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PubMed1995

Expression of desmin and vimentin intermediate filaments in human decidual cells during first trimester pregnancy.

Human endometrial stromal cells (decidual cells) display dramatic alterations in cell shape and size during decidualization. The present study was designed to demonstrate the expression of two major cytoskeletal elements, desmin and vimentin, in human pregnant endometrial decidual cells. Additionally, stage-dependent variations of those intermediate filaments (IFs) among gestational weeks were also evaluated with regard to the support and maintenance of decidualization. Materials were obtained from legal suction terminations of pregnancies of 3-10 weeks gestation. Tissue specimens were either blocked in paraffin or enzymatically dissociated for isolation of decidual cells which subsequently were cultured as monolayers. Immunoperoxidase and immunofluorescence staining methods were applied by using anti-desmin and anti-vimentin monoclonal antibodies. Both desmin and vimentin expression were observed during the early weeks of pregnancy (3-6 weeks). These two types of IFs were also detected in short-term cultures in a filamentous fashion either within the cell body or at cellular attachment plaques. When decidual cells were cultured for longer periods (40-60 days), the expression of desmin dramatically declined while vimentin expression was maintained in a rather diffuse and more abundant fashion. The in situ expression of desmin and vimentin in later weeks of gestation (7-10 weeks) correlated with immunofluorescence staining of long-term cultured cells in that desmin staining was very weak and mostly undetectable where vimentin expression persisted and was evenly distributed throughout the entire stroma. The results demonstrate the differential expression of two major IFs, desmin and vimentin, in human endometrial stromal cells during decidualization and subsequent placentation. The persistence of vimentin in all stages examined suggests that this IF is probably involved in cell morphology and nucleocytoplasmic integrity. The temporal pattern of desmin expression suggests a role for this IF during the rapid onset of the decidualization process.

Cells, Cultured↗
PubMedDate not supplied

[The distribution of intermediate filaments, histiocytic antigens and neural crest-related antigens in the adult dental pulp. An immunocytochemical approach].

MATERIALS AND METHODS: Ten human healthy teeth were extracted for orthodontic or surgical purposes and processed for histological and immunocytochemical examination with the streptavidin-biotin-peroxidase method. Highly purified antibodies were used to reveal the distribution of intermediate filaments (vimentin, desmin, alpha-smooth muscle actin), histiocytic antigens (alpha-1-antitrypsin, lysozyme), neural and neural-crest-associated antigens (neuron-specific enolase, chromogranin-A, S-100, synaptophysin, glial fibrillary acidic protein, neurofilaments) in the adult pulp tissue. RESULTS: Vimentin immunostaining was strongly positive in the network of pulpal fibroblasts. Desmin and alpha-smooth muscle actin were present only in the vessel walls. Staining for alpha-1-antitrypsin and lysozyme reveals a number of macrophage-like cells in the central portion of the pulp. Macrophages were the most dominating immunocompetent cells. Negative immunostaining for chromogranin-A demonstrated the absence of neuroendocrine antigens in the adult dental pulp. Immunostaining for neuron-specific enolase, S-100 protein, synaptophysin, glial fibrillary acidic protein and neurofilaments were positive, with different intensity, in nerve fibres, but no pulp cell was found to be immunoreactive. CONCLUSIONS: The conclusion is drawn that the lack of labelling of pulp cells by the neural associated antibodies could be due to differentiation processes during tissue development.

Adult↗
PubMed1987

The relationship between intermediate filaments and microfilaments before and during the formation of desmosomes and adherens-type junctions in mouse epidermal keratinocytes.

Actin, keratin, vinculin and desmoplakin organization were studied in primary mouse keratinocytes before and during Ca2+-induced cell contact formation. Double-label fluorescence shows that in cells cultured in low Ca2+ medium, keratin-containing intermediate filament bundles (IFB) and desmoplakin-containing spots are both concentrated towards the cell center in a region bounded by a series of concentric microfilament bundles (MFB). Within 5-30 min after raising Ca2+ levels, a discontinuous actin/vinculin-rich, submembranous zone of fluorescence appears at cell-cell interfaces. This zone is usually associated with short, perpendicular MFB, which become wider and longer with time. Later, IFB and the desmoplakin spots are seen aligned along the perpendicular MFB as they become redistributed to cell-cell interfaces where desmosomes form. Ultrastructural analysis confirms that before the Ca2+ switch, IFB and desmosomal components are found predominantly within the perimeter defined by the outermost of the concentric MFB. Individual IF often splay out, becoming interwoven into these MFB in the region of cell-substrate contact. In the first 30 min after the Ca2+ switch, areas of submembranous dense material (identified as adherens junctions), which are associated with the perpendicular MFB, can be seen at newly formed cell-cell contact sites. By 1-2 h, IFB-desmosomal component complexes are aligned with the perpendicular MFB as the complexes become redistributed to cell-cell interfaces. Cytochalasin D treatment causes the redistribution of actin into numerous patches; keratin-containing IFB undergo a concomitant redistribution, forming foci that coincide with the actin-containing aggregates. These results are consistent with an IF-MF association before and during desmosome formation in the primary mouse epidermal keratinocyte culture system, and with the temporal and spatial coordination of desmosome and adherens junction formation.

