A new look at the cellular scaffold by embedment-free electron microscopy method.
The basic scaffold of most cells is afforded by the cytoskeleton (comprising microfilaments, intermediate filaments and the microtubules). The conventional methods of electron microscopy fail to visualize filamentous cell structure. They can show only these filaments lying at the section surface. Heavy metal staining (I), and the optical properties of the resins used for embedding are similar to those of proteins hence most proteinaceous structures remain unresolved and the cytoplasm seems to be quite homogenous (II). Aldehyde fixation could cross-link proteins and lead to the emergence of artificial structures (III). These limitations may be overcome by the use of the embedment-free electron microscopy (EF-EM). This technique present cellular scaffold as a purified, isolated, three-dimensional network with various thickness of filaments. Our study on the dynamic aspect of cellular scaffold indicate that the thickness and arrangement of filaments depend on cell type and both physiological or pathological environments. Thank also to the adaptation of immunocytochemistry to EF-EM it was possible to understand the nuclear matrix and cytomatrix structure in relation to function. Thus, combination these methods revealed findings suggesting the nuclear homing of proapoptotic proteins and their association with intermediate filaments.
Laboratory of Cell Ultrastructure, Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland. gajk@cmdik.pan.pl
Barbara
B
Gajkowska
Urszula
U
Wojewódzka
D000818:Animals; D051023:BH3 Interacting Domain Death Agonist Protein; D002352:Carrier Proteins / Q000032:analysis; D045744:Cell Line, Tumor; D003599:Cytoskeleton / Q000648:ultrastructure*; D006801:Humans; D007150:Immunohistochemistry; D008854:Microscopy, Electron / Q000379:methods*; D015530:Nuclear Matrix / Q000648:ultrastructure; D011092:Polyethylene Glycols / Q000737:chemistry; D018272:Porins / Q000032:analysis; D011518:Proto-Oncogene Proteins / Q000032:analysis; D019253:Proto-Oncogene Proteins c-bcl-2; D016707:Tissue Fixation / Q000379:methods*; D050994:Voltage-Dependent Anion Channels; D051028:bcl-2-Associated X Protein
D051023:BH3 Interacting Domain Death Agonist Protein; C494814:BID protein, human; D002352:Carrier Proteins; D018272:Porins; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D050994:Voltage-Dependent Anion Channels; D051028:bcl-2-Associated X Protein; D011092:Polyethylene Glycols