Search results for "intermediate filament"
showing 10 items of 103 documents
Desmosomes: interconnected calcium-dependent structures of remarkable stability with significant integral membrane protein turnover
2002
Desmosomes are prominent cell adhesion structures that are major stabilizing elements, together with the attached cytoskeletal intermediate filament network, of the cytokeratin type in epithelial tissues. To examine desmosome dynamics in tightly coupled cells and in situations of decreased adhesion, fluorescent desmosomal cadherin desmocollin 2a (Dsc2a) chimeras were stably expressed in human hepatocellular carcinoma-derived PLC cells (clone PDc-13) and in Madin-Darby canine kidney cells (clone MDc-2) for the continuous monitoring of desmosomes in living cells. The hybrid polypeptides integrated specifically and without disturbance into normal-appearing desmosomes that occurred in associati…
Electron Microscopic Contrast of the Cytoskeleton and Junctional Complexes of Intestinal Epithelial Cells by Ethanolic Phosphotungstic Acid
2000
After glutaraldehyde fixation and treatment with ethanolic phosphotungstic acid (E-PTA) before plastic embedding, sections of rat large intestine showed a characteristic electron contrasting pattern in epithelial cells. The axis of microvilli, terminal web, a thin band below the luminal plasma membrane, centrioles and junctional complexes (tight junctions, adherens junctions, and desmosomes) appeared highly contrasted. In addition to protein components of microfilaments and intermediate filaments, proteins from the junctional complexes could also be implicated in the contrasting reaction with E-PTA. Mitochondrial membranes, chromatin masses, and nucleoli of enterocytes showed considerable e…
Induction of rapid and reversible cytokeratin filament network remodeling by inhibition of tyrosine phosphatases
2002
The cytokeratin filament network is intrinsically dynamic, continuously exchanging subunits over its entire surface, while conferring structural stability on epithelial cells. However, it is not known how cytokeratin filaments are remodeled in situations where the network is temporarily and spatially restricted. Using the tyrosine phosphatase inhibitor orthovanadate we observed rapid and reversible restructuring in living cells, which may provide the basis for such dynamics. By examining cells stably expressing fluorescent cytokeratin chimeras, we found that cytokeratin filaments were broken down and then formed into granular aggregates within a few minutes of orthovanadate addition. After …
Light-induced resistance of the keratin network to the filament-disrupting tyrosine phosphatase inhibitor orthovanadate.
2003
Epidermal keratinocytes respond to low-dose light irradiation by inducing signaling cascades that lead to long-term effects on gene transcription thereby protecting cells against damage. In contrast, little is known about immediate light-induced alterations of structural proteins. We have made the intriguing observation that light produces fundamental changes in the properties of the keratin filament system of cultured epidermoid A-431 cells. A short light exposure (1–10 min) causes the keratin cytoskeleton to become immediately resistant to the tyrosine phosphatase inhibitor orthovanadate, which otherwise disrupts the keratin filament network completely in just a few minutes. This protecti…
0393: Impact of miR-378* and its target desmin intermediate filament on mitochondria distribution in cardiomyocytes
2014
Background MiR-378 and miR-378* microRNAs are derived from an intron of the PGC-1β gene, a regulator of mitochondrial biogenesis. Their expression is either repressed or increased during heart failure depending on the model. Through proteomics approaches, we previously identified new targets of these miRs in H9c2 fetal cardiomyoblasts, among which lactate dehydrogenase for miR-378 and key cytoskeletal proteins for miR-378*. Aims To better assess its role in energy metabolism and differentiation; we overexpressed miR-378 and miR-378* in primary neonate rat cardiomyocytes (NRC) that are more differentiated and less proliferative than H9c2 cardiomyoblasts. Results We identified desmin as a new…
In vivo imaging and quantification of the continuous keratin filament network turnover
2008
Keratin polypeptides are major components of the epithelial cytoskeleton forming a filamentous 3D-network. Like intermediate filament polypeptides of other cell types, keratins make up a stable, but elastic network that is responsible for mechanical stress resilience. At the same time the keratin network is able to change its shape during development, cell division, metastasis and cell migration.
Identification of Novel Principles of Keratin Filament Network Turnover in Living Cells
2004
It is generally assumed that turnover of the keratin filament system occurs by exchange of subunits along its entire length throughout the cytoplasm. We now present evidence that a circumscribed submembranous compartment is actually the main site for network replenishment. This conclusion is based on the following observations in living cells synthesizing fluorescent keratin polypeptides: 1) Small keratin granules originate in close proximity to the plasma membrane and move toward the cell center in a continuous motion while elongating into flexible rod-like fragments that fuse with each other and integrate into the peripheral KF network. 2) Recurrence of fluorescence after photobleaching i…
Imaging of Keratin Dynamics during the Cell Cycle and in Response to Phosphatase Inhibition
2004
Publisher Summary The characterization and development of autofluorescent proteins, most prominently of the green florescent protein, have provided tools to label cellular structures such that they can be examined in living cells. This chapter highlights the potential of live cell imaging in providing novel and unprecedented insights into the dynamic organization of the keratin cytoskeleton and outlines the important aspects of this method. The live cell imaging experiments suggest that the driving force behind the vectorial and dynamic keratin distribution patterns relies both on microtubules and microfilaments and their associated factors. The studies on the dynamics of the keratin cytosk…
Multipotential nestin and Isl-1 positive mesenchymal stem cells isolated from human pancreatic islets.
2006
Mesenchymal cells in the developing pancreas express the neural stem cell marker nestin and the transcription factor islet-1 (Isl-1). Using defined culture conditions we isolated on a single cell basis nestin producing cells from human pancreatic islets. These cells were immortalized with lentiviral vectors coding for telomerase and mBmi. They are positive for Isl-1 and nestin and have the potential to adopt a pancreatic endocrine phenotype with expression of critical transcription factors including Ipf-1, Isl-1, Ngn-3, Pax4, Pax6, Nkx2.2, and Nkx6.1 as well as the islet hormones insulin, glucagon, and somatostatin. In addition, they can be differentiated into human albumin producing cells …
Transgenic overexpression of corticotropin releasing hormone provides partial protection against neurodegeneration in an in vivo model of acute excit…
2008
Abstract Corticotropin releasing hormone (CRH) is the central modulator of the mammalian hypothalamic–pituitary–adrenal (HPA) axis. In addition, CRH affects other processes in the brain including learning, memory, and synaptic plasticity. Moreover, CRH has been shown to play a role in nerve cell survival under apoptotic conditions and to serve as an endogenous neuroprotectant in vitro . Employing mice overexpressing murine CRH in the CNS, we observed a differential response of CRH-overexpressing mice (CRH-COE hom -Nes) to acute excitotoxic stress induced by kainate compared with controls (CRH-COE con -Nes). Interestingly, CRH-overexpression reduced the duration of epileptic seizures and pre…