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Correlative Light and Electron Microscopy for the Investigation of Muscle Disease

Version 4 2016-01-12, 23:50
Version 3 2015-07-24, 05:03
Version 2 2015-07-24, 05:00
Version 1 2015-06-30, 01:14
posted on 2016-01-12, 23:50 authored by Robert Bryson-Richardson, Georg Ramm, Tamar Sztal
In order to investigate the biology of nemaline myopathy, a muscle disease characterised by the formation of nemaline (rod-like) aggregates, we created a zebrafish model.  To generate the model we expressed a disease causing form of ACTA1 (ACTA1D286G) tagged with enhanced green fluorescent protein within the muscle. This allowed the visualisation of disease onset and progression in the living animal. This led to the discovery of how and where the characteristic aggregates form. In order to confirm that the fluorescent aggregates observed in the fish corresponded to those observed in patients using electron microscopy we carried out correlative light and electron microscopy on the zebrafish disease model. This allowed visualisation of both the fluorescent protein and the electron dense aggregates. The raw data from these experiments are provided.