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  • The chondrocyte cytoskeleton in mature articular cartilage: structure and distribution of actin, tubulin, and vimentin filaments.

The chondrocyte cytoskeleton in mature articular cartilage: structure and distribution of actin, tubulin, and vimentin filaments.

The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society (2000-09-16)
E Langelier, R Suetterlin, C D Hoemann, U Aebi, M D Buschmann
ABSTRACT

We investigated the structure of the chondrocyte cytoskeleton in intact tissue sections of mature bovine articular cartilage using confocal fluorescence microscopy complemented by protein extraction and immunoblotting analysis. Actin microfilaments were present inside the cell membrane as a predominantly cortical structure. Vimentin and tubulin spanned the cytoplasm from cell to nuclear membrane, the vimentin network appearing finer compared to tubulin. These cytoskeletal structures were present in chondrocytes from all depth zones of the articular cartilage. However, staining intensity varied from zone to zone, usually showing more intense staining for the filament systems at the articular surface compared to the deeper zones. These results obtained on fluorescently labeled sections were also corroborated by protein contents extracted and observed by immunoblotting. The observed cytoskeletal structures are compatible with some of the proposed cellular functions of these systems and support possible microenvironmental regulation of the cytoskeleton, including that due to physical forces from load-bearing, which are known to vary through the depth layers of articular cartilage.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Phalloidin–Tetramethylrhodamine B isothiocyanate, sequence from Amanita phalloides(synthetic: peptide sequence)
Sigma-Aldrich
Monoclonal Anti-Actin antibody produced in mouse, clone AC-40, ascites fluid
Sigma-Aldrich
Monoclonal Anti-Vimentin antibody produced in mouse, clone V9, ascites fluid
Sigma-Aldrich
Glycerol, BioXtra, ≥99% (GC)