psychology, University of Cincinnati, 2Department of Neurology, University of Cincinnati
In Parkinson's disease and movement disorders in general, sensitive and reliable behavioral assays are essential for testing novel potential therapeutics. Here, we describe a manageable battery of sensorimotor tests for mice that are sensitive to varying degrees of injury to the nigrostriatal system and useful for preclinical studies.
Published June 17, 2013. Keywords: Behavior, Neuroscience, Neurobiology, Medicine, Biomedical Engineering, Anatomy, Physiology, Psychology, Basal Ganglia Diseases, Parkinsonian Disorders, Parkinson Disease, Genetics, Behavioral, Psychopharmacology, sensory, motor, mouse, movement disorders, beam, cylinder, animal model
1German Center for Neurodegenerative Diseases, DZNE, 2Laboratory of Functional Neurogenomics, Department of Neurodegenerative Diseases, Hertie Institute for Clinical Brain Research, University of Tübingen
Fibroblasts from patients carrying mutations in Parkinson's disease-causing genes represent an easily accessible ex vivo model to study disease-associated phenotypes. Live cell imaging gives the opportunity to study morphological and functional parameters in living cells. Here we describe the preparation of human fibroblasts and subsequent monitoring of mitochondrial phenotypes.
Published October 3, 2012. Keywords: Medicine, Genetics, Cellular Biology, Physiology, Parkinson's disease, fibroblasts, mitochondria, live cell imaging, mitochondrial function, mitochondrial morphology, mitophagy
1Department of Pharmaceutical Biosciences, Uppsala University, 2Department of Chemical and Biological Engineering, Chalmers University of Technology
Dopamine replacement pharmacotherapy using L-DOPA is the most commonly used symptomatic treatment of Parkinson’s disease, but is accompanied by side effects including involuntary abnormal movements, termed dyskinesia 1. Here, a protocol for MALDI imaging mass spectrometry is presented that detects changes in rat brain neuropeptide levels related to dyskinesia.
Published February 14, 2012. Keywords: Medicine, Parkinson's disease, L-DOPA induced dyskinesia, striatum, opioid peptides, MALDI Imaging MS
1Department of Biological Sciences, University of Alabama
This video demonstrates how to use C. elegans to assess dopaminergic neuron neurodegeneration as a model for Parkinson's disease. Furthermore, genetic screens are used to identify factors that either enhance degeneration or are neuroprotective.
Published July 18, 2008. Keywords: Neuroscience, C. elegans, Parkinson's disease, neuroprotection, alpha-synuclein, Translational Research
1Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Brasil
Parkinson disease is caused by loss of dopaminergic innervation to the striatum, which can be experimentally induced by 6-OH-dopamine. We describe how to perform a stereotaxic lesion and to monitor apomorphine-induced rotational behavior in mice. This model is useful and reliable for testing new therapies for Parkinson disease.
Published January 15, 2010. Keywords: Neuroscience, neurodegenerative disease, mice, cell therapy, model
1McLean Hospital, Harvard Stem Cell Institute, Harvard Medical School
Ole Isacson gives a concise overview of Parkinsons's disease, its causes, therapeutic strategies, and advances in Parkinson's research.
Published April 29, 2007. Keywords: Medicine, Parkinson' disease, Neuroscience, dopamine, neuron, L-DOPA, stem cell, transplantation
1Center for Neurosciences, The Feinstein Institute for Medical Research
Multivariate techniques including principal component analysis (PCA) have been used to identify signature patterns of regional change in functional brain images. We have developed an algorithm to identify reproducible network biomarkers for the diagnosis of neurodegenerative disorders, assessment of disease progression, and objective evaluation of treatment effects in patient populations.
Published June 26, 2013. Keywords: Medicine, Neurobiology, Neuroscience, Anatomy, Physiology, Molecular Biology, Basal Ganglia Diseases, Parkinsonian Disorders, Parkinson Disease, Movement Disorders, Neurodegenerative Diseases, PCA, SSM, PET, imaging biomarkers, functional brain imaging, multivariate spatial covariance analysis, global normalization, differential diagnosis, PD, brain, imaging, clinical techniques
1Department of Neurology, Massachusetts General Hospital
We present a rapid and inexpensive screening method for identifying transcriptional regulators using high-throughput robotic transfections and a homemade dual-glow luciferase assay. This protocol rapidly generates direct side-by-side functional data for thousands of genes and is easily modifiable to target any gene of interest.
Published June 1, 2014. Keywords: Cellular Biology, Luciferases, Gene Transfer Techniques, Transfection, High-Throughput Screening Assays, Transfections, Robotics
1Laboratory for Neurobiology and Gene Therapy, Department of Neurosciences, KU Leuven and Leuven Institute for Neuroscience and Disease (LIND)
Leucine rich repeat kinases 1 and 2 (LRRK1 and LRRK2) are multidomain proteins which encode both GTPase and kinase domains and which are phosphorylated in cells. Here, we present a protocol to label LRRK1 and LRRK2 in cells with 32P orthophosphate, thereby providing a means to measure their overall cellular phophorylation levels.
Published September 18, 2013. Keywords: Cellular Biology, biology (general), biochemistry, bioengineering (general), LRRK1, LRRK2, metabolic labeling, 32P orthophosphate, immunoprecipitation, autoradiography
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
1CNRS, UMR-5203, Institut de Génomique Fonctionnelle, Montpellier, 2Inserm, U661, Montpellier, 3Universités de Montpellier
Dopaminergic neurons play a vital regulatory role in the brain. Their loss is associated with Parkinson's disease. In this video, we show how to generate primary cultures of central dopaminergic neurons from embryonic mouse mesencephalon. Such cultures are useful to study the extreme vulnerability of these neurons to various stresses.
Published September 8, 2014. Keywords: Neurobiology, Mus musculus, mesencephalon, embryonic, tyrosine hydroxylase, dopamine transporter, Parkinson's disease in vitro model
1Department of Psychology, University of Cincinnati, 2Department of Neurology, University of Cincinnati, 3Department of Neuroscience, Cell Biology, and Physiology, Wright State University
Impairment in olfactory function is a common feature in many neurodegenerative disorders including Parkinson, Alzheimer, and Huntington diseases. In the present article, we describe a set of tests for assessing olfaction discrimination and detection in mice that can be used to measure olfactory abilities in mouse models of neurodegenerative diseases.
Published August 25, 2014. Keywords: Neuroscience, olfaction, mouse, Parkinson’s disease, detection, discrimination, sniffing
1Department of Pharmacology, Toxicology, & Neuroscience, Louisiana State University Health Sciences Center
Exercise may alleviate behavioral impairments associated with neurodegenerative diseases. Studies of the mechanisms underlying exercise-mediated effects have increased in disease models, but the neurobiological background accompanying these diseases, specifically aging, is likely not considered. Here, we illustrate measures to increase the likelihood of a successful exercise paradigm in aged rats.
Published August 22, 2014. Keywords: Behavior, Exercise, locomotor, Parkinson’s disease, aging, treadmill, bradykinesia, Parkinsonism
1Division of Brain Sciences, Department of Medicine, Imperial College London, 2Department of Neurosurgery, Brigham and Women's Hospital, Harvard Medical School, 3Department of Internal Medicine, Endocrinology, Yale University School of Medicine
The causes of degeneration of midbrain dopaminergic neurons during Parkinson’s disease are not fully understood. Cellular culture systems provide an essential tool for study of the neurophysiological properties of these neurons. Here we present an optimized protocol, which can be utilized for in vitro modeling of neurodegeneration.
Published February 19, 2015. Keywords: Neuroscience, Ventral midbrain, Parkinson's disease, Dopaminergic, Primary neuronal culture, Neuronal development, Neurodegeneration
JoVE Immunology and Infection
1Department of Biological Sciences, University of Alabama
Levels of the inflammatory cell-signaling molecule nitric oxide (NO) are commonly assayed using Griess reagent. In this protocol, we have created a modified Griess assay utilizing live Drosophila brain tissue in order to detect the secretion of NO in a simple, quantifiable and highly repeatable method.
