Your email was sent successfully. Check your inbox.

An error occurred while sending the email. Please try again.

Proceed reservation?

Export
Filter
  • AREA-COMPOSITA  (5)
  • Germany  (4)
  • MOUSE MODEL  (3)
  • 1
    Keywords: CELLS ; GENE-TRANSFER ; MOUSE MODEL ; DEFICIENT MICE ; ENZYME REPLACEMENT THERAPY ; ALPHA-GALACTOSIDASE-A ; SUBSTRATE REDUCTION THERAPY ; TYPE-1 GAUCHER-DISEASE ; GLOBOTRIAOSYLCERAMIDE ; ISOGLOBOTRIHEXOSYLCERAMIDE
    Abstract: Fabry disease is a monogenic X-linked lysosomal storage disease caused by alpha-galactosidase A (alphaGalA) deficiency. Enzyme replacement therapy through administration of the missing alphaGalA is currently the only accepted therapeutic option. However, this treatment is connected to high costs, has ill-defined indication criteria and its efficacy is controversially discussed. Our aim was to explore the possibility of a novel targeted substrate reduction therapy for Fabry disease. Owing to the fact that alphaGalA-deficient humans and mice accumulate the same glycosphingolipids (i.e. globosides, galabiosylceramide and isoglobosides), alphaGalA-deficient mice were crossed with mice deficient in enzymes synthesizing these classes of glycosphingolipids (i.e. globotrihexosylceramide and isoglobotrihexosylceramide synthase, respectively). Functional heart and kidney tests were performed together with an extensive biochemical analysis of urine and serum in aged mice. Lysosomal storage was assessed by thin layer chromatography and electron microscopy. We showed that depletion of globosides was sufficient to fully abolish the storage of glycosphingolipids in heart, kidney and liver and was paralleled by a complete restoration of lysosomal morphology in these organs. In contrast, in dorsal root ganglia, a depletion of both globosides and isoglobosides was necessary to fully counteract the lysosomal storage. The deficiency in globosides and/or isoglobosides did not cause any adverse effects. We conclude that substrate reduction therapy through inhibition of the synthesis of globosides and isoglobosides represents a valuable therapeutic option for Fabry disease, all the more as globosides and isoglobosides seem to be dispensable.
    Type of Publication: Journal article published
    PubMed ID: 24992926
    Signatur Availability
    BibTip Others were also interested in ...
  • 2
    Keywords: RIGHT-VENTRICULAR CARDIOMYOPATHY ; DESMOPLAKIN-CONTAINING JUNCTIONS ; adhering junctions ; INTERCALATED DISC ; HEART-MUSCLE CELLS ; AREA-COMPOSITA ; PROTEIN PHOSPHATASE 2A ; ARRHYTHMOGENIC CARDIOMYOPATHY ; CALMODULIN-BINDING PROTEIN ; SODIUM CURRENT DEFICIT
    Abstract: Proteins of the striatin family (striatins 1-4; sizes ranging from 90 to 110 kDa on SDS-polyacrylamide gel electrophoresis) are highly homologous in their amino acid sequences but can differ in their cell-type-specific gene expression patterns and biological functions. In various cell types, we have found one, two or three polypeptides of this evolutionarily old and nearly ubiquitous family of proteins known to serve as scaffold proteins for diverse protein complexes. Light and electron microscopic immunolocalization methods have revealed striatins in mammalian cell-cell adherens junctions (AJs). In simple epithelia, we have localized striatins as constitutive components of the plaques of the subapical zonulae adhaerentes of cells, including intestinal, glandular, ductal and urothelial cells and hepatocytes. Striatins colocalize with E-cadherin or E-N-cadherin heterodimers and with the plaque proteins alpha- and beta-catenin, p120 and p0071. In some epithelia and carcinomas and in cultured cells derived therefrom, striatins are also seen in lateral AJs. In stratified epithelia and in corresponding squamous cell carcinomas, striatins can be found in plaques of some forms of tessellate junctions. Moreover, striatins are major plaque proteins of composite junctions (CJs; areae compositae) in the intercalated disks connecting cardiomyocytes, colocalizing with other CJ molecules, including plectin and ankyrin-G. We discuss the "multimodulator" scaffold roles of striatins in the initiation and regulation of the formation of various complex particles and structures. We propose that striatins are included in the diagnostic candidate list of proteins that, in the CJs of human hearts, can occur in mutated forms in the pathogeneses of hereditary cardiomyopathies, as seen in some types of genetically determined heart damage in boxer dogs.
