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E Rungger-Brändle

Publications and source records attributed to E Rungger-Brändle.

35 records · Page 2Linked to original sources

Association of specific cell-surface glycoproteins with a triton X-100-resistant complex of plasma membrane proteins isolated from T-lymphoma cells (P1798).

A non-ionic detergent-resistant complex of membrane-associated proteins and cell-surface glycoproteins has been isolated by gel filtration and isopyknic centrifugation of purified plasma membranes from the murine T-lymphoma P 1798. This complex elutes as a high molecular weight peak (greater than 15 X 10(6) D) and contains two specific sets of (1) cell-surface glycoproteins; (2) membrane-associated proteins. The cell-surface glycoproteins consist of two vectorially labelled major components present in a fixed molar ratio: The Thy-1 glycoprotein and a non-H-2 glycoprotein of 55 kD. Minor but significant amounts of the class I histocompatibility antigen Qa-2 are also contained in the detergent-resistant complex. The membrane-associated proteins are not vectorially labelled, and form a complex group of proteins in the 30-70 kD range. Since actin is not detectable among these polypeptides, they probably constitute a plasma membrane-associated structure that is distinct from actin-containing, submembranous cytoskeletal elements.

Animals↗

Bilateral acute retinal necrosis (BARN). Identification of the presumed infectious agent.

We describe histopathologic features of an enucleated eye of a patient suffering bilateral acute retinal necrosis (BARN). Retinal tissue was found focally degenerated, and the choroid massively enlarged by lymphoid-like agranular cells. An association of the disease with a viral infection could be demonstrated by (a) the presence of virus particles of the herpesvirus type in retinal tissue, (b) the transmission of the infected principle to human embryo fibroblast cultures, and (c) the visualization of CMV-antigens by immunofluorescence microscopy in such infected cultures. Slow growth of the virus in vitro and the presence of CMV-antigens after infection indicate that the herpesvirus involved in BARN was of the type CMV. On the basis of these findings we propose a guideline for therapy.

Acute Disease↗

Isolation of plasma membrane domains from murine T lymphocytes.

Murine T-lymphoma cells have been homogenized in dense sucrose solution and centrifuged under isopycnic conditions for membrane components. Floating fractions banding between 10% and 22.5% sucrose ("light" membranes) and between 22.5% and 35% sucrose ("heavy" membranes) were shown to consist of smooth membrane vesicles. The amounts of cholesterol and phospholipids recovered after chloroform/methanol extraction were similar in both fractions, but heavy membranes contained two to three times more protein than light membranes. The most striking difference between the two membrane fractions was revealed by their labeled surface glycoprotein patterns on polyacrylamide gels, suggesting that (i) the smooth membrane vesicles originated from the plasma membrane and (ii) two distinct segments of the plasma membrane can be recovered in fractions characterized by specific surface glycoproteins. Light membranes were enriched in Thy-1 antigen, whereas Ly-5 antigen and a 170,000-dalton surface glycoprotein were recovered almost exclusively from heavy membranes, as were metabolically labeled protein spots comigrating with the H-2k/d antigen in two-dimensional electrophoresis. The patterns of the unlabeled proteins in light and heavy membranes appeared similar, except for polypeptides of 180,000 and 85,000 daltons that were found preferentially in heavy membranes. These results support the concept of plasma membrane domains by showing that two distinct populations of plasma membrane vesicles can be isolated and that these populations contain different sets of cell surface glycoproteins.

Animals↗

Proteins of intermediate filaments. An immunohistochemical and biochemical approach to the classification of soft tissue tumors.

The intermediate filament cytoskeleton of various types of human soft tissue tumors was analyzed by immunofluorescence microscopy with the use of specific antibodies against cytokeratins, vimentin, and desmin, as well as by one- and two-dimensional gel electrophoresis of high-salt buffer- and detergent-resistant cytoskeletal preparations. All leiomyomas as well as a leiomyosarcoma contained desmin. Leiomyomas of both gastrointestinal and uterine derivation and the retroperitoneal leiomyosarcoma showed strong reaction for desmin in the smooth muscle cells, but the latter two exhibited also vimentin staining. In embryonal rhabdomyosarcomas, desmin prevailed in the large, apparently well-differentiated rhabdomyoblasts; whereas the smaller, less differentiated tumor cells preferentially contained vimentin. Cells of malignant fibrous histiocytomas were characterized by their content of vimentin as the only intermediate filament protein present. In alveolar soft part sarcoma, a rare tumor of hitherto unknown histogenesis, vimentin and desmin co-existed within the same tumor cells, indicating, together with chemical determinations, the myogenic derivation of this neoplasm. The results show that immunologic and biochemical analysis of proteins associated with the intermediate filament cytoskeleton is a useful adjunct in the diagnosis of diverse neoplasms, particularly those with equivocal histologic features, and thus aids in the histogenetic classification of soft tissue tumors.

Desmin↗

The area of attachment of cytotoxic T lymphocytes to their target cells shows high motility and polarization of actin, but not myosin.

Conjugates of cytotoxic T lymphocytes attached to their target cells were studied by double immunofluorescence on fixed smears to detect simultaneously the localization of actin and myosin within the cells. Actin was found to be polarized in the area of attachment of the lymphocytes (but not of the target cells), whereas myosin remained evenly distributed in the cytoplasm. When living conjugates were brought to 37 degrees C to induce cytotoxicity, this pattern remained unchanged, but observation by interference reflexion microcinematography revealed a high motility of the lymphocytes in the contact area. This localized motility in the area of attachment associated with a peculiar actin polarization, which has no equivalent in any type of cell contact presently known, could represent a necessary step in the sequence of events leading to target cell killing by cytotoxic T lymphocytes.

