Coalignment of microtubules, cytokeratin intermediate filaments, and collagen fibrils in a collagen-secreting cell system.
The distribution of microtubules and intermediate filaments in the collagen-secreting scleroblasts of the goldfish scale was investigated by immunofluorescence and electron microscopy. Many of the microtubules and cytokeratin type intermediate filaments formed bundles that were aligned with the underlying, parallel collagen fibrils. The intermediate filament bundles were evenly spaced and located adjacent to the basal plasma membrane. The microtubules, on the other hand, were located further away from the membrane, although many were found very close to the intermediate filament bundles. No detectable change was observed in scleroblast microtubules when cells on scales were treated with colchicine or cooled (greater than or equal to 0 degrees C) for up to 1 h. Cells had to be cooled overnight before the microtubules were affected. The final number and length of the microtubules in the cell depended only on the final steady-state temperature and not the temperature history of the scale cell, and steady state was reached more slowly at colder temperatures. The microtubules but not the intermediate filaments rapidly (within 5 min) and reversibly depolymerized when cells were chilled to -2 approximately -4 degrees C. When chilled cells were warmed, the microtubules polymerized back, within 15 min at room temperature, to the same pattern of parallel coalignment with the underlying collagen. They appeared to repolymerize via two different pathways: (1) a radial growth outwards from the microtubule organizing center followed by a progressive realignment with the underlying collagen and (2) a gradual and simultaneous polymerization along cold-stable, antitubulin staining fibers. These fibers were also aligned with the collagen fibrils and may be related to the aligned intermediate filaments.
McBeath
E
E
Department of Structural Analysis, National Cardiovascular Center Research Institute, Osaka, Japan.
Fujiwara
K
K
D000818:Animals; D003078:Colchicine / Q000494:pharmacology; D003094:Collagen / Q000378:metabolism*; D003238:Connective Tissue / Q000378:metabolism / Q000648:ultrastructure*; D003599:Cytoskeleton / Q000648:ultrastructure*; D006054:Goldfish; D007150:Immunohistochemistry; D007382:Intermediate Filaments / Q000648:ultrastructure*; D007633:Keratins / Q000032:analysis*; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008870:Microtubules / Q000187:drug effects / Q000648:ultrastructure*; D015739:Nocodazole / Q000494:pharmacology; D013696:Temperature
D046911:Macromolecular Substances; D007633:Keratins; D003094:Collagen; D015739:Nocodazole; D003078:Colchicine