Identification and Characterization of Epithelial Cells in Mammalian Tissues by Immunofluorescence Microscopy Using Antibodies to Prekeratin

Franke, Werner W.; Appelhans, B.; Schmid, Erika Nardon; Freudenstein, Christa; Osborn, Mary; Weber, Klaus

doi:10.1111/j.1432-0436.1979.tb01030.x

Cited by 330 publications

(124 citation statements)

References 45 publications

Supporting

Mentioning

111

Contrasting

Unclassified

Order By: Relevance

“…Ofparticular relevance to this report are the keratins that are polypeptide components of the latter two groups. A number of reports have shown the specificity of the keratins not only to epidermis but to nonkeratizing epithelial cells as well (14,(16)(17)(18)(19)(20)(21)(22).The solubility properties ofthe keratins (23-26) are very similar to those we observed for the Novikoff hepatoma nuclear antigens during our purification attempts with these proteins. Moreover, the molecular weights ofthe denatured antigens are very similar to those reported for keratins or prekeratins from a number ofsources (14,15,(26)(27)(28)(29).…”

supporting

confidence: 75%

“…In contrast, the affinity-purified antibodies reacted minimally if at all with Pt K2 cells and with an identical photographic exposure time no immunofluorescence was detected (Fig. 3D) (12)(13)(14)(16)(17)(18)(19)(20)(21)(22).…”

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Identification of cytokeratin antigens in Novikoff ascites hepatoma.

Schmidt¹,

Pardue²,

Tutt³

et al. 1982

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Further characterization of three polypeptide antigens present in Novikoff ascites hepatoma (molecular weights 39,000, 49,000, and 56,000-p39 p49, and p56, respectively), has established the identity of these proteins as cytokeratins. Two-dimensional gel electrophoresis revealed that each of the three proteins exists in several isoelectric forms, all characteristic of the keratins. The antigens p39 and p56 had amino acid compositions compatible with this family of proteins, and immunofluorescence microscopy localized the antigens on elaborate filament arrays indicative of the cytokeratins. Antibodies monospecific for the p39 antigen, purified by using affinity chromatography, were used to demonstrate that protein p39 was specific for Novikoff hepatoma cells. These results suggest that the cytokeratins contain cell-specific species that can be useful as markers for differentiation and neoplasia. (39,000,49,000, and 56,000-p39, p49, and p56, respectively) was originally purified from chromatin (3) and, in contrast to the higher molecular weight group, was also found in cytoplasmic fractions from Novikoff cells (4).The cytoskeletal fiber elements of most eukaryotic cells consist of the microtubules, microfilaments, and intermediatesized filaments. The latter system of filaments is collectively referred to as "intermediate filaments" because their size (6-11 nm) is between the thin microfilaments (5-6 nm) and the thick microtubules (20-25 nm). Examples ofthis category include the neurofilaments and the glial filaments, the 10-nm filaments of muscle, the keratin or prekeratin filaments of keratizing epithelia, and the desmosome-associated tonofilaments of most epithelial cells (5-15). Ofparticular relevance to this report are the keratins that are polypeptide components of the latter two groups. A number of reports have shown the specificity of the keratins not only to epidermis but to nonkeratizing epithelial cells as well (14,(16)(17)(18)(19)(20)(21)(22).The solubility properties ofthe keratins (23-26) are very similar to those we observed for the Novikoff hepatoma nuclear antigens during our purification attempts with these proteins. Moreover, the molecular weights ofthe denatured antigens are very similar to those reported for keratins or prekeratins from a number ofsources (14,15,(26)(27)(28)(29). In this report we have tentatively identified the Novikoff antigens p39, p49, and p56 as members ofthe family ofkeratins, on the basis oftheir preparative properties, isoelectric points, and amino acid compositions. We have used immunofluorescence microscopy to confirm their intracellular location on keratin-like intermediate filaments. Furthermore, the immunological specificity for at least thep39 antigen suggests that this polypeptide may offer distinct possibilities as a probe for neoplasia. MATERIALS AND METHODSNovikoff ascites hepatoma was maintained by weekly transplantations in male Sprague-Dawley rats. The hepatoma cells were washed three times with standard saline citrate as described (3) before us...

show abstract

supporting

confidence: 75%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Identification of cytokeratin antigens in Novikoff ascites hepatoma.

Schmidt¹,

Pardue²,

Tutt³

et al. 1982

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…Prekeratin has been demonstrated in the epidermis and in epithelial cells of the respiratory, intestinal, and urinary tract of mammals and other vertebrates (Franke et al 1979a, b). All these epithelia have been shown to be negative for vimentin (Franke et al 1979b;Schmid et al 1979). However, a systematic study of the amniotic epithelium is still missing except for a short note that such cells have been found to be prekeratin positive after one week of culturing in vitro (Alitalo et al 1980).…”

Section: Introductionmentioning

confidence: 99%

Characterization of cells of amniotic fluids by immunological identification of intermediate-sized filaments: Presence of cells of different tissue origin

et al. 1981

View full text Add to dashboard Cite

“…Cytokeratins are found principally in epithelial cells, while vimentin is a mesenchymal cell type marker (Franke et al, 1979(Franke et al, , 1982. GFAP is used as a marker for fibrous astrocytes (Dahl and Bignami, 1983).…”

mentioning

confidence: 99%

Reorganization of the ependyma during axolotl spinal cord regeneration: Changes in intermediate filament and fibronectin expression

O'Hara¹,

Egar²,

Chernoff³

1992

Developmental Dynamics

View full text Add to dashboard Cite

Changes in intermediate fiiament content and extracellular matrix material showed that the injury response of ependymal cells in lesioned axolotl spinal cord involves an epithelial-to-mesenchymal transformation, and that fibrous astrocytes are excluded from the remodeling lesion site. Antibody localization was used to visualize cytokeratin-, vimentin-, and glial fibrillary acidic protein-(GFAP-) containing intermediate filaments, as well as the adhesive glycoprotein fibronectin. In normal axolotl spinal cord cytokeratins were found near the apical surface of the ependymal cells. Transmission electron microscopic examination suggested that these cytokeratins were in tonofilaments. Cytokeratin expression was lost and vimentin production was initiated in ependymal cells 23 weeks following spinal cord injury. There was a period of approximately 1-2 weeks when cytokeratins and vimentin were co-expressed in vivo. This co-expression was maintained in vitro by culture on a fibronectin-coated substratum. As the central canal reformed, vimentin expression was lost. Ependymal cells lacked GFAP intermediate filaments, but GFAP was present in fibrous astrocytes of the neuropil and white matter. Following injury, GFAP localization showed that fibrous astrocytes disappeared from the remodeling lesion site and reappeared only after the ependymal epithelium reformed and newly myelinated axons were found. Fibronectin expression closely followed the expression of vimentin during mesenchymal ependymal cell outgrowth. These results suggest that the ependymal cell outgrowth requires changes in cell shape followed by changes in production of extracellular matrix.

show abstract

Identification and Characterization of Epithelial Cells in Mammalian Tissues by Immunofluorescence Microscopy Using Antibodies to Prekeratin

Cited by 330 publications

References 45 publications

Identification of cytokeratin antigens in Novikoff ascites hepatoma.

Identification of cytokeratin antigens in Novikoff ascites hepatoma.

Characterization of cells of amniotic fluids by immunological identification of intermediate-sized filaments: Presence of cells of different tissue origin

Reorganization of the ependyma during axolotl spinal cord regeneration: Changes in intermediate filament and fibronectin expression

Contact Info

Product

Resources

About