scholarly journals Giantin is required for intracellular N-terminal processing of type I procollagen

2021 ◽  
Vol 220 (6) ◽  
Author(s):  
Nicola L. Stevenson ◽  
Dylan J.M. Bergen ◽  
Yinhui Lu ◽  
M. Esther Prada-Sanchez ◽  
Karl E. Kadler ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Type I Collagen ◽  
Fish Analysis ◽  
Type I ◽  
Matrix Assembly ◽  
Matrix Deposition ◽  
Type I Procollagen ◽  
Main Protein ◽  
Extracellular Matrix Deposition ◽  
Craniofacial Defects

Knockout of the golgin giantin leads to skeletal and craniofacial defects driven by poorly studied changes in glycosylation and extracellular matrix deposition. Here, we sought to determine how giantin impacts the production of healthy bone tissue by focusing on the main protein component of the osteoid, type I collagen. Giantin mutant zebrafish accumulate multiple spontaneous fractures in their caudal fin, suggesting their bones may be more brittle. Inducing new experimental fractures revealed defects in the mineralization of newly deposited collagen as well as diminished procollagen reporter expression in mutant fish. Analysis of a human giantin knockout cell line expressing a GFP-tagged procollagen showed that procollagen trafficking is independent of giantin. However, our data show that intracellular N-propeptide processing of pro-α1(I) is defective in the absence of giantin. These data demonstrate a conserved role for giantin in collagen biosynthesis and extracellular matrix assembly. Our work also provides evidence of a giantin-dependent pathway for intracellular procollagen processing.

scholarly journals Giantin is required for intracellular N-terminal processing of type I procollagen

2020 ◽  
Author(s):  
Nicola L. Stevenson ◽  
J. M. Bergen Dylan ◽  
Chrissy L. Hammond ◽  
David J. Stephens
Keyword(s):  
Extracellular Matrix ◽  
Type I Collagen ◽  
Fish Analysis ◽  
Type I ◽  
Matrix Assembly ◽  
Matrix Deposition ◽  
Type I Procollagen ◽  
Main Protein ◽  
Extracellular Matrix Deposition ◽  
Craniofacial Defects

AbstractKnockout of the golgin giantin leads to skeletal and craniofacial defects driven by poorly studied changes in glycosylation and extracellular matrix deposition. Here, we sought to determine how giantin impacts the production of healthy bone tissue by focussing on the main protein component of the osteoid, type I collagen. Giantin mutant zebrafish accumulate multiple spontaneous fractures in their caudal fin, suggesting their bones may be more brittle. Inducing new experimental fractures revealed defects in the mineralisation of newly deposited collagen as well as diminished procollagen reporter expression in mutant fish. Analysis of giantin knockout cells expressing a GFP-tagged procollagen showed that procollagen trafficking is independent of giantin. However, our data show that intracellular N-propeptide processing of pro-α1(I) is defective in the absence of giantin. These data demonstrate a conserved role for giantin in collagen biosynthesis and extracellular matrix assembly. Our work also provides evidence of a giantin-dependent pathway for intracellular procollagen processing.

scholarly journals Extracellular Matrix Deposition and Remodeling after Corneal Alkali Burn in Mice

2021 ◽  
Vol 22 (11) ◽  
pp. 5708
Author(s):  
Kazadi N. Mutoji ◽  
Mingxia Sun ◽  
Garrett Elliott ◽  
Isabel Y. Moreno ◽  
Clare Hughes ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Dermatan Sulfate ◽  
Temporal Distribution ◽  
Keratan Sulfate ◽  
Collagen Fibrils ◽  
Type I ◽  
Chemical Injury ◽  
Matrix Deposition ◽  
Alkali Burn ◽  
Extracellular Matrix Deposition

Corneal transparency relies on the precise arrangement and orientation of collagen fibrils, made of mostly Type I and V collagen fibrils and proteoglycans (PGs). PGs are essential for correct collagen fibrillogenesis and maintaining corneal homeostasis. We investigated the spatial and temporal distribution of glycosaminoglycans (GAGs) and PGs after a chemical injury. The chemical composition of chondroitin sulfate (CS)/dermatan sulfate (DS) and heparan sulfate (HS) were characterized in mouse corneas 5 and 14 days after alkali burn (AB), and compared to uninjured corneas. The expression profile and corneal distribution of CS/DSPGs and keratan sulfate (KS) PGs were also analyzed. We found a significant overall increase in CS after AB, with an increase in sulfated forms of CS and a decrease in lesser sulfated forms of CS. Expression of the CSPGs biglycan and versican was increased after AB, while decorin expression was decreased. We also found an increase in KS expression 14 days after AB, with an increase in lumican and mimecan expression, and a decrease in keratocan expression. No significant changes in HS composition were noted after AB. Taken together, our study reveals significant changes in the composition of the extracellular matrix following a corneal chemical injury.