Actin Cytoskeleton↗
PubMed1988

Maturation of hagfish gland thread cells: composition and characterization of intermediate filament polypeptides.

Previous studies with the hagfish, a primitive vertebrate, have shown that the gland thread cells (GTCs) each contain a single thread (approximately 60 cm long in average-sized cells) in the form of a concisely coiled cytoskeletal entity destined for export by holocrine secretion. The thread in relatively immature GTCs consists almost entirely of intermediate filaments (IFs) bundled in parallel alignment with far fewer microtubules (MTs). The three thread polypeptides described earlier (alpha, basic; beta, acidic; gamma, most acidic; each with a Mr of 63-64 kD) are now further evaluated with respect to in vitro assembly, cross-reactivity with IF polypeptides from higher vertebrates, and peptide sequence homology with known IF polypeptides. The overall results mainly suggest that the hagfish polypeptides are keratinlike substances but lamins or a new type of IF is not ruled out. However, cross-reactivity is weak with mammalian keratins; the 8-11-nm filaments formed from mixtures of alpha and gamma in vitro are generally linear rather than the curvilinear structures usually formed by keratin and nonkeratin IFs; and mixtures of alpha and beta tend to yield 9-12-nm granules or granular strings. Polypeptide analyses on GTCs segregated on the basis of maturational stage show a progressive increase in beta/gamma values which correlates with cell maturation, but the alpha/(beta + gamma) ratios remain near 1. Inasmuch as beta and gamma have many similar properties, the documented increase in the amount of the beta component in aging GTCs might in part be the result of a failure in a posttranslational modification system and may contribute to the ultrastructural changes that accompany thread maturation in preparation for holocrine secretion and subsequent modulation of the viscoelastic properties of mucus.

Animals↗
PubMed1989

Differential diagnosis of genitourinary tumors using monoclonal antibodies to intermediate filament proteins.

Definitive diagnosis of poorly differentiated and metastatic neoplasms may be impossible using conventional histologic criteria. Recent developments in cell biology and immunology now enable us to answer such difficult diagnostic problems. Several varieties of structural proteins can be identified in malignant cells using monoclonal antibodies. The composition of these proteins can yield information regarding the origin of a neoplasm. Intermediate filaments are one such family of structural proteins. By characterization of these proteins, using a panel of monoclonal antibodies, poorly differentiated tumors may be definitively classified as carcinomas, sarcomas, lymphomas, or neural tumors. This approach to tumor diagnosis is now applicable to difficult problems in clinical urology.

Adenocarcinoma↗
PubMed1987

Intermediate filament expression in human vascular smooth muscle and in arteriosclerotic plaques.

Different regions of human aorta and of other human arteries obtained at autopsy were analyzed with regard to their topography and to the different stages of arteriosclerosis. Material was studied by immunocytochemical techniques with antibodies specific for either desmin (D) or for vimentin (V), the two types of intermediate filament proteins present in vascular smooth muscle cells. In normal arteries endothelial cells as well as the adjacent intimal cells were D-V+. In the media D+V+ as well as D-V+ cells were present, with the relative numbers of each cell type dependent on the particular blood vessel. When cells in arteriosclerotic plaques at different stages of development were examined an occasional plaque showed cells of the D+V+ type. In the majority of plaques however the cells were V-D+. In plaques where severe ulceration and necrotic material was present D-V+ cells were found at the border of the lesion: foam cells when they could be identified appeared to be D-V+.

Adult↗
PubMed2000

Development of chick cardiomyocytes: modulation of intermediate filaments by basic fibroblast and platelet-derived growth factors.

