Home About us Contact
|
Home About us Contact | ||
|
|
||
Matrix Assembly (matrix + assembly)
Selected AbstractsEXTRACELLULAR MATRIX ASSEMBLY IN DIATOMS (BACILLARIOPHYCEAE).JOURNAL OF PHYCOLOGY, Issue 2 2006The effects of phosphate (P) limitation, varying salinity (5,65 psu), and solid media growth conditions on the polysaccharides produced by the model diatom, Phaeodactylum tricornutum Bohlin were determined. Sequential extraction was used to separate polymers into colloidal (CL), colloidal extracellular polymeric substances (cEPS), hot water soluble (HW), hot bicarbonate soluble (HB), and hot alkali (HA) soluble fractions. Media-soluble polymers (CL and cEPS) were enriched in 4-linked mannosyl, glucosyl, and galactosyl residues as well as terminal and 3-linked xylosyl residues, whereas HW polymers consisted mainly of 3-linked glucosyl as well as terminal and 2,4-linked glucuronosyl residues. The HB fraction was enriched in terminal and 2-linked rhamnosyl residues derived from the mucilage coating solubilized by this treatment. Hot alkali treatment resulted in the complete dissolution of the frustule releasing 2,3- and 3-linked mannosyl residues. The fusiform morphotype predominated in standard and P-limited cultures and cultures subjected to salinity variations, but growth on solid media resulted in an enrichment of the oval morphotype. The proportion and linkages of 15 residues, including neutral, uronic acid, and O -methylated sugars, varied with environmental conditions. P limitation and salinity changes resulted in 1.5- to 2.5,fold increase in carbohydrate production, with enrichment of highly branched/substituted and terminal rhamnose, xylose, and fucose as well as O -methylated sugars, uronic acids, and sulfate. The increased deoxy- and O -methylated sugar content under unfavorable environments enhances the hydrophobicity of the polymers, whereas the anionic components may play important roles in ionic cross-linking, suggesting that these changes could ameliorate the effects of salinity or P-stress and that these altered polysaccharide characteristics may be useful as bioindicators for environmental stress. [source] Myosin-mediated cytoskeleton contraction and Rho GTPases regulate laminin-5 matrix assemblyCYTOSKELETON, Issue 2 2004Gregory W. deHart Abstract Laminin-5 is a major structural element of epithelial tissue basement membranes. In the matrix of cultured epithelial cells, laminin-5 is arranged into intricate patterns. Here we tested a hypothesis that myosin II-mediated actin contraction is necessary for the proper assembly of a laminin-5 matrix by cultured SCC12 epithelial cells. To do so, the cells were treated with ML-7, a myosin II light chain kinase inhibitor, or Y-27632, an inhibitor of Rho-kinase (ROCK), both of which block actomyosin contraction. Under these conditions, laminin-5 shows an aberrant localization in dense patches at the cell periphery. Since ROCK activity is regulated by the small GTPase Rho, this suggests that members of the Rho family of GTPases may also be important for laminin-5 matrix assembly by SCC12 cells. We confirmed this hypothesis since SCC12 cells expressing mutant proteins that inhibit RhoA, Rac, and Cdc42 assemble the same aberrant laminin-5 protein arrays as drug-treated cells. We have also evaluated the organization of the laminin-5 receptors ,3,1 and ,6,4 integrin and hemidesmosome proteins in ML-7- and Y-27632-treated cells or in cells in which RhoA, Rac, and Cdc42 activity were inhibited. In all instances, ,3,1 and ,6,4 integrin heterodimers, as well as hemidesmosome proteins, localize precisely with laminin-5 in the matrix of the cells. In summary, our results provide evidence that myosin II-mediated actin contraction and the activity of Rho GTPases are necessary for the proper organization of a laminin-5 matrix and localization of hemidesmosome protein arrays in epithelial cells. Cell Motil. Cytoskeleton 57:107,117, 2004. © 2004 Wiley-Liss, Inc. [source] A practical determination strategy of optimal threshold parameter for matrix compression in wavelet BEMINTERNATIONAL JOURNAL FOR NUMERICAL METHODS IN ENGINEERING, Issue 2 2003Kazuhiro Koro Abstract A practical strategy is developed to determine the optimal threshold parameter for wavelet-based boundary element (BE) analysis. The optimal parameter is determined so that the amount of storage (and computational work) is minimized without reducing the accuracy of the BE solution. In the present study, the Beylkin-type truncation scheme is used in the matrix assembly. To avoid unnecessary integration concerning the truncated entries of a coefficient matrix, a priori estimation of the matrix entries is introduced and thus the truncated entries are determined twice: before and after matrix assembly. The optimal threshold parameter is set based on the equilibrium of the truncation and discretization errors. These errors are estimated in the residual sense. For Laplace problems the discretization error is, in particular, indicated with the potential's contribution ,c, to the residual norm ,R, used in error estimation for mesh adaptation. Since the normalized residual norm ,c,/,u, (u: the potential components of BE solution) cannot be computed without main BE analysis, the discretization error is estimated by the approximate expression constructed through subsidiary BE calculation with smaller degree of freedom (DOF). The matrix compression using the proposed optimal threshold parameter enables us to generate a sparse matrix with O(N1+,) (0,,<1) non-zero entries. Although the quasi-optimal memory requirements and complexity are not attained, the compression rate of a few per cent can be achieved for N,1000. Copyright © 2003 John Wiley & Sons, Ltd. [source] Involvement of the cytoskeletal elements in articular cartilage homeostasis and pathologyINTERNATIONAL JOURNAL OF EXPERIMENTAL PATHOLOGY, Issue 1 2009Emma J. Blain Summary The cytoskeleton of all cells is a three-dimensional network comprising actin microfilaments, tubulin microtubules and intermediate filaments. Studies in many cell types have indicated roles for these cytoskeletal proteins in many