scholarly journals Abnormal Mucosal Extracellular Matrix Deposition Is Associated with Increased TGF-β Receptor-expressing Mesenchymal Cells in a Mouse Model of Colitis

2003 ◽  
Vol 51 (9) ◽  
pp. 1177-1189 ◽  
Author(s):  
Christine V. Whiting ◽  
John F. Tarlton ◽  
Michael Bailey ◽  
Clare L. Morgan ◽  
Paul W. Bland
Keyword(s):  
Extracellular Matrix ◽  
Basement Membrane ◽  
Mouse Model ◽  
Type Iv Collagen ◽  
Type I Collagen ◽  
Transforming Growth Factor ◽  
Mesenchymal Cells ◽  
Type I ◽  
Matrix Deposition ◽  
Type Iv

Transforming growth factor-β (TGF-β) depresses mucosal inflammation and upregulates extracellular matrix (ECM) deposition. We analyzed TGF-β receptors RI and RII as well as ECM components using the CD4+ T-cell-transplanted SCID mouse model of colitis. The principal change in colitis was an increased proportion of TGF-β RII+ mucosal mesenchymal cells, predominantly α-smooth muscle actin (SMA)+ myofibroblasts, co-expressing vimentin and basement membrane proteins, but not type I collagen. TGF-β RII+ SMA− fibroblasts producing type I collagen were also increased, particularly in areas of infiltration and in ulcers. Type IV collagen and laminin were distributed throughout the gut lamina propria in disease but were restricted to the basement membrane in controls. In areas of severe epithelial damage, type IV collagen was lost and increased type I collagen was observed. To examine ECM production by these cells, mucosal mesenchymal cells were isolated. Cultured cells exhibited a similar phenotype and matrix profile to those of in vivo cells. The data suggested that there were at least two populations of mesenchymal cells responsible for ECM synthesis in the mucosa and that ligation of TGF-β receptors on these cells resulted in the disordered and increased ECM production observed in colitic mucosa.

Enhanced assembly of basement membrane matrix by endodermal cells in response to fibronectin substrata

1991 ◽  
Vol 99 (2) ◽  
pp. 443-451
Author(s):  
M.R. Austria ◽  
J.R. Couchman
Keyword(s):  
Extracellular Matrix ◽  
Basement Membrane ◽  
Basement Membranes ◽  
Binding Domain ◽  
Type I ◽  
Matrix Assembly ◽  
Matrix Deposition ◽  
Membrane Matrix ◽  
The Matrix

Basement membranes are complex extracellular matrices contributing to the regulation of growth, migration and differentiation of many cell types. However, little is known about the mechanisms regulating the deposition and assembly of basement membrane from its constituents. We have investigated the role of extracellular matrix molecules in the control of basement membrane matrix assembly by cultured endodermal (PFHR-9) cells. In the presence of fibronectin-depleted serum, substrata of fibronectin or laminin induced an increase in deposition of laminin, type IV collagen and proteoglycans by PFHR-9 cells, in comparison to cells adherent to type I collagen-coated, vitronectin-coated or uncoated substrata. Direct effects of fibronectin or laminin on the degree of cell spreading or rate of proliferation were not responsible for enhanced matrix deposition. The effect did not result from a redirection of basement membrane components to the matrix, since there was no decrease in matrix constituents released to the culture supernatants. Furthermore, the synthesis and release of other molecules that are not basement membrane constituents was unaltered in response to different extracellular matrix substrata. Experiments with fibronectin fragments showed that a 105 × 10(3) Mr ‘cell’-binding domain (containing the cell attachment sequence Arg-Gly-Asp-Ser) was an important contributor to enhanced matrix deposition, while the N-terminal 29 × 10(3) Mr heparin-binding domain also contributed to the effect, particularly with respect to heparan sulfate proteoglycan deposition. It seems that fibronectin has a dual role of action in promoting basement membrane matrix assembly, through direct cell surface interactions, and through the binding of fibronectin to other matrix components that may nucleate or stabilize the matrix assembly.

Preliminary Results of In Vitro Study of Cell Growth in a 45S5 Bioactive Glass as Bone Substitute Using Scanning Electron and Confocal Microscopies

2007 ◽  
Vol 361-363 ◽  
pp. 1111-1114 ◽  
Author(s):  
Rami Maksoud ◽  
Leila Lefebvre ◽  
Laurence Heinrich ◽  
Laurent Gremillard ◽  
Jérôme Chevalier ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Cell Growth ◽  
Confocal Microscopy ◽  
Bioactive Glass ◽  
Bone Substitute ◽  
Type I ◽  
Matrix Deposition ◽  
Extracellular Matrix Components ◽  
Extracellular Matrix Deposition ◽  
Scanning Electron

