scholarly journals In Situ Localization of Gelatinolytic Activity in the Extracellular Matrix of Metastases of Colon Cancer in Rat Liver Using Quenched Fluorogenic DQ-gelatin

2003 ◽  
Vol 51 (6) ◽  
pp. 821-829 ◽  
Author(s):  
Olaf R.F. Mook ◽  
Claudia Van Overbeek ◽  
Eleonora G. Ackema ◽  
Febe Van Maldegem ◽  
Wilma M. Frederiks
Keyword(s):  
Colon Cancer ◽  
Extracellular Matrix ◽  
Rat Liver ◽  
Collagen Type ◽  
Basement Membranes ◽  
Gelatinolytic Activity ◽  
Collagen Type Iv ◽  
Type Iv

Matrix metalloproteinases (MMPs) such as gelatinases are believed to play an important role in invasion and metastasis of cancer. In this study we investigated the possible role of MMP-2 and MMP-9 in an experimental model of colon cancer metastasis in rat liver. We demonstrated with gelatin zymography that the tumors contained MMP-2 and MMP-9, but only MMP-2 was present in the active form. Immunolocalization of MMP-2 showed that the protein was localized at basement membranes of colon cancer cells and in intratumor stroma, associated with extracellular matrix (ECM) components. However, zymography and immunohistochemistry (IHC) do not provide information on the localization of MMP activity. Therefore, we developed an in situ zymography technique using the quenched fluorogenic substrate DQ-gelatin in unfixed cryostat sections. The application of DQ-gelatin in combination with a gelled medium allows precise localization of gelatinolytic activity. Fluorescence due to gelatinolytic activity was found in the ECM of tumors and was localized similarly to both MMP-2 protein and collagen type IV, its natural substrate. The localization of MMP-2 activity and collagen type IV at similar sites suggests a role of MMP-2 in remodeling of ECM of stroma in colon cancer metastases in rat liver.

scholarly journals Expression of collagen type IV in human kidney during prenatal development

2020 ◽  
pp. 111-111
Author(s):  
Vladimir Petrovic ◽  
Ivan Nikolic ◽  
Marko Jovic ◽  
Vladimir Zivkovic ◽  
Miodrag Jocic ◽  
...  
Keyword(s):  
Type Iv Collagen ◽  
Collagen Type ◽  
Kidney Tissue ◽  
Volume Density ◽  
Collagen Iv ◽  
Basement Membranes ◽  
Collagen Type Iv ◽  
Type Iv ◽  
Metanephric Kidney ◽  
Collecting System

Background / Aim. Type IV collagen belongs to the group of non-fibrillar collagens and is an important component of the basement membranes where it accounts for approximately 50% of its structural elements. The aim of the paper was to describe the expression and distribution of collagen type IV in embryonic and fetal metanephric kidney, and to determine the volume density of collagen type IV in kidney tissue in each trimester of development. Methods. The material consisted of 19 human embryos/fetuses, in the gestational age from 8th to 37th week. Kidney tissue specimens were routinely processed to paraffin molds and stained with hematoxylin and eosin and immunohistochemically using polyclonal anti-collagen IV antibody. Stained slides were examined using light microscope and images of the selected areas, under different lens magnification were captured with digital camera. Volume density of collagen type IV was determined by using ImageJ 1.48v and a plugin of the software which inserted a grid system with 336 points. For the data comparison One-Way Analysis of Variance was used. Results. Strong collagen IV immunopositivity was seen in all specimens, with a distribution in the basement membranes of urinary bud, parietal leaf of Bowman?s capsule, glomerular basement membrane, basement membrane of interstitial blood vessels, and basement membranes of nephron tubules and collecting ducts. No statistically significant difference in the volume density of type IV collagen was found between the different trimesters of development. Conclusion. The synthesis and secretion of collagen type IV simultaneously follows the development of nephron structures, collecting system and blood vessels. The volume density of collagen type IV remains constant throughout all the trimesters of metanephric kidney development, indicating that it plays a crucial role in normal development of nephron and collecting system structures, as well as in maintaining the normal kidney function.

scholarly journals Differential expression of parietal epithelial cell and podocyte extracellular matrix proteins in focal segmental glomerulosclerosis and diabetic nephropathy

2019 ◽  
Vol 317 (6) ◽  
pp. F1680-F1694 ◽  
Author(s):  
Gek Cher Chan ◽  
Diana G. Eng ◽  
Jeffrey H. Miner ◽  
Charles E. Alpers ◽  
Kelly Hudkins ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Diabetic Nephropathy ◽  
Epithelial Cell ◽  
Focal Segmental Glomerulosclerosis ◽  
Collagen Type ◽  
Collagen Type Iv ◽  
Type Iv ◽  
Ecm Proteins ◽  
Segmental Glomerulosclerosis ◽  
Parietal Epithelial Cell