Recent studies suggest that peptide growth factors play a functional role in cardiac muscle. To test whether embryonic cardiac muscle is a target for regulation by basic fibroblast growth factor and platelet-derived growth factor, we analyzed the effects of these peptides on the expression of the intermediate filaments desmin and vimentin at the subcellular level during development. Sodium dodecyl sulfate-gel electrophoresis, immunoblotting and fluorescence-activated cell sorting analysis were used to study the effect of basic fibroblast growth factor and platelet-derived growth factor on cultures of chick cardiomyocytes during development. Cytoplasmic and cytoskeletal concentrations of desmin and vimentin were dependent on the stage of embryonic development and on the type of growth factor added to the culture. The most significant finding was the increase in desmin expression in the cytoplasmic and cytoskeletal compartments after treatment with basic fibroblast growth factor (10 ng/ml) of chick heart cells at Hamburger and Hamilton stage 19. In more mature stages, basic fibroblast growth factor did not modify the levels of desmin expression. However, this factor led to a progressive deceleration in the rate of increase in vimentin expression. Platelet-derived growth factor increased vimentin expression in all stages studied, the greatest increases appearing in early stages of heart development. Our findings support the hypothesis that basic fibroblast growth factor plays a role in cardiomyocyte differentiation during the early stages of development, whereas platelet-derived growth factor has a dedifferentiating effect.

Animals↗
PubMed1998

Biologic determinants of uveal melanoma metastatic phenotype: role of intermediate filaments as predictive markers.

The long-range goal of our research is to develop intervention strategies based on newly discovered biologic mechanisms responsible for the invasive dissemination of metastatic uveal melanoma. To accomplish this goal, we have focused on the biologic relevance of novel marker proteins contributing to the uveal melanoma metastatic phenotype. The expression of vimentin intermediate filaments (IFs), a mesenchymal marker, is typical of melanomas, whereas carcinomas typically express keratin IFs, which are markers for epithelia. Thus, cells that coexpress both IFs are regarded as "interconverted" in that they display both mesenchymal and epithelial phenotypes. Although the biologic functions of IFs have remained enigmatic, there is substantial support to suggest that the significance of vimentin/keratin coexpression is linked with poor patient outcome in cutaneous melanoma. Our data demonstrate that human uveal melanoma cell lines (isolated from primary choroidal or ciliary body melanomas and from foci of metastatic uveal melanoma to the liver), which contain predominant populations of cells that coexpress vimentin/keratins 8 and 18 (keratins 8,18) IFs, were 6-fold more invasive through collagenous extracellular matrices in vitro, compared with uveal melanoma cells expressing vimentin only, and were 8- to 13-fold more invasive than normal uveal melanocytes. Colocalization of vimentin/keratins 8,18 in cell cultures was corroborated by immunohistochemistry in histologic sections of tumors from which the cell lines were derived. Minor populations of these cells also coexpressed keratins 13 and 17. Experimental down-regulation of the predominant keratins 8,18 in the interconverted cells, using 16-mer antisense oligonucleotides, resulted in a significant decrease in the migratory ability of the cells-similar to levels achieved by cells positive only for vimentin. These findings provide justification for additional studies of the association between coexpression of IFs vimentin/keratins 8,18 and uveal melanoma metastasis.

Antisense Elements (Genetics)↗
PubMed1984

Epithelial cytoskeletal framework and nuclear matrix-intermediate filament scaffold: three-dimensional organization and protein composition.

Madin-Darby canine kidney (MDCK) cells grow as differentiated, epithelial colonies that display tissue-like organization. We examined the structural elements underlying the colony morphology in situ using three consecutive extractions that produce well-defined fractions for both microscopy and biochemical analysis. First, soluble proteins and phospholipid were removed with Triton X-100 in a physiological buffer. The resulting skeletal framework retained nuclei, dense cytoplasmic filament networks, intercellular junctional complexes, and apical microvillar structures. Scanning electron microscopy showed that the apical cell morphology is largely unaltered by detergent extraction. Residual desmosomes, as can be seen in thin sections, were also well-preserved. The skeletal framework was visualized in three dimensions as an unembedded whole mount that revealed the filament networks that were masked in Epon-embedded thin sections of the same preparation. The topography of cytoskeletal filaments was relatively constant throughout the epithelial sheet, particularly across intercellular borders. This ordering of epithelial skeletal filaments across contiguous cell boundaries was in sharp contrast to the more independent organization of networks in autonomous cells such as fibroblasts. Further extraction removed the proteins of the salt-labile cytoskeleton and the chromatin as separate fractions, and left the nuclear matrix-intermediate filament (NM-IF) scaffold. The NM-IF contained only 5% of total cellular protein, but whole mount transmission electron microscopy and immunofluorescence showed that this scaffold was organized as in the intact epithelium. Immunoblots demonstrate that vimentin, cytokeratins, desmosomal proteins, and a 52,000-mol-wt nuclear matrix protein were found almost exclusively in the NM-IF scaffold. Vimentin was largely perinuclear while the cytokeratins were localized at the cell borders. The 52,000-mol-wt nuclear matrix protein was confined to the chromatin-depleted matrix and the desmosomal proteins were observed in punctate polygonal arrays at intercellular junctions. The filaments of the NM-IF were seen to be interconnected, via the desmosomes, over the entire epithelial colony. The differentiated epithelial morphology was reflected in both the cytoskeletal framework and the NM-IF scaffold.

