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  • 1
    Publication Date: 2014-04-15
    Repository Name: EPIC Alfred Wegener Institut
    Type: Inbook , NonPeerReviewed
    Format: application/pdf
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  • 2
    Publication Date: 2011-08-24
    Description: A magnetic tweezer was constructed to apply controlled tensional forces (10 pN to greater than 1 nN) to transmembrane receptors via bound ligand-coated microbeadswhile optically measuring lateral bead displacements within individual cells. Use of this system with wild-type F9 embryonic carcinoma cells and cells from a vinculin knockout mouse F9 Vin (-/-) revealed much larger differences in the stiffness of the transmembrane integrin linkages to the cytoskeleton than previously reported using related techniques that measured average mechanical properties of large cell populations. The mechanical properties measured varied widely among cells, exhibiting an approximately log-normal distribution. The median lateral bead displacement was 2-fold larger in F9 Vin (-/-) cells compared to wild-type cells whereas the arithmetic mean displacement only increased by 37%. We conclude that vinculin serves a greater mechanical role in cells than previously reported and that this magnetic tweezer device may be useful for probing the molecular basis of cell mechanics within single cells. Copyright 2000 Academic Press.
    Keywords: Life Sciences (General)
    Type: Biochemical and biophysical research communications (ISSN 0006-291X); Volume 277; 1; 93-9
    Format: text
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  • 3
    Publication Date: 2019-07-13
    Description: A magnetic twisting stimulator was developed based on the previously published technique of magnetic twisting cytometry. Using ligand-coated ferromagnetic microbeads, this device can apply mechanical stresses with varying amplitudes, duration, frequencies, and waveforms to specific cell surface receptors. Biochemical and biological responses of the cells to the mechanical stimulation can be assayed. Twisting integrin receptors with RGD (Arg-Gly-Asp)-containing peptide-coated beads increased endothelin-1 (ET-1) gene expression by 〉100%. In contrast, twisting scavenger receptors with acetylated low-density lipoprotein-coated beads or twisting HLA antigen with anti-HLA antibody-coated beads did not lead to alterations in ET-1 gene expression. In situ hybridization showed that the increase in ET-1 mRNA was localized in the cells that were stressed with the RGD-coated beads. Blocking stretch-activated ion channels with gadolinium, chelating Ca2+ with EGTA, or inhibiting tyrosine phosphorylation with genistein abolished twist-induced ET-1 mRNA elevation. Abolishing cytoskeletal tension with an inhibitor of the myosin ATPase, with an inhibitor of myosin light chain kinase, or with an actin microfilament disrupter blocked twisted-induced increases in ET-1 expression. Our results are consistent with the hypothesis that the molecular structural linkage of integrin-cytoskeleton is an important pathway for stress-induced ET-1 gene expression.
    Keywords: Life Sciences (General)
    Type: American journal of physiology. Cell physiology (ISSN 0363-6143); 280; 6; C1475-84
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  • 4
    Publication Date: 2019-07-13
    Description: This study was carried out to discriminate between two alternative hypotheses as to how cells sense mechanical forces and transduce them into changes in gene transcription. Do cells sense mechanical signals through generalized membrane distortion or through specific transmembrane receptors, such as integrins? Here we show that mechanical stresses applied to the cell surface alter the cyclic AMP signalling cascade and downstream gene transcription by modulating local release of signals generated by activated integrin receptors in a G-protein-dependent manner, whereas distortion of integrins in the absence of receptor occupancy has no effect.
    Keywords: Life Sciences (General)
    Type: Nature cell biology (ISSN 1465-7392); 2; 9; 666-8
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  • 5
    Publication Date: 2019-07-17
    Description: The spontaneous motion of microbeads bound to the cytoskeleton of living cells is not an ordinary random walk. Unlike Brownian motion, the mean-square displacement undergoes a transition from subdiffusive to superdiffusive behavior with time. This transition is associated with characteristic changes of the turning angle distribution. Recent experimental data demonstrated that force fluctuations measured in an elastic hydrogel matrix beneath the cell correlate with the bead motion [C. Raupach, Phys. Rev. E 76, 011918 (2007)]. These data indicate that the bead trajectory is driven by motor forces originating from the actomyosin network and that cytoskeletal remodeling processes with short- and long-time dynamics are mainly responsible for the non-Brownian behavior. We show that the essential statistical properties of the spontaneous bead motion can be reproduced by a particle diffusing in a potential well with a slowly drifting minimum position. Based on this simple model, which can be solved analytically, we develop a biologically plausible numerical model of a tensed and continuously remodeling actomyosin network that accounts quantitatively for the measured data.
    Repository Name: EPIC Alfred Wegener Institut
    Type: Article , NonPeerReviewed
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  • 6
    Publication Date: 2019-07-17
    Repository Name: EPIC Alfred Wegener Institut
    Type: Article , NonPeerReviewed
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  • 7
    Publication Date: 2019-07-17
    Repository Name: EPIC Alfred Wegener Institut
    Type: Conference , NonPeerReviewed
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  • 8
    Publication Date: 2019-07-17
    Repository Name: EPIC Alfred Wegener Institut
    Type: Conference , NonPeerReviewed
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  • 9
    Publication Date: 2019-07-17
    Description: Cytoskeletal (CSK) dynamics such as remodeling and reorganization can be studied by tracking the spontaneous motion of CSK-bound particles. Particle motion is thought to be driven by local, ATP-dependent intracellular force fluctuations due to polymerization processes and motor proteins, and to be impeded by a viscoelastic, metastable cytoskeletal network. The mechanisms that link particle motion to force fluctuations and the CSK dynamics remain unclear. We report simultaneous measurements of the spontaneous motion of CSK-bound particles and of cellular force fluctuations. Cellular force fluctuations were measured by tracking fluorescent markers embedded in an elastic polyacrylamide hydrogel substrate that served as an extracellular matrix (ECM). The motion of CSK-bound particles and markers embedded in the ECM showed both persistence and superdiffusive behavior. Moreover, the movements of CSK-bound beads were temporally and spatially correlated with force fluctuations in the ECM. The findings suggest that the spontaneous motion of CSK-bound beads is driven not by random, local stress fluctuations within a viscoelastic continuum or cage, but rather by stress fluctuations within a tensed and constantly remodeling CSK network that transmits stresses over considerable distances to the ECM.
    Repository Name: EPIC Alfred Wegener Institut
    Type: Article , NonPeerReviewed
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  • 10
    Publication Date: 2019-07-17
    Repository Name: EPIC Alfred Wegener Institut
    Type: Article , NonPeerReviewed
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