Actins↗

Vimentin-containing smooth muscle cells in aortic intimal thickening after endothelial injury.

We evaluated by means of (1) indirect immunofluorescence and (2) densitometric analysis of stained sodium dodecyl sulfate-polyacrylamide gels the distribution and amount of vimentin and desmin in the aortic media and intima of normal rats and of rats in which the endothelium had been removed by means of an inflated balloon catheter. According to previous reports, in normal aorta vimentin was found in endothelial cells and in the majority of smooth muscle cells, and desmin was found in several smooth muscle cells. Smooth muscle cells present in areas of intimal thickening in rats whose aortas were injured with a balloon were stained only with vimentin antibodies, whereas in the media underlying intimal thickening, the distribution of vimentin and desmin was similar to that seen in normal rats. Portions of aorta (media plus intima) with intimal thickening had a significantly increased vimentin to desmin ratio compared with similar portions of aorta in normal rats. These results suggest that after endothelial injury either only vimentin-containing smooth muscle cells are responsible for replication and migration into the intima, or that smooth muscle cells, while migrating into the intima, synthesize only vimentin.

Animals↗

Localization of RNA polymerase B and histones in the nucleus of primary spermatocytes of Drosophila hydei, studied by immunofluorescence microscopy.

By means of indirect immunofluorescence microscopy, we have studied the distribution of RNA polymerase B, of the nucleosomal histones H2b, H3, and H4 and of histone H1, in nuclei of primary spermatocytes of Drosophila hydei. RNA polymerase B and histones, including H1, are found to be present on the loop structures of the Y chromosome. The nucleolus stains only for the histones, but not for RNA polymerase B. Various mutants deficient for some of the loops or altering their morphology, were used to identify the individual chromosomal segments. In growing spermatocytes of the genetic constitution X/0, autosomes and the chromosome X react strongly with antibodies against RNA polymerase B, but not with antibodies against histones. The results suggest that the autosomes, the chromosome X and the Y chromosomal loop structures, with the exception of the nucleolus, are transcribed mostly by RNA polymerase B.

Animals↗

Distribution of cytoskeletal elements in cultured skin fibroblasts of patients with Duchenne's muscular dystrophy.

Cultured fibroblasts from patients suffering from Duchenne's Muscular Dystrophy were examined by indirect immunofluorescent techniques using antibodies against actin, myosin, tubulin, and intermediate-sized filaments. The cells display normal patterns of microfilamentous bundles (stress fibres), microtubules, and intermediate-sized filaments suggesting a normal organization of these cytoskeletal structures.

Actins↗

Lymphocyte alpha-actinin. Relationship to cell membrane and co-capping with surface receptors.

Mouse spleen lymphocytes synthesize a protein which comigrates with skeletal muscle alpha-actinin on two-dimensional gel electrophoresis and is immunoprecipitated by an antibody directed against skeletal muscle alpha-actinin. Mouse lymphocyte alpha-actinin is present in membrane fractions, and is immunoprecipitated from lymphocyte detergent lysates by an antiserum made against these purified membranes. The anti-alpha-actinin activity of this antiserum is not adsorbed after incubation with fixed intact lymphocytes. Lymphocyte alpha-actinin does not bind concanavalin A and it is inaccessible to lactoperoxidase-catalyzed surface iodination. Double immunofluorescence shows that alpha-actinin moves concurrently along the cell membrane with redistributed surface immunoglobulins and Thy-1 antigen, and remains associated up to 30 min with surface aggregates of these receptors. Our results suggest that lymphocyte alpha-actinin, as defined by molecular weight and cross reactivity with the antibody against the muscle protein, (a) is associated with the cell membrane, (b) is not expressed at the cell surface, and (c) participates in the movement of surface receptors.

Actinin↗

Intranuclear injection of anti-actin antibodies into Xenopus oocytes blocks chromosome condensation.

The role of contractile proteins in the structural organisation of the interphase nucleus and of metaphase chromosomes is largely unknown. Actin has been found in interphase nuclei of different species, especially in association with condensed chromatin. In the germinal vesicle (nucleus) of Xenopus oocytes, actin has been localised in the nuclear gel supporting the chromosomes and the extrachromosomal nucleoli. It has been reported that the premeiotic lampbrush chromosomes in these germinal vesicles are positively stained for actin and tubulin by the immunoperoxidase technique. Moreover, the longitudinal contraction of these chromosomes is ATP dependent. Therefore it has been suggested that actin participates in the structural organisation of the highly specialised lampbrush chromosomes. However, actin is not a major component of the metaphase chromosome scaffold. The results reported here suggest that actin is involved in the condensation of Xenopus chromosomes.

Actins↗

An actin-destabilizing factor is present in human plasma.

Plasma and serum of humans or experimental animals contain a factor which destabilizes F-actin. The factor has no DNAse or thrombin activity and after incubation with F-actin does not modify the position of the actin band on a SDS polyacrylamide gel. Hence it probably depolymerizes F-actin.

Actins↗

Indirect immunofluorescence microscopy of microtubular structures in male germ cells of wildtype and l(3)pl (lethal-polyploid) Drosophila hydei.

Tubulin-containing structures of the male germ cells of Drosophila hydei crossreact in indirect immunofluorescence microscopy with antibody directed against homogeneous porcine brain tubulin. There is no detectable difference in reactivity between germ cells of wildtype flies and the mutant l(3)pl (lethal-polyploid) which is characterized by microtubular abnormalities. However, the technique of indirect immunofluorescence microscopy allows the direct visualization of several abnormalities in the arrangement of the microtubular system of the mutant, particularly in the axonemal complex.

Animals↗