Animals↗
PubMed1997

Fluorescence microscopic comparison of the binding of phosphodiester and phosphorothioate (antisense) oligodeoxyribonucleotides to subcellular structures, including intermediate filaments, the endoplasmic reticulum, and the nuclear interior.

To detect potential intracellular binding sites for antisense oligodeoxyribonucleotides (ODN), 3'-fluorescence-tagged phosphodiester (P) and phosphorothioate (S) analogs of a series of model and vimentin and actin antisense ODN were applied to digitonin-permeabilized fibroblast and epithelial PtK2 cells. Fluorescence microscopy revealed binding of the ODN to intermediate filaments (IFs) with a preference for cytokeratin IFs, cytoplasmic membranes (endoplasmic reticulum), and, above all, the nuclear interior. The affinity of the ODN for these cellular substructures was dependent on their base composition, and the S-ODN were by far superior to the corresponding P-ODN in binding activity. Fluorescence polarization measurements of the interaction of ODN with purified IF proteins in vitro confirmed the differential, high-affinity binding of S-ODN to IFs. In permeabilized cells, the ODN readily migrated into the nucleus where, at ambient temperature, preferentially the S-ODN gave rise to a multitude of large, irregular aggregates. Nuclear uptake of the ODN was considerably and differentially inhibited by wheat germ agglutinin. High-affinity S-ODN, but not P-ODN, additionally reacted with a structure presumably identical with the nuclear lamina. Simultaneously, they cause decompaction of chromatin, whereby the S-ODN aggregates appeared as compact inclusions in homogeneously dispersed chromatin. After microinjection of S-ODN into intact cells, these effects were not observed, although the nucleic acids rapidly moved into the nucleus and condensed into a large number of well-defined, spherical speckles or longitudinal rodlets. The methylphosphonate analogs of some of the ODN used exhibited only extremely low affinities for intracellular constituents. These results show that excess amounts of S-ODN saturate a host of both low-affinity and high-affinity binding sites on cellular substructures, whereas limited quantities as used for microinjection recognize only the high-affinity binding sites. The results support the notion that the nonsequence-specific, often toxic effects of antisense S-ODN result from their strong binding to cellular components and substructures involved in replicational, transcriptional, and translational processes. On the other hand, the association of the ODN with membranes and cytoskeletal and karyoskeletal elements may serve to optimize their sequence-specific interaction with their intended target sites and also increase their cellular retention potential. These cellular structures would thus fulfill a depot function.

Actins↗
PubMed2003

Targeted deletion of the lens fiber cell-specific intermediate filament protein filensin.

PURPOSE: To determine the function of the lens fiber cell-specific cytoskeletal protein, filensin, in lens biology. METHODS: Targeted genomic deletion was used to delete exon 1 and the transcriptional start site of the filensin gene. Resultant chimeric animals were bred to homozygosity for the mutant allele. These animals were outbred to mice bearing the wild-type CP49 alleles to eliminate the mutant CP49 gene carried by the 129 strain of mice. Animals homozygous for the mutated filensin gene and wild-type CP49 gene were compared with wild-type and heterozygous animals by Northern and Western blot analyses, light and electron microscopy, and slit lamp microscopy. RESULTS: Disruption of the filensin gene successfully blocked production of filensin mRNA, reduced levels of filensin's assembly partner CP49, and prevented the assembly of beaded filaments. Despite the absence of beaded filaments, lenses did not show obvious changes in fetal development, nor in the differentiation of epithelial cells into mature fiber cells, as judged by light microscopic analysis. Filensin knockouts began to show evidence of light-scattering by 2 months and worsened with age. Heterozygous animals exhibited an intermediate phenotype, showing a reduction in filensin transcript and moderate light-scattering at 5 months. CONCLUSIONS: The lens fiber cell-specific intermediate filament protein filensin is essential for beaded filament assembly. However, although beaded filaments are not needed for normal lens fetal development or fiber cell differentiation, they appear to be necessary for the long-term maintenance of optical clarity. The mechanism by which the absence of filensin and the beaded filament affects optical clarity has yet to be defined.

