Astroglial-axonal interactions during early stages of myelination in mixed cultures using in vitro and ex vivo imaging techniques
1 Institute of Infection, Immunity and inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Sir Graeme Davies Building, 120 University Place, Glasgow G12 8TA, UK
2 CRUK Beatson Institute, Garscube Estate, Glasgow G61 1BD, UK
BMC Neuroscience 2014, 15:59 doi:10.1186/1471-2202-15-59Published: 2 May 2014
Time-lapse sequence over 21 hours of a dynamic fGFP labelled process from flat astrocyte-like cells in association with a neurite/axon- like structure. A flat fGFP tagged cell process can be seen to dynamically move over a neurite and form a large membranous bleb-like structure of around 10μm in diameter. Additional membranous blebs can be seen to form over time indicating an active motile, membrane.
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Time-lapse imaging of membrane protrusions of fGFP-positive cells in shiverer myelinating cultures after 21 DIV. Time-lapse imaging was carried out for 24 hours with 3 min intervals. Membrane protrusions could be seen to form which changed shape and appeared to be eventually deposited on the axonal surface.
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Ex vivo time-lapse of cGFP-expressing cells typical of astrocytes revealed their dynamic behaviour. cGFP expressing neurospheres were transplanted into the cord 12 days previously. The non-fixed cord was removed and visualised as ex vivo tissue by time lapse over a short time frame. A cGFP tagged astrocyte-like cell could be seen to change the pattern of GFP localisation which suggested a dynamic spontaneous extension of a cell process.
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In vivocGFP-labelled neurospheres were transplanted and a cGFP-labelled cell with long processes resembling an astrocyte was visualised in vivo over a short time frame using time-lapse. Although the movement of breathing could be seen the GFP tagged transplanted cell was clearly visible.
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