Griffith University

3 ARTICLES PUBLISHED IN JoVE

Developmental Biology

Real-time Live-cell Flow Cytometry to Investigate Calcium Influx, Pore Formation, and Phagocytosis by P2X7 Receptors in Adult Neural Progenitor Cells

Hannah C. Leeson^1,2, Tailoi Chan-Ling^3,4, Michael D. Lovelace^3,5,6, Jeremy D. Brownlie⁷, Michael W. Weible II^*1,4,7, Ben J. Gu^*8

¹Griffith Institute for Drug Discovery, Griffith University, ²Australian Institute for Bioengineering and Nanotechnology, University of Queensland, ³Discipline of Anatomy and Histology, School of Medical Science, University of Sydney, ⁴Bosch Institute, University of Sydney, ⁵Applied Neurosciences Program, Peter Duncan Neurosciences Research Unit, St. Vincent's Centre for Applied Medical Research, ⁶School of Medical Sciences, The University of New South Wales (UNSW) Medicine, Sydney, New South Wales, ⁷School of Environment and Science, Griffith University, Brisbane, Queensland, ⁸Florey Institute of Neuroscience and Mental Health, University of Melbourne

Providing single-cell sensitivity, real-time flow cytometry is uniquely suited to quantify multimodal receptor functions of live cultures. Using adult neural progenitor cells, the P2X7 receptor function was assessed via calcium influx detected by calcium indicator dye, transmembrane pore formation by ethidium bromide uptake, and phagocytosis using fluorescent latex beads.

Medicine

Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology

Michael A. Pearen^*1, Hong Kiat Lim^*1, Francis D. Gratte^2,3, Manuel A. Fernandez-Rojo^1,4,5, Sujeevi K. Nawaratna⁶, Geoffrey N. Gobert⁷, John K. Olynyk^8,9, Janina E. E. Tirnitz-Parker^2,10, Grant A. Ramm^1,4

¹Hepatic Fibrosis Group, QIMR Berghofer Medical Research Institute, ²School of Pharmacy and Biomedical Sciences, Curtin Health Innovation Research Institute, Curtin University, ³School of Veterinary and Life Sciences, Murdoch University, ⁴School of Medicine, The University of Queensland, ⁵Madrid Institute for Advanced Studies (IMDEA) in Food, CEI UAM+CSIC, ⁶School of Medicine, Griffith University, ⁷School of Biological Sciences, Queen's University Belfast, ⁸Department of Gastroenterology & Hepatology, Fiona Stanley and Fremantle Hospital Group, ⁹School of Medical and Health Sciences, Edith Cowan University, ¹⁰Centre for Cell Therapy and Regenerative Medicine, School of Biomedical Sciences, University of Western Australia

This protocol provides a simple and reliable method for the production of viable precision-cut liver slices from mice. The ex vivo tissue samples can be maintained under laboratory tissue culture conditions for multiple days, providing a flexible model to examine liver pathobiology.

Neuroscience

Induction of Complete Transection-Type Spinal Cord Injury in Mice

Ronak Reshamwala^1,2,3, Tanja Eindorf^2,3, Megha Shah^2,3, Graham Smyth^2,3, Todd Shelper^2,3, James St. John^*1,2,3, Jenny Ekberg^*2,3

¹Griffith Institute for Drug Discovery, Griffith University, ²Menzies Health Institute Queensland, Griffith University, ³Clem Jones Centre for Neurobiology and Stem Cell Research, Griffith University

This protocol describes how to create a precise laminectomy for induction of stable transection-type spinal cord injury in the mouse model, with minimal collateral damage for spinal cord injury research.