Published December 4, 2013. Keywords: Immunology, biology (general), environmental effects (biological, animal and plant), immunology, animal models, Immune System Diseases, Pathological Conditions, Signs and Symptoms, Life Sciences (General), Neuroinflammation, inflammation, nitric oxide, nitric oxide synthase, Drosophila, neurodegeneration, brain, Griess assay, nitrite detection, innate immunity, Parkinson disease, tissue culture
1School for Engineering of Matter, Transport and Energy, Arizona State University, 2Department of Neurology, Georgetown University Medical Center, 3Department of Pathology, Georgetown University Medical Center
Using atomic force microscopy in combination with biopanning technology we created a negative and positive biopanning system to acquire antibodies against disease-specific protein variants present in any biological material, even at low concentrations. We were successful in obtaining antibodies to TDP-43 protein variants involved in Amyotrophic Lateral Sclerosis.
Published February 12, 2015. Keywords: Bioengineering, Amyotrophic Lateral Sclerosis, TDP-43, Biopanning, Atomic Force Microscopy, scFv, Neurodegenerative diseases
1Department of Molecular Neuroscience, UCL Institute of Neurology
Leucine Rich Repeat Kinase 2 is a large multidomain kinase, mutations in which are the most common genetic cause of Parkinson's disease. Analysis of the kinase activity of this protein has proven to be a crucial tool in understanding the biology and dysfunction of this protein. In this paper, in vitro assaying of the kinase activity of LRRK2 and a selection of its mutants is described, providing an experimental system to examine phosphorylation of putative substrates and potential dysfunction of LRRK2 in disease.
Published January 18, 2012. Keywords: Molecular Biology, Kinase, LRRK2, Parkinson's disease
1Department of Surgery-Division of Otolaryngology, University of Wisconsin, 2Department of Communicative Disorders, University of Wisconsin
Voice disorders are debilitating in aging and Parkinson disease. The ultrasonic vocalizations of rats, also affected by these conditions, can be used to study these voice disorders, their neural substrates, and the nature of functional recovery with behavioral intervention.
Published August 8, 2011. Keywords: Neuroscience, ultrasonic vocalization, rat, aging, Parkinson disease, exercise, 6-hydroxydopamine, voice disorders, voice therapy
1Division of Pharmaceutical Sciences, Mylan School of Pharmacy, Duquesne University
Therapeutic compounds are often first examined in vitro with viability assays. Blind cell counts by a human observer can be highly sensitive to small changes in cell number but do not assess function. Computerized viability assays, as described here, can assess both structure and function in an objective manner.
Published January 20, 2014. Keywords: Cellular Biology, In-cell Western, DRAQ5, Sapphire, Cell Titer Glo, ATP, primary cortical neurons, toxicity, protection, N-acetyl cysteine, hormesis
1Department of Molecular Biosciences, Rice Institute for Biomedical Research, Northwestern University
Prion-like propagation of protein aggregates has recently emerged as being implicated in many neurodegenerative diseases. The goal of this protocol is to describe, how to use the nematode C. elegans as a model system to monitor protein spreading and to investigate prion-like phenomena.
Published January 8, 2015. Keywords: Cellular Biology, Caenorhabditis elegans, neurodegenerative diseases, protein misfolding diseases, prion-like spreading, cell-to-cell transmission, protein aggregation, non-cell autonomous toxicity, proteostasis
1School of Psychological Sciences, Tel-Aviv University, 2Sagol School of Neuroscience, Tel-Aviv University
The goal of the protocol described in this paper is to induce compulsive-like behavior in rats for the study of obsessive-compulsive disorder (OCD). This behavior is precipitated by attenuating a signal indicating that a lever-press response was effective in producing food.
Published January 9, 2015. Keywords: Behavior, Obsessive-compulsive disorder, OCD, signal attenuation, rat, animal model, pharmacology, lever-press, behavioral neuroscience
JoVE Immunology and Infection
1Department of Neuroscience, Georgetown University Medical Center
One key to successful investigation of microglial biology is the preservation of microglial immunofunction ex vivo during isolation from CNS tissue. Isolating microglia via rotary shaking results in highly pure and immunofunctional cell cultures as assessed by fluorescent imaging, immunocytochemistry, and ELISA following microglia activation with the proinflammatory stimuli lipopolysaccharide (LPS) and Pam3CSK4 (Pam).
Published January 30, 2014. Keywords: Immunology, neuroinflammation, Cytokines, neurodegeneration, LPS, Pam3CSK4, TLRs, PAMPs, DAMPs
1Nayef Al-Rodhan Laboratories for Cellular Neurosurgery and Neurosurgical Technology, Harvard Medical School, 2Department of Neurosurgery , Massachusetts General Hospital
Deep brain stimulation surgery offers a unique opportunity to examine information encoding in the awake human brain. This article will describe intra-operative methods used to perform cognitive and behavioral tasks while simultaneously acquiring physiological data such as EMG, single-unit neuronal activity and/or local field potentials.