In healthy glomeruli, parietal epithelial cell (PEC)-derived extracellular matrix (ECM) proteins include laminin-β1, perlecan, and collagen type IV-α2 and podocyte-specific ECM proteins include laminin-β2, agrin, and collagen type IV-α4. This study aimed to define individual ECM protein isoform expression by PECs in both experimental and human focal segmental glomerulosclerosis (FSGS) and diabetic nephropathy (DN) and to determine if changes were CD44 dependent. In experimental FSGS induced with a cytotoxic podocyte antibody and in the BTBR ob/ob mouse model of DN, PEC-derived protein staining was significantly increased in PECs. Dual staining also showed de novo expression of the podocyte-specific ECM proteins laminin-β2 and agrin in PECs. Similar findings were observed in biopsies from patients with FSGS and DN. Increases in individual ECM proteins colocalized with CD44 in PECs in disease. To determine the role of CD44, FSGS was induced in CD44−/− and CD44+/+ mice. PEC staining for perlecan, collagen type IV-α2, laminin-β2, and agrin were significantly lower in diseased CD44−/− mice compared with diseased CD44+/+ mice. These results show that in experimental and human FSGS and DN, PECs typically in an activated state, produce both PEC-derived and podocyte-specific ECM protein isoforms, and that the majority of these changes were dependent on CD44.

193CHARACTERIZATION OF PORCINE TROPHECTODERM-DERIVED CELLS IN THE PRESENCE OF HBMP4 IN THE ABSENCE OF A FEEDER LAYER

2004 ◽  
Vol 16 (2) ◽  
pp. 218 ◽  
Author(s):  
K.B. Stewart ◽  
A.M. Adams ◽  
S.L. Pratt ◽  
S.L. Stice
Keyword(s):  
Extracellular Matrix ◽  
Cell Growth ◽  
Cell Biology ◽  
Collagen Type ◽  
Growth Characteristic ◽  
Calf Serum ◽  
Embryonic Stem ◽  
Collagen Type Iv ◽  
Type Iv ◽  
Embryonic Disc

A porcine trophoblastic cell line could provide a powerful model for understanding trophoblast cell biology as well as placental gene expression and proteomics in vitro. In this experiment, we derived porcine trophoblastic cells from trophectoderm tissue and assessed their growth on three different extracellular matrix substrates and in three different concentrations of human recombinant bone morphogenetic protein 4 (hBMP4). Human BMP4 has been shown to induce differentiation of human embryonic stem cells into trophoblast lineages. Elongated embryos were flushed using DPBS supplemented with 1% fetal calf serum and penicillin-streptomycin (1X) from the hysterectomized uteri of superovulated and bred prepuberal gilts 15 days post-insemination. The embryonic disc was visualized with a dissecting microscope. The trophectoderm tissue was cut 2–3mm away from the embryonic disc with a scalpel and the trophectoderm tissue was manually dissected into cell aggregates. These aggregates were plated on collagen type IV, Matrigel, and human extracellular matrix (laminin, collagen type IV and heparan sulfate proteoglycan derived from human placenta) in culture medium (DMEM with 15% FCS, 0.1mM 2-mercaptoethanol, 4ngmL−1 basic FGF4 and 1X P/S) in the presence or absence of hBMP4 at 0, 10, or 20ngmL−1. Cell outgrowth was observed within 24 hours of culture. After three days of culture, various cell types (based on size and morphology) were present. Among cultures of predominant large cells were colonies of smaller cells with epithelial-type morphology that had a prominent nucleus and a high nuclear-to-cytoplasmic ratio. The epithelial-type cells grew in tight colonies with definite borders and contained cytoplasmic structures resembling lipid-containing vesicles. These colonies initially appeared on all matrices across all hBMP4 concentrations. After seven days in culture the colonies developed distinct differences across groups. Cell growth on collagen was comprised of tight colonies having definite borders among large cells. Colonies on collagen were larger and more pronounced in both the hBMP4-supplemented groups than when cultured without hBMP4. The Matrigel coated plates contained large sheets of epithelial-type cell growth instead of compact colonies. This type of growth characteristic was present in all hBMP4 treatments on Matrigel. In contrast, few cells survived and propagated on human extracellular matrix. Only small colonies having the desired morphology were among the large cells on human extracellular matrix when cultured in medium containing 10ngmL−1 hBMP4. Cells were passaged and only cells growing on Matrigel could be further cultured. These data suggest that both the cell substrate and hBMP4 affect initial trophoblast outgrowths. Further analysis including immunocytochemistry and RT-PCR is currently being performed to better characterize these cells. Epidemiology/Diseases

Diffuse Leiomyomatosis of the Esophagus: Disorder of Cell-Matrix Interaction?

