Latest finding - Molecular Cell Biology
- Single molecule microscopy in living zebrafish embryos
Advanced microscopy techniques have been used for some time to visualize individual fluorescently labeled molecules within the plasma membrane of living cells. We extended this technology to the level of a whole living organism (in vivo) using so-called total internal reflection fluorescence (TIRF) microscopy. We injected mRNA encoding yellow fluorescent protein tagged to a membrane anchor into early stage zebrafish embryos, and visualized the fluorescent molecules in the plasma membranes of epidermal cells in the tail region.
This technique exposed the diffusion patterns of individual molecules, revealing two subpopulations of either fast or slow molecules. Similar subpopulations of molecules with different diffusion rates were observed in zebrafish embryonic fibroblast cell lines (in vitro), and in primary stem cell cultures derived from embryos four hours after fertilization (ex vivo). However, we found an important discrepancy: fast molecules were freely diffusing in vivo while they were confined within a domain in vitro and ex vivo.
The observed difference in confinement of the diffusion is probably caused by physical barriers formed by the membrane cytoskeleton in the in vitro and ex vivo conditions. We think that this finding clearly illustrates the relevance of performing this type of studies in an intact living organism. In future studies we will use the in vivo approach to explore the movements of membrane proteins during signaling events, for example during skin cancer, inflammation, wound healing and regeneration.
- Biophysical journal
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Marcel Schaaf et al., published their finding in august 2009 in the Biophysical Journal
- Other news from the MCB group
- EU Funding for Marie-Curie International Training Network: FishForPharma
- LUF subsidy for tuberculosis research
- PlosOne paper on automated injection of zebrafish embryos
- Annemarie Meijer discoverer of the year 2010
- NWO VENI grant for Christian Tudorache
- NWO TOP grant for modeling human cancer in zebrafish
- Blood paper on macrophage-specific genes in zebrafish embryos
