Scanning Confocal Microscope Detection of Nuclear Actin Using Green Fluorecent Protein (GFP)

17. září 2010 v 14:34 | Cell Biology

„Průkaz jaderného aktinu skenovacím konfokálním mikroskopem pomocí zeleně fluoreskujího proteinu GFP“
(Abstract), in Czech.
P. Kufner, L. Hájková J. Reischig, M. Salášek
Charles Univ. in Praque, Faculty of Medicine in Pilsen, Institute of Biology.

Transgenic Drosophila larvae were studied with respect to the expression and distribution pattern of GFP-coupled proteins. Larval organs were isolated and the fluorescent signal scrutinized using laser scanning confocal microscopy. Cytoplasms, nuclei, and non-cellular backgrounds were submitted to spectral analysis and fluorescence intensity measurements. The cell line expressing GFP-actin exhibited both cytoplasmic and nuclear staining, which was not the case in the remaining two cell lines, whose staining was exclusively cytoplasmic, and which can in this respect be considered as negative controls. The presence of actin in the eukaryotic nucleus has not been unequivocally accepted by the scientific community and the observations remained an enigma until the very recent years when several examples of nuclear functions of actin have been demonstrated. Our findings may represent a significant contribution to the growing understanding of the role of actin in the nucleus.

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