Projects per year
Abstract
An experimental method has been developed to investigate the cellular responses to central nervous system (CNS) injury using the fruit-fly Drosophila. Understanding repair and regeneration in animals is a key question in biology. The damaged human CNS does not regenerate, and understanding how to promote regeneration is one of main goals of medical neuroscience. The powerful genetic toolkit of Drosophila can be used to tackle the problem of CNS regeneration.
A lesion to the CNS ventral nerve cord (VNC, equivalent to the vertebrate spinal cord) is applied manually with a tungsten needle. The VNC can subsequently be filmed in time-lapse using laser scanning confocal microscopy for up to 24 hours to follow the development of the lesion over time. Alternatively, it can be cultured, then fixed and stained using immunofluorescence to visualise neuron and glial cells with confocal microscopy. Using appropriate markers, changes in cell morphology and cell state as a result of injury can be visualized. With ImageJ and purposely developed plug-ins, quantitative and statisical analyses can be carried out to measure changes in wound size over time and the effects of injury in cell proliferation and cell death. These methods allow the analysis of large sample sizes. They can be combined with the powerful genetics of Drosophila to investigate the molecular mechanisms underlying CNS regeneration and repair.
A lesion to the CNS ventral nerve cord (VNC, equivalent to the vertebrate spinal cord) is applied manually with a tungsten needle. The VNC can subsequently be filmed in time-lapse using laser scanning confocal microscopy for up to 24 hours to follow the development of the lesion over time. Alternatively, it can be cultured, then fixed and stained using immunofluorescence to visualise neuron and glial cells with confocal microscopy. Using appropriate markers, changes in cell morphology and cell state as a result of injury can be visualized. With ImageJ and purposely developed plug-ins, quantitative and statisical analyses can be carried out to measure changes in wound size over time and the effects of injury in cell proliferation and cell death. These methods allow the analysis of large sample sizes. They can be combined with the powerful genetics of Drosophila to investigate the molecular mechanisms underlying CNS regeneration and repair.
Original language | English |
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Article number | e50306 |
Journal | Journal of Visualized Experiments |
Volume | 73 |
DOIs | |
Publication status | Published - 28 Mar 2013 |
Keywords
- Drosophila
- repair
- CNS
- regeneration
- Repo
- glia
- larva
ASJC Scopus subject areas
- General Neuroscience
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Dive into the research topics of 'An Injury paradigm to investigate central nervous system repair in Drosophila'. Together they form a unique fingerprint.Projects
- 2 Finished
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The molecular control of glial progenitor proliferation in Drosophila and mammals
Hidalgo, A. (Principal Investigator) & Logan, A. (Co-Investigator)
Biotechnology & Biological Sciences Research Council
4/01/10 → 30/06/13
Project: Research Councils
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Confocal Microscopy for Time-lapse, FRET and Calcium Imaging Analysis of Cellular Processes
Hidalgo, A. (Principal Investigator), Hotchin, N. (Co-Investigator), Machesky, L. (Co-Investigator) & Publicover, S. (Co-Investigator)
18/10/04 → 17/03/10
Project: Research