Published January 6, 2011. Keywords: Medicine, Intra-Operative Physiology, Cognitive Neuroscience, Behavioral Neuroscience, Subthalamic Nucleus, Single-Unit Activity, Parkinson Disease, Deep Brain Stimulation
1Institute of Anatomy, Technische Universität Dresden
Parkinson's disease has been related to the exposure to pesticides. Here we show a method to deliver pesticides using a gastric tube at the desired concentration and a method to analyze their effect in alpha-synuclein accumulation in the enteric nervous system.
Published October 26, 2010. Keywords: Neuroscience, neurogical disorders, Parkinson's disease, animal model, mouse, rotenone, gavage, image analysis
1Ohio Musculoskeletal and Neurological Institute (OMNI) and the Department of Biomedical Sciences, Ohio University
Transcranial magnetic stimulation (TMS) is a non-invasive tool to gain insight on the physiology and function of the human nervous system. Here, we present our TMS techniques to study cortical excitability of the upper limb and lumbar musculature.
Published January 20, 2012. Keywords: Medicine, neuroscience, muscle, electromyography, physiology, TMS, strength, motor control. sarcopenia, dynapenia, lumbar
1Department of Biomolecular Genetics, University of Rochester Medical Center
Here we describe two assays that have been established to study age-dependent neurodegeneration of dopaminergic (DA) neurons in Drosophila: the climbing/startle-induced negative geotaxis assay which allows to study the functional effects of DA neurons degeneration and the tyrosine hydroxylase immunostaining which is used to identify and count DA neurons in whole brain mounts.
Published April 24, 2013. Keywords: Neuroscience, Genetics, Neurobiology, Molecular Biology, Cellular Biology, Biomedical Engineering, Medicine, Developmental Biology, Drosophila melanogaster, neurodegenerative diseases, negative geotaxis, tyrosine hydroxylase, dopaminergic neuron, α-synuclein, neurons, immunostaining, animal model
1Florey Institute of Neuroscience and Mental Health, The University of Melbourne
This protocol describes two different environmental manipulations and a concurrent brain infusion protocol to study environmentally-induced brain changes underlying adaptive behavior and brain repair in adult mice.
Published January 20, 2015. Keywords: Neuroscience, Behavior, midbrain, tyrosine hydroxylase, dopamine, plasticity, substantia nigra pars compacta
1Programa de Pós-Graduação em Ciências Médica, Universidade Federal do Rio Grande do Sul, 2Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES), 3Laboratory of Neuromodulation, Department of Physical Medicine & Rehabilitation, Spaulding Rehabilitation Hospital and Massachusetts General Hospital, Harvard Medical School, 4De Montfort University
Transcranial direct current stimulation (tDCS) is a non-invasive brain stimulation technique that has shown initial therapeutic effects in several neurological conditions. The main mechanism underlying these therapeutic effects is the modulation of cortical excitability. Therefore, online monitoring of cortical excitability would help guide stimulation parameters and optimize its therapeutic effects. In the present article we review the use of a novel device that combines simultaneous tDCS and EEG monitoring in real time.
Published June 17, 2013. Keywords: Behavior, Medicine, Neuroscience, Neurobiology, Anatomy, Physiology, Biomedical Engineering, Psychology, electroencephalography, electroencephalogram, EEG, transcranial direct current stimulation, tDCS, noninvasive brain stimulation, neuromodulation, closed-loop system, brain, imaging, clinical techniques
1Department of Biomedical Engineering, Boston University, 2Department of Otology and Laryngology, Massachusetts Eye and Ear Infirmary, Harvard Medical School
A mouse model of human endoscopic skull base reconstruction has been developed that creates a semipermeable interface between the brain and nose using nasal mucosal grafts. This method allows researchers to study delivery to the central nervous system of high molecular weight therapeutics which are otherwise excluded by the blood-brain barrier when administered systemically.
