JoVE Logo

Sign In

University of British Columbia

10 ARTICLES PUBLISHED IN JoVE

image

Medicine

A Contusive Model of Unilateral Cervical Spinal Cord Injury Using the Infinite Horizon Impactor
Jae H.T. Lee 1, Femke Streijger 1, Seth Tigchelaar 1, Michael Maloon 1, Jie Liu 1, Wolfram Tetzlaff 1, Brian K. Kwon 1,2
1International Collaboration on Repair Discoveries (ICORD), University of British Columbia , 2Department of Orthopaedics, University of British Columbia

A reliable and repeatable way to produce a cervical unilateral spinal cord injury using the Infinite Horizon impactor is described. The method takes advantage of a custom designed frame and clamp to stabilize the spine. The standardized procedure and biomechanical injury parameters result in sufficient and sustained injuries.

image

Biology

A Step-by-step Method for the Reconstitution of an ABC Transporter into Nanodisc Lipid Particles
Huan Bao 1, Franck Duong 1, Catherine S. Chan 1
1Department of Biochemistry and Molecular Biology, University of British Columbia

Nanodiscs are small discoid particles that incorporate membrane proteins into a small patch of phospholipid bilayer. We provide a visual protocol that shows the step-by-step incorporation of the MalFGK2 transporter into a disc.

image

Biology

Measuring Cell Cycle Progression Kinetics with Metabolic Labeling and Flow Cytometry
Helen Fleisig 1, Judy Wong 1
1Faculty of Pharmaceutical Sciences, University of British Columbia

Tracking subtle changes in the progression and kinetics of cell cycle stages can be accomplished by use of a combination of metabolic labeling of nucleic acids with BrdU and total genomic DNA staining via Propidium Iodide. This method avoids the need of chemical synchronization of cycling cells, thereby preventing the introduction of non-specific DNA damage, which in turn affects cell cycle progression.

image

Immunology and Infection

Depletion and Reconstitution of Macrophages in Mice
Shelley B. Weisser 1, Nico van Rooijen 2, Laura M. Sly 3
1Department of Graduate Studies, University of British Columbia , 2Department of Molecular Biology, Vrije Universiteit Amsterdam, 3Department of Pediatrics, University of British Columbia

Macrophages play a central role in homeostasis and pathology in many tissues. The protocol presented here describes methods for depleting macrophages in vivo, deriving polarized macrophages from bone marrow aspirates, and adoptively transferring macrophages into mice. These techniques allow determination of the role that polarized macrophages play in health and disease.

image

Neuroscience

Detection of Protein Palmitoylation in Cultured Hippocampal Neurons by Immunoprecipitation and Acyl-Biotin Exchange (ABE)
G. Stefano Brigidi 1, Shernaz X Bamji 1
1Department of Cellular and Physiological Sciences, Brain Research Centre, University of British Columbia

The reversible addition of palmitate to proteins is an important regulator of intracellular protein trafficking. This is of particular interest in neurons where many synaptic proteins are palmitoylated. We utilize a simple biochemical method to detect palmitoylated proteins in cultured neurons, which can be adapted for multiple cell types and tissues.

image

Immunology and Infection

Antigens Protected Functional Red Blood Cells By The Membrane Grafting Of Compact Hyperbranched Polyglycerols
Rafi Chapanian 1,2, Iren Constantinescu 1,3, Donald E. Brooks 1,2,4, Mark D. Scott 1,2,3, Jayachandran Kizhakkedathu 1,2,4
1Centre for Blood Research, University of British Columbia , 2Department of Pathology and Laboratory Medicine, University of British Columbia , 3Canadian Blood Services, University of British Columbia , 4Department of Chemistry, Life Sciences Centre, University of British Columbia

The cell membrane modification of red blood cells (RBCs) with hyperbranched polyglycerol (HPG) is presented. Modified RBCs were characterized by aqueous two phase partitioning, osmotic fragility and complement mediated lysis. The camouflage of surface proteins and antigens was evaluated using the flow cytometry and Micro Typing System (MTS) blood phenotyping cards.

image

Medicine

Movement Retraining using Real-time Feedback of Performance
Michael Anthony Hunt 1
1Department of Physical Therapy, University of British Columbia

Retraining abnormal movement patterns following injury or disease is a key component of physical rehabilitation. Recent advances in technology have permitted accurate assessment of movement during a variety of tasks, with near instantaneous quantification of results. This provides new opportunities for modification of faulty movement patterns in real time.

image

Immunology and Infection

Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species
Billie Velapatiño 1, James E. A. Zlosnik 1, Trevor J. Hird 1, David P. Speert 1
1Department of Pediatrics, Centre for Understanding and Preventing Infection in Children, University of British Columbia

Members of the Burkholderia genus are pathogens of clinical importance. We describe a method for total bacterial protein extraction, using mechanical disruption, and 2-D gel electrophoresis for subsequent proteomic analysis.

image

Immunology and Infection

Assessment of Antibody-based Drugs Effects on Murine Bone Marrow and Peritoneal Macrophage Activation
Lisa Kozicky 1, Laura M. Sly 1
1British Columbia Children's Hospital Research Institute, University of British Columbia

Antibody-based drugs have revolutionized treatment for inflammatory diseases. In addition to having direct effects on specific targets, antibodies can activate macrophages to become anti-inflammatory. This protocol describes how anti-inflammatory macrophage activation can be assessed in vitro, using mouse bone marrow macrophages, and in vivo, using peritoneal macrophages.

image

Biology

Nucleofection and In Vivo Propagation of Chicken Eimeria Parasites
Chunhui Duan 1, Xinming Tang 1, Dandan Hu 1, Sixin Zhang 1, Jie Liu 1, Feifei Bi 1, Zhenkai Hao 1, Jingxia Suo 1, Ying Yu 1, Min Wang 1, Pei Sun 1, Liying Du 2, Xun Suo 1, Xianyong Liu 1
1National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, 2School of Life Sciences, Peking University

Here, we provided a method to achieve stable transfection of chicken Eimeria parasites by nucleofecting sporozoites or second-generation merozoites. Genetically modified eimerian parasites expressing heterologous antigenic genes could be used as vaccine delivery vehicles.

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved