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UCL Institute of Neurology

9 ARTICLES PUBLISHED IN JoVE

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Biology

Assaying the Kinase Activity of LRRK2 in vitro
Patrick A. Lewis 1
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.

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Medicine

Controlling Parkinson's Disease With Adaptive Deep Brain Stimulation
Simon Little 1, Alek Pogosyan 1, Spencer Neal 2, Ludvic Zrinzo 2, Marwan Hariz 2, Thomas Foltynie 2, Patricia Limousin 2, Peter Brown 1
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.

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Neuroscience

Sequential Extraction of Soluble and Insoluble Alpha-Synuclein from Parkinsonian Brains
Rina Bandopadhyay 1
1Reta Lila Weston Institute of Neurological Studies, UCL Institute of Neurology

Here, we present a protocol for the isolation of increasingly insoluble/aggregated alpha-synuclein (α-syn) from post-mortem human brain tissue. Through the utilization of buffers with increasing detergent strength and high-speed ultracentrifugation techniques, the variable properties of α-syn aggregation in diseased and non-diseased tissue can be examined.

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Medicine

A Pipeline for 3D Multimodality Image Integration and Computer-assisted Planning in Epilepsy Surgery
Mark Nowell 1, Roman Rodionov 1, Gergely Zombori 2, Rachel Sparks 2, Michele Rizzi 1, Sebastien Ourselin 2, Anna Miserocchi 3, Andrew McEvoy 3, John Duncan 1
1Department of Clinical and Experimental Epilepsy, UCL Institute of Neurology, 2Center of Medical Imaging and Computing, UCL, 3Department of Neurosurgery, National Hospital for Neurology and Neurosurgery

We describe the steps to use our custom designed software for image integration, visualization and planning in epilepsy surgery.

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Research

Measuring Lactase Enzymatic Activity in the Teaching Lab
Cattleya S. Leksmono 1, Claudia Manzoni 1,2, James E. Tomkins 1, Walter Lucchesi 1,3, Graeme Cottrell 1, Patrick A. Lewis 1
1School of Pharmacy, University of Reading, 2Department of Molecular Neuroscience, UCL Institute of Neurology, 3School of Biological Sciences, Royal Hollway, University of London

The enzymatic activity of lactase is essential for the catabolic processing of the disaccharide lactose. Here, the activity of lactase found in dietary supplements is assayed using a colorimetric assay. This provides students with an experimental platform for understanding the activity of lactase and enzyme kinetics.

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Medicine

A High Throughput, Multiplexed and Targeted Proteomic CSF Assay to Quantify Neurodegenerative Biomarkers and Apolipoprotein E Isoforms Status
Wendy E. Heywood *1, Anna Baud *1, Emily Bliss 1, Ernestas Sirka 1, Jonathan M. Schott 2, Henrik Zetterberg 3, Daniela Galimberti 4, Neil J. Sebire 5, Kevin Mills 1
1Centre for Translational Omics, Genetics and Genomic Medicine Deptartment, Great Ormond Street Institute of Child Health, University College London, 2Dementia Research Centre, Institute of Neurology, University College London, 3Clinical Neurochemistry Laboratory, Institute of Neuroscience and Physiology, Department of Psychiatry and Neurochemistry, The Sahlgrenska Academy, University of Gothenburg, 4Neurology Unit, Department of Pathophysiology and Transplantation, University of Milan, 5Great Ormond Street Hospital for Children, University College London

We describe a high-throughput, multiplex, and targeted proteomic cerebrospinal fluid (CSF) assay developed with potential for clinical translation. The test can quantitate potential markers and risk factors for neurodegeneration, such as the apolipoprotein E variants (E2, E3 and E4), and measure their allelic expression.

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Medicine

Absolute Quantification of Aβ1-42 in CSF Using a Mass Spectrometric Reference Measurement Procedure
Josef Pannee 1,2, Kaj Blennow 1,2, Henrik Zetterberg 1,2,3, Erik Portelius 1,2
1Institute of Neuroscience and Physiology, Department of Psychiatry and Neurochemistry, The Sahlgrenska Academy at University of Gothenburg, Sahlgrenska University Hospital, 2Clinical Neurochemistry Laboratory, Sahlgrenska University Hospital, 3UCL Institute of Neurology, Queen Square

A reference measurement procedure for the absolute quantification of Aβ1-42 in human CSF based on solid-phase extraction and liquid chromatography tandem mass spectrometry is described.

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Neuroscience

Sample Preparation for Endopeptidomic Analysis in Human Cerebrospinal Fluid
Karl T. Hansson 1, Tobias Skillbäck 1,2, Elin Pernevik 1, Jessica Holmén-Larsson 1, Gunnar Brinkmalm 1, Kaj Blennow 1,2, Henrik Zetterberg 1,2,3, Johan Gobom 1,2
1Inst. of Neuroscience and Physiology, Dept. of Psychiatry and Neurochemistry, Sahlgrenska Academy at University of Gothenburg, 2Clinical Neurochemistry Laboratory, Sahlgrenska University Hospital, 3Department of Molecular Neuroscience, UCL Institute of Neurology

A method for mass spectrometric analysis of endogenous peptides in human cerebrospinal fluid (CSF) is presented. By employing molecular weight cut-off filtration, chromatographic pre-fractionation, mass spectrometric analysis and a subsequent combination of peptide identification strategies, it was possible to expand the known CSF peptidome nearly ten-fold compared to previous studies.

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Neuroscience

Automated Segmentation of Cortical Grey Matter from T1-Weighted MRI Images
Eileanoir B. Johnson 1, Rachael I. Scahill 1, Sarah J. Tabrizi 1
1Huntington's Disease Research Centre, UCL Institute of Neurology

This protocol describes the process of applying seven different automated segmentation tools to structural T1-weighted MRI scans to delineate grey matter regions that can be used for the quantification of grey matter volume.

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