Animals↗
PubMedDate not supplied

Homologies in both primary and secondary structure between nuclear envelope and intermediate filament proteins.

The A, B and C lamins are the major proteins of the nuclear envelope. The complete nucleotide sequence of the coding region of the A and C lamins shows that these proteins are identical except for their carboxy termini. The most prominent structural feature of both lamins is an alpha-helical region of repeating heptads of amino acids that shows striking homology with the entire family of cytoplasmic intermediate filament proteins. These features suggest that the nuclear envelope is made up of a network of coiled-coil polymers.

Amino Acid Sequence↗
PubMed1985

Characterization of rat mammary cell types in primary culture: lectins and antisera to basement membrane and intermediate filament proteins as indicators of cellular heterogeneity.

Three morphologically distinct major cell types were observed in primary cultures obtained from the mammary parenchyma of glands from virgin rats. These cell types consisted of small cuboidal epithelial cells, larger epithelioid cells and elongated cells. We have investigated the distribution of the basement membrane proteins laminin and type IV collagen, and the intermediate filament proteins vimentin and prekeratin, in these three cell types using immunofluorescence techniques. Antisera to the basement membrane proteins stain the large epithelioid cells and the elongated cells, but do not stain the small cuboidal cells. Polyclonal antiserum to keratin stains all the small cuboidal and large epithelioid cells, but only a small subpopulation of the elongated cells. However, a monoclonal antibody to keratin, LP34, stains only the large cuboidal and a proportion of the elongated cells. Vimentin antiserum fails to stain the small cuboidal cells but stains all the large epithelioid and elongated cells. In addition, peanut lectin, which binds only to ductal lining epithelial cells in the virgin rat mammary gland in vivo after their treatment with neuraminidase, binds to the small cuboidal cells after neuraminidase treatment but not to the other cell types. However, Griffonia simplicifolia agglutinin I, which specifically stains myoepithelial cells in vivo, binds to the large epithelioid and elongated cells but not to the small cuboidal cells. These results suggest that the small cuboidal cells are related to mammary ductal epithelial cells whereas the large epithelial and elongated cells have some characteristics of myoepithelial cells.

Animals↗
PubMed1987

Platelet intermediate filaments: detection of a vimentinlike protein in human and bovine platelets.

Human and bovine platelets contain a 58,000-dalton vimentinlike protein that cross-reacts with antivimentin antibody. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blots indicate that this protein is present in whole platelet lysates and triton insoluble cytoskeletons. Transmission electron microscopy of platelets reveals an isotropic network of individual intermediate filaments distributed throughout the platelets. High salt, triton extracted, glutaraldehyde and tannic acid fixed platelets reveal 10-nm filaments that can be seen to form a peripheral ring, as well as an isotropic network in the body of the cells. Indirect immunofluorescence of resting and spread platelets demonstrates a circumferential staining pattern close to the cell membrane, with additional fibrillar staining throughout the platelets. Our data suggest that the 58,000-dalton vimentinlike protein may be associated with the microtubule coil and the plasma membrane, and may thus help to maintain the resting platelet's discoid shape.

Animals↗
PubMed1988

A nerve growth factor-regulated messenger RNA encodes a new intermediate filament protein.

Differential screening of a cDNA library from the PC12 rat pheochromocytoma cell line previously revealed a clone, clone 73, whose corresponding mRNA is induced by nerve growth factor (NGF). Induction parallels NGF-stimulated PC12 differentiation from a chromaffinlike phenotype to a sympathetic neuronlike phenotype. We report that DNA sequence analysis reveals that clone 73 mRNA encodes an intermediate filament (IF) protein whose predicted amino acid sequence is distinct from the known sequences of other members of the IF protein family. The sequence has highest homology with desmin and vimentin and includes the highly conserved central alpha-helical rod domain with the characteristic heptad repeat of hydrophobic residues, but has lower homology in the amino-terminal head and carboxyl-terminal tail domains. The head domain contains a large number of serine residues which are potential phosphorylation sites. The expression of clone 73 in vivo in the nervous system of the adult rat was investigated by in situ hybridization of clone 73 probes to tissue sections. The mRNA is expressed at high levels in ganglia of the peripheral nervous system, including the superior cervical ganglion (sympathetic), ciliary ganglion (parasympathetic), and dorsal root ganglion (sensory). In the central nervous system, motor nuclei of cranial nerves III, IV, V, VI, VII, X, and XII as well as ventral horn motor neurons and a restricted set of other central nervous system nuclei express the clone 73 mRNA. Tissues apart from those of the nervous system did not in general express the mRNA, with only very low levels detected in adrenal gland. We discuss the implications of these results for the mechanism of NGF-induced PC12 cell differentiation, the pathways of neuronal development in vivo, and the possible function of the clone 73 IF protein and its relationship to other IF proteins.