    Type of Publication: Journal article published
    PubMed ID: 25501894
    Signatur Availability
    BibTip Others were also interested in ...
  • 3
    Keywords: brain ; CELLS ; ENDOTHELIAL-CELLS ; EXPRESSION ; IN-VITRO ; CELL ; Germany ; human ; MODEL ; VITRO ; GENE ; GENES ; PROTEIN ; cell line ; TISSUE ; MARKER ; BIOLOGY ; E7 ; immunohistochemistry ; resistance ; CELL-LINE ; LINE ; human papillomavirus ; E6 ; HUMAN-PAPILLOMAVIRUS ; MORPHOLOGY ; BARRIER FUNCTION ; TIGHT JUNCTIONS ; CONJUGATE EXPORT PUMP ; MRP2 ; ORIGIN ; RE ; TRANSFECTION ; P-GLYCOPROTEIN ; blood-brain barrier ; human brain ; ENDOTHELIAL-CELL ; brain capillary endothelial cell ; ATP-BINDING ; BLOOD-BRAIN ; ABC-TRANSPORTERS ; DRUG EFFLUX TRANSPORTERS ; endothelial markers
    Abstract: Primary human brain capillary endothelial cells (hBCECs) are available only in small quantities and have a short life span in vitro; this restricts their use as in vitro model for the blood-brain barrier (BBB). To overcome these limitations, we have established an immortalized hBCEC line (NKIM-6) by transfection with pLXSN16-E6E7, which encodes the human papillomavirus type 16 E6 and E7 genes. The cell line exhibits an extended life span in vitro and retains its characteristic endothelial morphology, endothelial markers, and physiology. Likewise, as demonstrated by immunohistochemistry and reverse transcription/polymerase chain reaction (RT-PCR), NKIM-6 cells express BBB markers, and the lack of glial, neuronal, and epithelial markers confirms their endothelial origin. Moreover, with quantitative RT-PCR, we have been able to demonstrate that several ATP-binding cassette-transporters are expressed in NKIM-6 cells with a conserved expression order compared with primary hBCECs. Our results suggest that this cell line might be suitable as in vitro model for several aspects of the BBB
    Type of Publication: Journal article published
    PubMed ID: 17180596
    Signatur Availability
    BibTip Others were also interested in ...
  • 4
    Keywords: brain ; CELLS ; EXPRESSION ; IN-VITRO ; SURVIVAL ; AGENTS ; Germany ; MODEL ; THERAPY ; GENE ; PROTEIN ; PROTEINS ; gene therapy ; MICE ; TUMOR-NECROSIS-FACTOR ; IFN-GAMMA ; INFECTION ; ALPHA ; culture ; MOUSE ; VECTOR ; NECROSIS-FACTOR-ALPHA ; CELL-LINE ; LINE ; MINUTE VIRUS ; REPLICATION ; MOUSE MODEL ; INTERFERON ; sensitivity ; autonomous parvovirus ; mannose receptor ; CYTOTOXICITY ; FACTOR-ALPHA ; BRAIN-TUMORS ; CAPACITY ; GLIOMA ; parvovirus ; VIRIONS ; FUNCTIONAL-CHARACTERIZATION ; astrocytoma ; uptake ; CANDIDATE ; glial cells ; IMMUNE FUNCTION ; lipopolysaccharide ; MICROGLIAL CELLS
    Abstract: The sensitivity of brain tumour cells to wild-type or recombinant parvoviruses H1-PV and MVMp makes these agents promising candidates for gene therapy of astrocytoma. This application raises the question of whether parvoviruses exert deleterious or bystander effects on normal glial cells surrounding tumours. We addressed this question in the mouse model by using cell cultures derived from BALB/c, C57BL/6 and VM/Dk strains. Astrocytes and a large proportion of microglia cultures were competent for MVMp uptake. Infection was, however, abortive as replication-associated viral proteins synthesis took place in less than 10% of astrocytes and no progeny virions were produced. This restriction was even more pronounced for microglia in which no viral protein expression could be detected, save for a minute fraction of VM/Dk-derived cells. Infection with MVMp had no significant effect on glial cell survival and did not interfere with their immune potential. Indeed, neither the lipopolysaccharide (LPS)/interferon (IFN-gamma)-induced cytotoxicity of VM/Dk-derived microglia towards the mouse glioma (MT539MG) cell line, nor the glial cells capacity for tumour necrosis factor alpha production upon LPS stimulation or LPS/IFN-gamma stimulation were affected by infection with MVMp. Moreover, stimulation with LPS and/or IFN-gamma resulted in a decreased expression of the viral replicative and cytotoxic protein NS1. Together, our data indicate that, in the natural host, a majority of normal glial cells are not competent for MVMp replication and that the abortive infection taking place in a minor fraction of these cells fails to impede their survival and immunocompetence, giving credit to the consideration of autonomous parvoviruses for glioma therapy
    Type of Publication: Journal article published
    PubMed ID: 16699801
    Signatur Availability
    BibTip Others were also interested in ...
  • 5
    Keywords: immunohistochemistry ; ADHESION ; TIGHT JUNCTIONS ; HUMAN EPIDERMIS ; desmosomes ; STRATIFIED EPITHELIA ; ADHERENS JUNCTIONS ; cardiomyocytes ; HEART-MUSCLE CELLS ; plakophilin-2 ; AREA-COMPOSITA ; VERTEBRATES ; Epithelial cells ; Junction plaques ; Protein myozap
    Abstract: The protein myozap, a polypeptide of 54 kDa, has recently been identified as a component of the cytoplasmic plaques of the composite junctions (areae compositae) in the myocardiac intercalated disks and of the adherens junctions (AJs) in vascular endothelia. Now we report that using very sensitive new antibodies and drastic localization methods, we have also identified this protein as a component of the AJ plaques in simple and complex epithelia, in the adluminal cell layer of the transitional epithelium of the urinary tract and in certain cell layers of diverse stratified epithelia, including gingiva, tongue, pharynx and esophagus, cervix, vagina and epidermis. Myozap has not been identified in desmosomal and tight junction plaques. We have also detected protein myozap in AJ structures of carcinomas. The discovery of a novel major protein in AJ plaques now calls for re-examinations of molecular interactions in AJ formation and maintenance and also offers a new marker for diagnostic immunocytochemistry. We also discuss the need for progressive unravelling, extractive treatments and buffer rinses of sections and cultured cells to reveal obscured or masked antigens, before definitive negative conclusions in immunohistochemistry can be made.
    Type of Publication: Journal article published
    PubMed ID: 22160502
    Signatur Availability
    BibTip Others were also interested in ...
  • 6
    Keywords: TISSUE ; TUMORS ; EXTRACELLULAR-MATRIX ; RIGHT-VENTRICULAR CARDIOMYOPATHY ; ADHERENS JUNCTIONS ; adhering junctions ; MESENCHYMAL STEM-CELLS ; plakophilin-2 ; AREA-COMPOSITA ; Puncta adhaerentia ; RECESSIVE MUTATION ; Heart valves ; MUSCLE CELLS ; PORCINE AORTIC-VALVE ; ATRIOVENTRICULAR VALVES ; CARDIAC-VALVE ; CUSHION DEVELOPMENT ; Myxomata ; Valvular interstitial cells
    Abstract: The interstitial cells of cardiac valves represent one of the most frequent cell types in the mammalian heart. In order to provide a cell and molecular biological basis for the growth of isolated valvular interstitial cells (VICs) in cell culture and for the use in re-implantation surgery we have examined VICs in situ and in culture, in fetal, postnatal and adult hearts, in re-associations with scaffolds of extracellular matrix (ECM) material and decellularized heart valves. In all four mammalian species examined (human, bovine, porcine and ovine), the typical mesenchymal-type cell-cell adherens junctions (AJs) connecting VICs appear as normal N-cadherin based puncta adhaerentia. Their molecular ensemble, however, changes under various growth conditions insofar as plakophilin-2 (Pkp2), known as a major cytoplasmic plaque component of epithelial desmosomes, is recruited to and integrated in the plaques of VIC-AJs as a major component under growth conditions characterized by enhanced proliferation, i.e., in fetal heart valves and in cell cultures. Upon re-seeding onto decellularized heart valves or in stages of growth in association with artificial scaffolds, Pkp2 is - for the most part - lost from the AJs. As Pkp2 has recently also been detected in AJs of cardiac myxomata and diverse other mesenchymal tumors, the demonstrated return to the normal Pkp2-negative state upon re-association with ECM scaffolds and decellularized heart valves may now provide a safe basis for the use of cultured VICs in valve replacement surgery. Even more surprising, this type of transient acquisition of Pkp2 has also been observed in distinct groups of endothelial cells of the endocardium, where it seems to correspond to the cell type ready for endothelial-mesenchymal transition (EMT)
    Type of Publication: Journal article published
    PubMed ID: 22290634
    Signatur Availability
    BibTip Others were also interested in ...
  • 7
    Keywords: PEPTIDE ; CELLS ; EXPRESSION ; Germany ; human ; KINASE ; MICROSCOPY ; NEW-YORK ; PROTEIN ; PROTEINS ; cell line ; TISSUE ; FAMILY ; ACTIVATED PROTEIN-KINASE ; PHOSPHORYLATION ; treatment ; cell culture ; ACIDS ; antibodies ; antibody ; MAP KINASE ; DISRUPTION ; PLASMA ; MEMBRANE ; STRESS ; LOCALIZATION ; CHROMATOGRAPHY ; AMINO-ACIDS ; PHORBOL-ESTER ; serine ; CHANNEL ; GAP-JUNCTIONAL COMMUNICATION ; growth factors ; hyperosmosis ; INTERCELLULAR COMMUNICATION ; LIVER EPITHELIAL-CELLS ; plasma membranes ; sorbitol ; threonine phosphorylation
    Abstract: We have developed polyclonal antibodies (SA226P) to a peptide of the human connexin43 (Cx43) protein between amino acids 271 and 288 containing phosphorylated S279 and S282. Antibodies specific for the phosphorylated form of the peptide were isolated by double immunoaffinity chromatography and were characterised using proteins of the cell line WB-F344, known to contain large amounts of Cx43. SA226P recognises specifically the slowest migrating Cx43 band in immunoblots of proteins isolated from untreated cells. In immunofluorescence experiments SA226P scarcely stains the plasma membrane in untreated cells in contrast to a commercial antibody recognising all isoforms of the Cx43 protein. EGF or stress treatment of the cells results in a rapid increase in the phosphorylated forms of Cx43 as revealed by immunoblotting. Immunofluorescence experiments reveal that both phosphorylated and non-phosphorylated Cx43 could be found at the plasma membrane. Whether phosphorylation of S279/S282 takes place before or after incorporation of Cx43 into the membranes is so far unknown. More interestingly, confocal microscopy using our antibodies and a commercial antibody recognising all isoforms of Cx43 shows the coexistence of differentially phosphorylated forms of the protein at the plasma membrane. Our results indicate that MAP kinases erk1/2 are mainly responsible for this phosphorylation, as already published. Nevertheless, treatment of the cells with anisomycin, known to activate stress kinase p38 but not erk1/2, also results in a weak but reproducible Cx43 phosphorylation
    Type of Publication: Journal article published
    PubMed ID: 12483281
    Signatur Availability
    BibTip Others were also interested in ...
  • 8
    Keywords: ANGIOGENESIS ; CELLS ; ENDOTHELIAL-CELLS ; IN-VITRO ; BLOOD ; Germany ; human ; IN-VIVO ; VITRO ; SYSTEM ; PROTEIN ; PROTEINS ; desmoplakin ; TISSUE ; COMPLEX ; COMPLEXES ; TISSUES ; beta-catenin ; TIGHT JUNCTIONS ; rodent ; JUNCTIONS ; LYMPHATIC ENDOTHELIUM ; RE ; DESMOSOMAL PLAQUE ; INTERCELLULAR-JUNCTIONS ; DESMOSOMAL PLAQUE PROTEINS ; SIZE ; ADHERENS JUNCTIONS ; BOVINE ; function ; lymph node ; LYMPH-NODE ; N-CADHERIN ; adhering junction ; VE-CADHERIN ; BLOOD-BRAIN-BARRIER ; Complexus adhaerens ; DESMOPLAKIN-CONTAINING JUNCTIONS ; HUMAN GLIOBLASTOMA-MULTIFORME ; retothelium ; TO-CELL JUNCTIONS
    Abstract: The significance of a special kind of VE-cadherin-based, desmoplakin- and plakoglobin-containing adhering junction, originally identified in certain endothelial cells of the mammalian lymphatic system ( notably the retothelial cells of the lymph node sinus and a subtype of lining endothelial cells of peripheral lymphatic vessels), has been widely confirmed and its importance in the formation of blood and lymph vessels has been demonstrated in vivo and in vitro. We have recently extended the molecular and structural characterization of the complexus adhaerens and can now report that it represents a rare and special combination of components known from three other major types of cell junction. It comprises zonula adhaerens proteins (VE-cadherin, alpha- and beta-catenin, protein p120(ctn), and afadin), desmosomal plaque components ( desmoplakin and plakoglobin), and tight-junction proteins (claudin-5 and ZO-1) and forms junctions that vary markedly in size and shape. The special character and the possible biological roles of the complexus adhaerens and its unique ensemble of molecules in angiogenesis, immunology, and oncology are discussed. The surprising finding of claudin-5 and protein ZO-1 in substructures of retothelial cell-cell bridges, i.e., structures that do not separate different tissues or cell layer compartments,suggests that such tight-junction molecules are involved in functions other than the "fence" and "barrier" roles of zonulae occludentes
    Type of Publication: Journal article published
    PubMed ID: 16372193
    Signatur Availability
    BibTip Others were also interested in ...
  • 9
    Keywords: PROTEIN ; TISSUE ; TUMORS ; MONOCLONAL-ANTIBODIES ; DESMOSOMAL PLAQUE ; RIGHT-VENTRICULAR CARDIOMYOPATHY ; ADHERENS JUNCTIONS ; EPITHELIAL DIFFERENTIATION ; adhering junctions ; HEART-MUSCLE CELLS ; plakophilin-2 ; AREA-COMPOSITA ; CYTOSKELETAL ARCHITECTURE ; PROTEIN PLAKOPHILIN-2 ; Myxomata ; Cardiac tumors ; Nuclear plakophilins
    Abstract: Within the characteristic ensemble of desmosomal plaque proteins, the protein plakophilin-2 (Pkp2) is known as a particularly important regulatory component in the cytoplasmic plaques of various other cell-cell junctions, such as the composite junctions () of the myocardiac intercalated disks and in the variously-sized and -shaped complex junctions of permanent cell culture lines derived therefrom. In addition, Pkp2 has been detected in certain protein complexes in the nucleoplasm of diverse kinds of cells. Using a novel set of highly sensitive and specific antibodies, both kinds of Pkp2, the junctional plaque-bound and the nuclear ones, can also be localized to the cytoplasmic plaques of diverse non-desmosomal cell-cell junction structures. These are not only the and the connecting various types of highly proliferative non-epithelial cells growing in culture but also some very proliferative states of cardiac interstitial cells and cardiac myxomata, including tumors growing in situ as well as fetal stages of heart development and cultures of valvular interstitial cells. Possible functions and assembly mechanisms of such Pkp2-positive cell-cell junctions as well as medical consequences are discussed
    Type of Publication: Journal article published
    PubMed ID: 22281687
    Signatur Availability
    BibTip Others were also interested in ...
  • 10
    Keywords: NETWORKS ; ALZHEIMERS-DISEASE ; TRANSGENIC MICE ; MOUSE MODEL ; ASTROCYTES ; VASCULAR NORMALIZATION ; metastasis formation ; CALCIUM WAVES ; IMAGING REVEALS ; 3 DIMENSIONS
    Abstract: A complex and reciprocal communication of cells with each other and with relevant parts of the tissue stroma governs many biological processes in both health and disease. However, in the past, the study of these anatomical and molecular interactions has suffered from a lack of appropriate experimental models. An imaging methodology aimed at changing this should allow intravital display and quantification in an intact non-traumatized organ, imaging over a wide range of time spans including extended periods (i.e., months), many repetitive measurements of the same cell or area to permit the study of the cause and consequence of biological processes, the display of various cell types and their reciprocal interaction with each other in three dimensions, the co-registration of relevant physiological parameters and reporters for selected molecular pathways and as high as possible resolution to visualize sub-cellular structures such as organelles. Remarkably, intravital multiphoton microscopy (in vivo MPLSM) through a chronic cranial window allows us to do all these things, making the brain the inner organ of choice for this technology. Here, we give an overview of the application of in vivo MPLSM to study the choreography of cellular, vascular and molecular interactions in the healthy brain and in neurological diseases. We focus on brain tumor formation, progression and response to therapies. This review further aims at demonstrating that we stand at the beginning of full exploitation of the opportunities provided by this technology and gives clues to future directions that appear most promising.
    Type of Publication: Journal article published
    Signatur Availability
    BibTip Others were also interested in ...
Close ⊗
This website uses cookies and the analysis tool Matomo. More information can be found here...