diverse cellular processes including alteration of cell shape, movement of organelles, migration, endocytosis, secretion, cell division and extracellular matrix assembly. The cytoskeletal networks are highly organized in structure enabling them to fulfil their biological functions. This review will primarily focus on the organization and function of the three major cytoskeletal networks in articular cartilage chondrocytes. Articular cartilage is a major load-bearing tissue of the synovial joint; it is well known that the cytoskeleton acts as a physical interface between the chondrocytes and the extracellular matrix in ,sensing' mechanical stimuli. The effect of mechanical load on cytoskeletal element expression and organization will also be reviewed. Abnormal mechanical load is widely believed to be a risk factor for the development of osteoarthritis. Several studies have intimated that the major cytoskeletal networks are disorganized or often absent in osteoarthritic cartilage chondrocytes. The implications and possible reasoning for this are more widely discussed and placed into context with their potential relevance to disease and therapeutic strategies. [source] Continuous requirement for pp60-Src and phospho-paxillin during fibronectin matrix assembly by transformed cellsJOURNAL OF CELLULAR PHYSIOLOGY, Issue 3 2007Iwona Wierzbicka-Patynowski Fibronectin (FN) matrix assembly is an integrin-mediated process that is regulated by both the extracellular environment and intracellular signaling pathways. The activity of Src-family kinases is important for initiation of FN assembly by normal fibroblasts. Here we report that in HT1080 fibrosarcoma cells, Src kinase activity is required not only for the assembly of FN matrix but also for the maintenance of FN matrix fibrils at the cell surface. Dexamethasone-induced FN fibril formation by these cells was completely blocked for at least 24 h when Src-family kinase activity was inhibited by either PP1 or SU6656. Inhibition of Src after significant matrix had already been assembled, resulted in an increased rate of loss of detergent-insoluble FN. Binding of activation-dependent integrin antibodies reveals a role for Src in maintaining integrin activity. The requirement for Src kinase activity appears to depend, in part, on phosphorylation of paxillin at tyrosine 118 (Y118). Phospho-paxillin co-localized with FN fibrils, and overexpression of GFP-paxillin but not of GFP-paxillinY118F enhanced cell-mediated assembly of FN. Our results indicate that Src maintains FN matrix at the cell surface through its effect on integrin activity and paxillin phosphorylation. J. Cell. Physiol. 210: 750,756, 2007. © 2006 Wiley-Liss, Inc. [source] Non-enzymatic glycation of chondrocyte-seeded collagen gels for cartilage tissue engineeringJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 11 2008Rani Roy Abstract Collagen glycated with ribose (250 mM) in solution (pre-glycation) and as a gel (post-glycation) was seeded with chondrocytes and the effects of glycation on chondrocyte matrix assembly in culture were determined. Pre-glycation enhanced GAG accumulation significantly over controls at both 2 and 4 weeks (p,<,0.05), although at both time points there were no statistical differences in cell number between pre-glycated and control gels. The increased proteoglycan accumulation was shown to be in part due to significantly increased GAG retention by the pre-glycated constructs (p,<,0.05). Total collagen content in these pre-glycated gels was also significantly higher than unglycated gels at 4 weeks (p,<,0.05). With post-glycation of collagen gels, chondrocyte number and GAG accumulation were all significantly lower than controls (p,<,0.05). Post-glycation also inhibited GAG retention by the constructs (p,<,0.05). Given these results, pre-glycation may be an improved processing method for collagen gels for tissue engineering techniques. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 26:1434,1439, 2008 [source] Combined role of type IX collagen and cartilage oligomeric matrix protein in cartilage matrix assembly: Cartilage oligomeric matrix protein counteracts type IX collagen,induced limitation of cartilage collagen fibril growth in mouse chondrocyte culturesARTHRITIS & RHEUMATISM, Issue 12 2009K. Blumbach Objective Defects in the assembly and composition of cartilage extracellular matrix are likely to result in impaired matrix integrity and increased susceptibility to cartilage degeneration. The aim of this study was to determine the functional interaction of the collagen fibril,associated proteins type IX collagen and cartilage oligomeric matrix protein (COMP) during cartilage matrix formation. Methods Primary chondrocytes from mice deficient in type IX collagen and COMP (double-deficient) were cultured in monolayer or alginate beads. Anchorage of matrix proteins, proteoglycan and collagen content, collagen crosslinks, matrix metalloproteinase activity, and mechanical properties of the matrix were measured. Electron microscopy was used to study the formation of fibrillar structures. Results In cartilage lacking both type IX collagen and COMP, matrilin 3 showed decreased matrix anchorage. Less matrilin 3 was deposited in the matrix of double-deficient chondrocytes, while larger amounts were secreted into the medium. Proteoglycans were less well retained in the matrix formed in alginate cultures, while collagen deposition was not significantly affected. Electron microscopy revealed similar cartilage collagen fibril diameters in the cultures of double-deficient and wild-type chondrocytes. In contrast, a larger fibril diameter was observed in the matrix of chondrocytes deficient in only type IX collagen. Conclusion Our results show that type IX collagen and COMP are involved in matrix assembly by mediating the anchorage and regulating the distribution of other matrix macromolecules such as proteoglycans and matrilins and have counteracting effects on collagen fibril growth. Loss of type IX collagen and COMP leads to matrix aberrations that may make cartilage more susceptible to degeneration. [source] | |||||||||||||