Amino Acid Sequence↗
PubMedDate not supplied

Proteins of the intermediate filament cytoskeleton as markers for astrocytes and human astrocytomas.

There is a pressing need for a more accurate system of classifying human astrocytomas, one that is based on morphologic characteristics and that could also make use of distinctive biochemical markers. However, little is known about the phenotypic characteristics of astrocytomas. Recent studies have shown that the expression of proteins comprising the intermediate filament (IF) cytoskeleton of astrocytic cells is developmentally regulated. It is our hypothesis that this changing protein profile can be used as the basis of a system for clearly and objectively classifying astrocytomas. A spectrum of human astrocytomas has been examined by immunofluorescence microscopy employing antibodies to several IF structural subunit proteins (GFAP, vimentin, and keratins) and an IF-associated protein, IFAP-300kDa. These proteins occupy unique temporal niches in the cytogenesis of the astrocytic cells: keratins in cells of the neuroectoderm; vimentin and IFAP-300kDa in radial glia and immature glia; GFAP in mature astrocytes; and vimentin in some mature astrocytes. In agreement with previous reports, our immunofluorescence studies have revealed both GFAP and vimentin in all astrocytoma specimens. Two new observations, however, are of particular interest: IFAP-300kDa is detectable in all astrocytic tumors, and the proportion of keratin-containing cells present in the astrocytomas is in direct relationship to the degree of the malignancy. Because IFAP-300kDa is not present in either normal mature or reactive astrocytes, this protein appears to represent a specific marker of transformed (malignant) astrocytes. If it is presumed that higher malignancy grades represent the most dedifferentiated cellular state of the astrocytes, the presence of keratin-containing cells is not totally unexpected, given the ectodermal (epithelial) origin of the CNS. Specific developmentally regulated proteins of the IF cytoskeleton thus appear to hold great potential as diagnostic markers of astrocytomas and as tools for investigating the biology of these tumors.

Astrocytes↗
PubMed1986

Characterization of the intermediate filament proteins of murine mammary gland epithelial cells. Response to collagen substratum.

The insoluble cytoskeletal material remaining after detergent lysis of 'Normal' Murine Mammary Gland (NMuMG) cells, growing on plastic or collagen gel substrata, was analyzed by two-dimensional gel electrophoresis. The identity of the cytoskeletal elements was determined by their solubility properties, electrophoretic separation pattern, and immunoreactivity using monoclonal antibodies against intermediate filament proteins (AIF), keratins (AE1 and AE3) and actin. The electrophoretic pattern of the cytoskeletal elements from the NMuMG cell strain was found to be very similar to that of primary mouse mammary epithelial cells. Both NMuMG and primary mammary epithelial cells when grown on collagen exhibited an increased expression of a 49 kD protein with a pI of 5.6, that appeared to be a cytokeratin. Many of the cytoskeletal proteins remained tightly attached to the collagen gel substratum after cell lysis. These results demonstrate that the NMuMG cell strain has retained a stable expression of cytokeratins that remains responsive to the presence of extracellular matrix material.

Animals↗
PubMed1986

Activation of complement by intermediate filaments of glomerular epithelial cells.

Glomerular visceral epithelial cells, podocytes, have been shown by immunofluorescence and immunoperoxidase staining methods to bind serum complement (C) components in vitro. Binding of Clq, C4, and C3 was demonstrated and the C4 and C3 binding could be inhibited by EDTA. Thus complement binding had the capacity to cause antibody-independent activation of the "classical" C pathway. The C-binding structures had the same distribution as intermediate filaments (IMFs) of vimentin but not desmin or keratin types. By ultrastructural immunohistochemical staining C3 was observed in association with cytoskeletal IMFs.

Complement Activation↗