The aim of this study was to evaluate the cytocompatibility, cell ingrowth and extracellular matrix deposition of a newly developed porous bioactive glass as a bone substitute. Two types of bioactive glass, different in their pore size (75 and 20 ppi, resp. ~350 and ~1200 $m), were used in this study. The materials were seeded with human osteoblastic (MG63) and fibroblastic (M-228 F01 and M-191 F01) cell lines. The cells were visualized by two techniques, scanning electron microscopy and confocal microscopy. For confocal microscopy cell nuclei were labeled with propidium iodide (IP) and the extracellular matrix components (type I collagen and osteocalcin) by specific antibodies. Cells and matrix were visualized by fluorescence. The bioactive glass used in this study was shown to be non cytotoxic. Cell growth and colonization at the surface and in the depth of the material were observed. Extracellular matrix deposition was also demonstrated which proved the proper biofunctionality of the biomaterial. Scanning electron microscope allowed us to visualize cells at a high magnification at the surface of the bioglass and evidenced that the biomaterials were covered by a sheet of cells with their matrix; on the other hand, confocal microscopy permitted us to observe cell ingrowth and matrix deposition within the depth of the substitute. We showed that extracellular matrix was synthesized mainly in the upper levels where the cell population was the most confluent. In summary, this porous bioglass appears promising for bone substitution.

Fat-Soluble Vitamins Affect Composition of Extracellular Matrix Deposited by Human Aortic Smooth Muscle and Endothelial Cells In Vitro

2020 ◽  
Vol 19 (1) ◽  
pp. 36-45
Author(s):  
V. Ivanov ◽  
S. Ivanova ◽  
A. Niedzwiecki ◽  
M. Rath
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Smooth Muscle ◽  
Type Iv Collagen ◽  
Type I ◽  
Aortic Smooth Muscle ◽  
Matrix Deposition ◽  
Type Iv ◽  
Extracellular Matrix Deposition ◽  
Fat Soluble Vitamins

Atherosclerotic cardiovascular disease is accompanied by changes in arterial connective tissue. We evaluated the effects of fat-soluble vitamins A, D, and E individually and in combinations on the composition of extracellular matrix produced and deposited by arterial wall cells, human aortic smooth muscle cells, and endothelial cells. Individually, vitamins D and E stimulated collagen type I extracellular matrix deposition in human aortic smooth muscle cell cultures. However, vitamins A, D, and E reduced collagen type IV deposition by human aortic smooth muscle cell, counteracting the stimulatory effects of vitamin C. The extracellular matrix deposition of heparan sulfate by human aortic smooth muscle cells increased by vitamin C and its combination (C+D+E). β-carotene + D + C induced the extracellular matrix deposition of collagen I by endothelial cells. Vitamin E with other vitamins resulted in either induction (E+C+A) or inhibition (E+D). The extracellular matrix deposition of type IV collagen and elastin by human aortic endothelial cells was not affected by test vitamins, except the extracellular matrix type IV collagen decrease by combinations (A+E), (A+D+E), and (C+D+E). The extracellular matrix deposition of all tested glycosaminoglycans was reduced by vitamin A and its combination (A+C+D+E). Therefore, the fat-soluble vitamins applied individually or in combination—both with each other or with ascorbic acid—can affect extracellular matrix deposition of type I and IV collagens, and key glycosaminoglycans by cultured human aortic arterial wall cells.

scholarly journals The Dynamic Interaction between Extracellular Matrix Remodeling and Breast Tumor Progression

Cells ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 1046
Author(s):  
Jorge Martinez ◽  
Patricio C. Smith
Keyword(s):  
Extracellular Matrix ◽  
Type I Collagen ◽  
Cell Metabolism ◽  
Breast Tumors ◽  
Extracellular Matrix Remodeling ◽  
Type I ◽  
Matrix Remodeling ◽  
Desmoplastic Reaction ◽  
The Impact

Desmoplastic tumors correspond to a unique tissue structure characterized by the abnormal deposition of extracellular matrix. Breast tumors are a typical example of this type of lesion, a property that allows its palpation and early detection. Fibrillar type I collagen is a major component of tumor desmoplasia and its accumulation is causally linked to tumor cell survival and metastasis. For many years, the desmoplastic phenomenon was considered to be a reaction and response of the host tissue against tumor cells and, accordingly, designated as “desmoplastic reaction”. This notion has been challenged in the last decades when desmoplastic tissue was detected in breast tissue in the absence of tumor. This finding suggests that desmoplasia is a preexisting condition that stimulates the development of a malignant phenotype. With this perspective, in the present review, we analyze the role of extracellular matrix remodeling in the development of the desmoplastic response. Importantly, during the discussion, we also analyze the impact of obesity and cell metabolism as critical drivers of tissue remodeling during the development of desmoplasia. New knowledge derived from the dynamic remodeling of the extracellular matrix may lead to novel targets of interest for early diagnosis or therapy in the context of breast tumors.

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