Electron microscopic localization of cytoplasmic myosin with ferritin-labeled antibodies.
Population
Vertebrate nonmuscle cells and purified platelet myosin filaments
Comparison
Ferritin-labeled antimyosin antibodies vs Unlabeled antimyosin (preincubation block)
Design
Preclinical
Authors
Discussion
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Member takes
Implication
Offers no immediate clinical guidance; extends ultrastructural mapping of myosin in nonmuscle cells.
Key Points
- To determine the ultrastructural localization and precise subcellular organization of myosin in vertebrate nonmuscle cells using ferritin-conjugated antibodies.
- Purified antibodies against myosin were coupled to ferritin and applied to purified platelet myosin filaments, crude actomyosin preparations, HeLa cells, and mouse intestinal epithelial cells.
- Conjugate specificity was assessed using competitive blocking with unlabeled antimyosin and compared across light immunofluorescence and electron microscopy.
- Ferritin-antimyosin bound selectively to 10–15 nm filaments, with conjugate density along HeLa cell stress fiber dense regions reaching 5- to 6-fold that of intervening regions, 20-fold that of cytoplasm, and 100-fold that of nuclei.
- In mouse intestinal epithelial cells and brush borders exposed to 5 mM MgCl2, ferritin-antimyosin concentrated almost exclusively in the terminal web along 10–15 nm filaments.
Structured PICO
P
Population
Vertebrate nonmuscle cells (HeLa cells, mouse intestinal epithelial cells) and purified platelet myosin filaments
I
Intervention
Ferritin-labeled antimyosin antibodies
C
Comparator
Unlabeled antimyosin (preincubation block)
O
Outcome
Localization of myosin by electron microscopysurrogate
Ferritin-labeled antibodies successfully localized cytoplasmic myosin to specific filament structures and stress fibers in nonmuscle cells using electron microscopy.
Cite This Study
Herman et al. (1981) studied this question.