Published July 16, 2014. Keywords: Medicine, drug delivery, mucosa membrane, blood-brain barrier, neurosurgery, transnasal, mouse model
1University of Tennessee Health Science Center, 2Semmes-Murphey Clinic, 3University of Arkansas for Medical Sciences, 4Restorative Neurosciences Foundation
Convection-enhanced delivery (CED) has been proposed as a treatment option for a wide range of neurological diseases. In order to prepare health care professionals for adoption of CED, accessible training models are needed. We describe the use of agarose gel as such a model of the human brain for testing, research, and training.
Published May 14, 2014. Keywords: Medicine, Convection-enhanced delivery, agarose gel, volumes of distribution, gel infusion, Vd/Vi, MRI, Neurosurgery
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.
Published August 22, 2011. Keywords: Neuroscience, stem cell, nose, brain, neuron, cell therapy, diagnosis, sphere
1Department of Psychology and Neuroscience, Center for Neuroscience, University of Colorado Boulder, 2NeuroScript LLC, 3Department of Psychiatry, University of California, San Diego
Handwriting analysis software significantly improves upon existing instruments measuring movement disorders. Individuals at risk for psychosis and healthy controls completed handwriting tasks to test for dyskinesia. Results suggest that youth at risk for psychosis exhibit dyskinesia and that handwriting analysis could significantly contribute to wider dissemination of early identification efforts
Published November 21, 2013. Keywords: Behavior, Schizophrenia, Disorders with Psychotic Features, Psychology, Clinical, Psychopathology, behavioral sciences, Movement abnormalities, Ultra High Risk, psychosis, handwriting, computer tablet, dyskinesia
1Leibniz-Institut für Analytische Wissenschaften, ISAS, 2Department of Biochemical Engineering, University College London, 3Institute for Life Sciences, University of Southampton
Protocols for single neuron microfluidic arraying and water masking for the in-chip plasma patterning of biomaterial coatings are described. Highly interconnected co-cultures can be prepared using minimal cell inputs.
Published May 20, 2014. Keywords: Neuroscience, microfluidic arraying, single cell, biomaterial patterning, co-culture, compartmentalization, Alzheimer and Parkinson Diseases, neurite outgrowth, high throughput screening
1Emmy Noether-Group for Stem Cell Biology, Department of Molecular Embryology, Institute of Anatomy and Cell Biology, University of Freiburg, 2Spemann Graduate School of Biology and Medicine and Faculty of Biology, University of Freiburg, 3School of Life Sciences, Keele University, 4Center for Biological Signaling Studies (BIOSS), University of Freiburg
We provide a detailed description of a protocol for flow cytometric analysis of surface antigens and/or intracellular antigens in neural cell types. Critical aspects of experimental planning, step-by-step methodological procedures, and fundamental principles of flow cytometry are explained in order to enable neurobiologists to exploit this powerful technology.
Published December 18, 2014. Keywords: Neuroscience, CD markers, surface antigens, intracellular antigens, flow cytometry, neurons, glial cells, neural stem cells, fluorescence-activated cell sorting (FACS), CD24, CD54, CFSE
1Department of Biomedical Engineering, Oregon Health and Science University, 2UCL Institute of Neurology, Queen Square, 3Department of Neurology, Oregon Health and Science University
We have developed a device (Twister) to study the regulation of tonic muscle activity during active postural maintenance. Twister measures torsional resistance and muscular responses in standing subjects during twisting of the body axis. The device can be flexibly configured to study various aspects of tonic control across the neck, trunk, and/or hips.
Published December 14, 2011. Keywords: Medicine, Muscle Tone, Posture, Stiffness, Motor Control
1Department of Biomedical Engineering, The University of Texas at Austin, 2Department of Psychology, The University of Texas at Austin, 3The J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida
Three new behavioral tests (forelimb step-alternation, postural instability test, pasta handling test) for evaluating forelimb function after cervical spinal cord injury in rodents are described.
Published September 16, 2013. Keywords: Behavior, Behavior, Animal, Motor Activity, Nervous System Diseases, Wounds and Injuries, cervical spinal cord injury, lateral hemisection model, limb alternation, pasta handling, postural instability
1Department of Clinical Genetics, VU University Medical Center, 2Center for Neurogenomics and Cognitive Research, Neuroscience Campus Amsterdam
We describe a methodology combining automated cell culturing with high-content imaging to visualize and quantify multiple cellular processes and structures, in a high-throughput manner. Such methods can aid in the further functional annotation of genomes as well as identify disease gene networks and potential drug targets.
