3 results
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2. Characterization of intermediate filaments and their structural organization during epithelium formation in pigmented epithelial cells of the retina in vitro
- Author
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Hiroaki Sugino, Katsushi Owaribe, and Hirohisa Masuda
- Subjects
Paper ,Histology ,Guinea Pigs ,Immunocytochemistry ,Intermediate Filaments ,Fluorescent Antibody Technique ,Vimentin ,Chick Embryo ,Microfilament ,Retina ,Pathology and Forensic Medicine ,Adherens junction ,medicine ,Animals ,Intermediate Filament Protein ,Pigment Epithelium of Eye ,Intermediate filament ,Cytoskeleton ,Cells, Cultured ,biology ,Demecolcine ,Collodion ,Cell Differentiation ,Cell Biology ,Actins ,Cell biology ,Actin Cytoskeleton ,Microscopy, Electron ,medicine.anatomical_structure ,biology.protein ,Electrophoresis, Polyacrylamide Gel ,Rabbits ,sense organs - Abstract
Retinal pigmented epithelial cells of chicken have circumferential microfilament bundles (CMBs) at the zonula adherens region. Isolated CMBs are polygons filled with a meshwork composed primarily of intermediate filaments; they show three major components of 200 000, 55 000, and 42 000 daltons in SDS-gel electrophoresis. Here we have characterized the 55 000-dalton protein immunochemically and ultrastructurally. Immunoblotting and immunofluorescence microscopy have shown that the 55 000-dalton protein is an intermediate filament protein, vimentin. Vimentin filaments changed their distribution during differentiation of pigmented epithelial cells in culture. The protein in the elongated cells showed a fibroblast-type pattern of intermediate filaments. During epithelium formation, the filaments were uniformly distributed and formed a finer meshwork at the apical level. In pigmented epithelial cells that differentiated and matured in culture, vimentin and actin exhibited their characteristic behavior after treatment with colcemid. In the central to basal region of the cell, intermediate filaments formed thick perinuclear bundles. In the apical region, however, intermediate filaments changed in organization from a nonpolarized meshwork to a polarized bundle-like structure. Simultaneously, new actin bundles were formed, running parallel to the intermediate filaments. This suggests that there is some interaction between microfilaments and intermediate filaments in the apical region of these cells.
- Published
- 1986
3. Immunocytochemical localization of a rhodopsin-like protein in the lipochondria in photosensitive neurons of Aplysia californica
- Author
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Edward O. Anderson, John W. Breneman, Laura J. Robles, Virginia A. Nottoli, and Lori L. Kegler
- Subjects
Central Nervous System ,Paper ,Rhodopsin ,Pathology ,medicine.medical_specialty ,Opsin ,Histology ,Immunocytochemistry ,Fluorescent Antibody Technique ,Immunofluorescence ,Antibodies ,Pathology and Forensic Medicine ,Immunoenzyme Techniques ,Antibody Specificity ,Immunoblot Analysis ,Aplysia ,medicine ,Animals ,Eye Proteins ,Inclusion Bodies ,Neurons ,biology ,medicine.diagnostic_test ,Histocytochemistry ,Decapodiformes ,Rod Opsins ,Collodion ,Cell Biology ,biology.organism_classification ,Lipids ,Molecular biology ,Microscopy, Electron ,medicine.anatomical_structure ,nervous system ,Polyclonal antibodies ,biology.protein ,Electrophoresis, Polyacrylamide Gel ,Neuron ,Retinal Pigments - Abstract
Polyclonal antibodies directed against squid opsin were used in immunocytochemical and immunoblot experiments to identify a rhodopsin-like protein in photosensitive neurons of Aplysia. Aldehyde-fixed abdominal and cerebral ganglia were embedded in paraffin for peroxidase anti-peroxidase analysis or used whole for immunofluorescence studies. Ganglia were embedded in Lowicryl K4M for electron-microscope immunocytochemistry. In both the cerebral and abdominal ganglia, light-microscope immunocytochemical results showed reaction product deposited around the neuronal cell periphery corresponding in position to the lipochondria. In the abdominal ganglion, the giant cell R2, located in the right rostral quarter, and neurons in the right caudal quarter were consistently labeled with anti-opsin. Electron-microscopic studies demonstrated ferritin-labeling of the lipochondria in R2 and other immunoreactive neurons. Immunoblot analysis of R2 and cerebral neuron extracts was used to identify two prominent immunoreactive protein bands at 85 000 and 67 500 molecular weight.
- Published
- 1986
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