Feramisco, J. R. (1979) Microinjection of fluorescently labeled alpha-actinin into living fibroblasts. Proceedings of the National Academy of Sciences of the United States of America, 76 (8). pp. 3967-3971. ISSN 0027-8424
Abstract
alpha-Actinin from chicken gizzard labeled with tetramethylrhodamine isothiocyanate has been incorporated into living fibroblast cells by microinjection. Fluorescent labeling of alpha-actinin was carried out such that the conjugated protein was functional in vitro as shown by its ability to bind to F-actin. Within 1-2 hr after injection, diffuse fluorescence was observed throughout the cytoplasm and only faint fluorescence was apparently associated with the stress fibers. During the ensuing 2-15 hr, however, most of the fluorescence was seen as periodicities along the stress fibers and as foci of the microfilament polygonal networks. This distribution of alpha-actinin in the living cells was strikingly similar to that found by indirect immunofluorescence localization of endogenous alpha-actinin in fixed samples of the same cell type. Control studies in which heat-treated (100 degrees C, 2 min) fluorescent alpha-actinin or tetramethylrhodamine isothiocyanate alone was injected into the cells indicated that the stress fiber and polygonal network labeling was specific for "native" fluorescently labeled alpha-actinin. These results suggest that the dynamic properties of proteins and structures in cultured mammalian cells can be studied with the use of microinjection and fluorescence microscopic techniques.
| Item Type: | Paper |
|---|---|
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > cytoskeletal proteins organs, tissues, organelles, cell types and functions > cell types and functions > cell types > fibroblasts organs, tissues, organelles, cell types and functions > cell types and functions > cell types > fibroblasts organs, tissues, organelles, cell types and functions > cell types and functions > cell types > fibroblasts |
| CSHL Authors: | |
| Communities: | CSHL labs |
| Depositing User: | Matt Covey |
| Date: | 1979 |
| Date Deposited: | 02 Jun 2016 18:54 |
| Last Modified: | 02 Jun 2016 18:54 |
| PMCID: | PMC383957 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/32676 |
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