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University College Dublin

8 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Measurement & Analysis of the Temporal Discrimination Threshold Applied to Cervical Dystonia
Rebecca B Beck *1, Eavan M McGovern *1,2,3, John S Butler 4, Dorina Birsanu 1, Brendan Quinlivan 1, Ines Beiser 1,2,3, Shruti Narasimham 1, Sean O'Riordan 2,3, Michael Hutchinson 2,3, Richard B Reilly 1,5
1School of Engineering, Trinity College Dublin, The University of Dublin, 2Department of Neurology, St. Vincent's University Hospital, 3School of Medicine and Medical Sciences, University College Dublin, 4School of Mathematical Sciences, Dublin Institute of Technology, 5School of Medicine Trinity College Dublin, The University of Dublin

Methods for the measurement and analysis of the temporal discrimination threshold are presented, and its application to the study of the pathogenesis of cervical dystonia are discussed.

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Immunology and Infection

Dissecting Multi-protein Signaling Complexes by Bimolecular Complementation Affinity Purification (BiCAP)
Jordan F. Hastings 1, Jeremy Z.R. Han 1, Robert F. Shearer 1,2, Sean P. Kennedy 1,3, Mary Iconomou 1,4, Darren N. Saunders 5, David R. Croucher 1,6,7
1The Kinghorn Cancer Centre, Garvan Institute of Medical Research, 2Ubiquitin Signaling Group, Protein Signaling Program, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, 3RCSI Molecular Medicine, Royal College of Surgeons in Ireland, 4Department of Epigenetics, Max Planck Institute of Immunobiology and Epigenetics, 5School of Medical Sciences, University of New South Wales, 6St Vincent's Hospital Clinical School, University of New South Wales, 7School of Medicine and Medical Science, University College Dublin

This manuscript describes the protocol for Bimolecular Complementation Affinity Purification (BiCAP). This novel method facilitates the specific isolation and downstream proteomic characterization of any two interacting proteins, while excluding un-complexed individual proteins as well as complexes formed with competing binding partners.

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Biology

Tissue Collection of Bats for -Omics Analyses and Primary Cell Culture
Laurel R. Yohe *1,2, Paolo Devanna *3, Kalina T.J. Davies 4, Joshua H.T. Potter 4, Stephen J. Rossiter 4, Emma C. Teeling 5, Sonja C. Vernes *3,6, Liliana M. Dávalos *2,7
1Department of Geology & Geophysics, Yale University, 2Department of Ecology & Evolution, Stony Brook University, 3Neurogenetics of Vocal Communication, Max Planck Institute for Psycholinguistics, 4School of Biological and Chemical Sciences, Queen Mary University of London, 5School of Biology & Environmental Science, University College Dublin, 6Donders Institute for Brain, Cognition and Behavior, 7Consortium for Inter-Disciplinary Environmental Research, Stony Brook University

This is a protocol for the optimal tissue preparation for genomic, transcriptomic, and proteomic analyses of bats caught in the wild. It includes protocols for bat capture and dissection, tissue preservation, and cell culturing of bat tissue.

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Biology

Laser Capture Microdissection on Surgical Tissues to Identify Aberrant Gene Expression in Impaired Wound Healing in Type 2 Diabetes
Rachael Williams *1, Irene Castellano-Pelicena *1,2, Aaiad H.A Al-Rikabi 1,3, Stephen K. Sikkink 1, Richard Baker 1, Kirsten Riches-Suman 1, M Julie Thornton 1
1Centre for Skin Sciences, University of Bradford, 2The Charles Institute of Dermatology, School of Medicine, University College Dublin, 3University of Baghdad

This technique provides a guide to inflicting ex vivo wounds, performing laser capture microdissection and quantifying changes in gene expression related to poor wound healing processes in diabetes using clinically relevant human tissue.

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Environment

Sampling, Identification and Characterization of Microplastics Release from Polypropylene Baby Feeding Bottle during Daily Use
Dunzhu Li *1,2, Luming Yang *1,2, Rachel Kavanagh 1, Liwen Xiao 2,3, Yunhong Shi 1,2, Daniel K. Kehoe 1, Emmet D. Sheerin 1,4, Yurii K. Gun’ko 4,5, John J. Boland 1,4, Jing Jing Wang 1
1AMBER Research Centre and Centre for Research on Adaptive Nanostructures and Nanodevices (CRANN), Trinity College Dublin, 2Department of Civil, Structural and Environmental Engineering, Trinity College Dublin, 3TrinityHaus, Trinity College Dublin, 4School of Chemistry, Trinity College Dublin, 5BEACON, Bioeconomy SFI Research Centre, University College Dublin

This study detailed a reliable and cost-effective protocol for microplastics collection and detection from the daily use of plastic products.

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Biology

A Robust Method for the Large-Scale Production of Spheroids for High-Content Screening and Analysis Applications
Alannah S. Chalkley 1, Margaritha M. Mysior 1, Jeremy C. Simpson 1
1Cell Screening Laboratory, UCD School of Biology and Environmental Science, University College Dublin

This protocol details a method for the production of three different types of spheroids in a manner that makes them suitable for large-scale high-content screening and analysis. In addition, examples are presented showing how they can be analyzed at spheroid and individual cell levels.

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Bioengineering

Rapid Antibody Glycoengineering in Chinese Hamster Ovary Cells
Masue Marbiah *1,2, Pavlos Kotidis *1,3, Roberto Donini 1,4, Itzcóatl A. Gómez 5, Ioscani Jimenez del Val 5, Stuart M. Haslam 4, Karen M. Polizzi 1,2, Cleo Kontoravdi 1
1Department of Chemical Engineering, Imperial College London, 2Imperial College Centre for Synthetic Biology, Imperial College London, 3BioPharm Process Research, GlaxoSmithKline Research and Development, 4Department of Life Science, Imperial College London, 5School of Chemical & Bioprocess Engineering, University College Dublin

The glycosylation pattern of an antibody determines its clinical performance, thus industrial and academic efforts to control glycosylation persist. Since typical glycoengineering campaigns are time- and labor-intensive, the generation of a rapid protocol to characterize the impact of glycosylation genes using transient silencing would prove useful.

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Biology

Quantitative Determination of De Novo Fatty Acid Synthesis in Brown Adipose Tissue Using Deuterium Oxide
Rory Turner *1, Rajib Mukherjee *2, Martina Wallace 1, Joan Sanchez-Gurmaches 2,3,4
1UCD Conway Institute and UCD Institute of Food and Health, School of Agriculture and Food Science, University College Dublin, 2Division of Endocrinology, Cincinnati Children’s Hospital Medical Center, Cincinnati, 3Division of Developmental Biology, Cincinnati Children’s Hospital Medical Center, Cincinnati, 4Department of Pediatrics, University of Cincinnati College of Medicine

Here, we present an inexpensive quantitative method utilizing deuterium oxide and gas chromatography mass spectrometry (GCMS) for the analysis of total fatty acid de novo lipogenesis in brown adipose tissue in vivo.

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