Published January 6, 2012. Keywords: Medicine, High-throughput screening, high-content screening, neurodegeneration, automated cell culturing, Parkinson’s disease
1Department of Biochemistry and Biophysics, Perelman School of Medicine at the University of Pennsylvania
Yeast proteinopathy models are valuable tools to assess the toxicity and aggregation of proteins implicated in disease. Here, we present methods for screening Hsp104 variant libraries for toxicity suppressors. This protocol could be adapted to screen any protein library for toxicity suppressors of any protein that is toxic in yeast.
Published November 11, 2014. Keywords: Microbiology, Protein-misfolding disorders, yeast proteinopathy models, Hsp104, proteotoxicity, amyloid, disaggregation
1Department of Neurology, University of Massachusetts Medical School
Standardized, comprehensive and fully quantitative testing of autonomic functions is described. The autonomic tests consist of evaluation of all three major autonomic domains including cardiovagal, adrenergic and sudomotor. The severity and distribution of dysautonomia is quantitated using Composite Autonomic Severity Scores.
Published July 19, 2011. Keywords: Medicine, Deep breathing, Valsalva maneuver, tilt test, sudomotor testing, Composite Autonomic Severity Score, CASS
1Department of Biological Sciences, University of Alabama
This video demonstrates the technique of microinjection into the gonad of C. elegans to create transgenic animals.
Published August 15, 2008. Keywords: Developmental Biology, C. elegans, microinjection, transgenic, stable lines
1Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, 2Quantitative Biology, Eli Lilly and Company
Persistent activation of inhibitory G protein-coupled receptors results in sensitization of adenylyl cyclase signaling. To identify the essential molecular pathways, nonbiased approaches are necessary; however, this strategy requires the development of a scalable cell-based cAMP sensitization assay. Herein, we describe a sensitization assay for small molecule and siRNA screening.
Published January 27, 2014. Keywords: Bioengineering, adenylyl cyclase, cAMP, heterologous sensitization, superactivation, D2 dopamine, μ opioid, siRNA
1School of Psychology, University of Birmingham
Transcranial direct current stimulation (tDCS) over the cerebellum exerts a remote effect on the prefrontal cortex, which can modulate cognition and performance. This was demonstrated using two information-processing tasks of varying complexity, whereby only cathodal tDCS improved performance when the task was difficult, but not easy.
Published February 15, 2015. Keywords: Behavior, Cognition, working memory, tDCS, cerebellum, brain stimulation, neuro-modulation, neuro-enhancement
1Department of Molecular Biotechnology and Health Science, University of Torino, 2Laboratory of Endothelial Molecular Biology, Vesalius Research Center, VIB
Here we report a protocol to measure oxidative stress in living zebrafish embryos. This procedure allows reactive oxygen species (ROS) detection in both whole embryo tissues and single-cell populations. This protocol will accomplish both qualitative and quantitative analyses.
Published July 7, 2014. Keywords: Developmental Biology, Danio rerio, zebrafish embryos, endothelial cells, redox state analysis, oxidative stress detection, in vivo ROS measurements, FACS (fluorescence activated cell sorter), molecular probes
1MRC/ARUK Centre for Musculoskeletal Ageing Research, University of Nottingham
Skeletal muscle is essential for locomotion and is the bodies’ main protein store. Muscle health measurements within C. elegans are described. Prospective changes to muscle structure and function are assessed using localized GFP and cationic dyes.
Published November 13, 2014. Keywords: Developmental Biology, Physiology, C. elegans, muscle, mitochondria, sarcomeres, ageing
1Department of Molecular Physiology and Biophysics, Baylor College of Medicine, 2Department of Neuroscience, Baylor College of Medicine, 3Department of Neuroscience, University of California at San Diego, 4National Center for Macromolecular Imaging, Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine
To preserve neuronal processes for ultrastructural analysis, we describe a protocol for plating of primary neurons on electron microscopy grids followed by flash freezing, yielding samples suspended in a layer of vitreous ice. These samples can be examined with a cryo-electron microscope to visualize structures at the nanometer scale.
Published February 12, 2014. Keywords: Neuroscience, Neurons, Cryo-electron Microscopy, Electron Microscope Tomography, Brain, rat, primary neuron culture, morphological assay