1998 ◽  
Vol 1 (6) ◽  
pp. 543-549 ◽  
Author(s):  
Paul Thorner ◽  
Laurence Heidet ◽  
Fernando Moreno Merlo ◽  
Vern Edwards ◽  
Corinne Antignac ◽  
...  
Keyword(s):  
Collagen Type ◽  
Rare Condition ◽  
Basement Membranes ◽  
Collagen Type Iv ◽  
Cell Periphery ◽  
Matrix Interaction ◽  
Type Iv ◽  
Cell Matrix ◽  
Reverse Pattern ◽  
Genes Encoding

Diffuse leiomyomatosis (DL) is rare condition characterized by proliferation of smooth muscle in the upper gastrointestinal tract. Most cases are associated with X-linked Alport syndrome and have partial deletions in the genes encoding both the α5 and α6 chains of collagen type IV. We studied aspects of cell-matrix interaction of myocytes in an esophagogastrectomy specimen from a 12-year-old patient with DL. Myocytes had central areas of cytoplasmic rarefaction, which were actin positive and desmin poor, with the reverse pattern of staining at the cell periphery. Electron microscopy (EM) showed that the areas of rarefaction consisted of disorganized aggregates of filaments. The basement membranes ranged from thickened to thinned or absent. Immunohistochemical staining for the α1–α4 chains of collagen type IV, the α1, α2, β2, and γ1 chains of laminin, nidogen, type VI collagen, and fibronectin was normal. There was loss of the α5 and α6 chains of collagen type IV and the β1 chain of laminin. Normal staining for α1, α2, α3, α4, α6, α8, and β1 integrins was noted. Staining for α5 integrin varied from normal to reduced or negative in different cells. In DL, a primary abnormality of basement membrane may be associated with disorganization of the contractile apparatus and alterations of certain integrins. This may reflect a disturbance of cell-matrix interactions that play a role in cell differentiation and internal organization.

scholarly journals Role of myofibroblasts and collagen type IV in patients of IgA nephropathy as markers of renal dysfunction

2010 ◽  
Vol 20 (1) ◽  
pp. 34 ◽  
Author(s):  
RW Minz ◽  
A Bakshi ◽  
S Chhabra ◽  
K Joshi ◽  
V Sakhuja
Keyword(s):  
Renal Dysfunction ◽  
Iga Nephropathy ◽  
Collagen Type ◽  
Collagen Type Iv ◽  
Type Iv

scholarly journals An attempt at quantitation of procollagens I and III and of collagen IV in tooth sections by exposure to the corresponding antibodies followed by 125I-protein A and radioautography.

1980 ◽  
Vol 28 (12) ◽  
pp. 1355-1358 ◽  
Author(s):  
G M Wright ◽  
C P Leblond
Keyword(s):  
Collagen Type ◽  
Protein A ◽  
Collagen Iv ◽  
Basement Membranes ◽  
Collagen Type Iv ◽  
Periodontal Tissue ◽  
Rat Incisor ◽  
Type Iv ◽  
Silver Grains ◽  
Procollagen Iii

The immunoreactivity of procollagen types I and III and of collagen type IV was detected in frozen sections of the growing apical end of rat incisor teeth by an indirect method making use of protein A. The sections were exposed to affinity-purified antibodies against these substances. The bound antibodies were then detected by incubation with radioiodinated protein A, followed by radioautography. This immunoradioautographic approach yielded preparations with low background, in which the reactions could be quantitated by counts of silver grains. The distribution of the radioautographic reactions was essentially the same as that previously observed with direct and indirect peroxidase methods, that is, procollagen I antigenicity predominated in odontoblasts and predentin, with minor amounts in periodontal tissue and pulp; procollagen III antigenicity was present in periodontal tissue and, to a lesser extent, in the pulp; and collagen IV antigenicity was restricted to basement membranes. Moreover, grain counts provided quantitative support for the conclusions on the distribution of procollagen I and III antigenicity.

scholarly journals The role of microtubules in growth cone turning at substrate boundaries.

1995 ◽  
Vol 128 (1) ◽  
pp. 127-137 ◽  
Author(s):  
E Tanaka ◽  
M W Kirschner
Keyword(s):  
Growth Cone ◽  
Collagen Type ◽  
Axon Growth ◽  
Growth Cones ◽  
Collagen Type Iv ◽  
Early Step ◽  
Growth Cone Collapse ◽  
Type Iv ◽  
And Behaviors

To understand the role of microtubules in growth cone turning, we observed fluorescently labeled microtubules in neurons as they encountered a substrate boundary. Neurons growing on a laminin-rich substrate avoided growing onto collagen type IV. Turning growth cones assumed heterogeneous morphologies and behaviors that depended primarily in their extent of adhesion to the substrate. We grouped these behaviors into three categories-sidestepping, motility, and growth-mediated reorientation. In sidestepping and motility-mediated reorientation, the growth cone and parts of the axon were not well attached to the substrate so the acquisition of an adherent lamella caused the entire growth cone to move away from the border and consequently reoriented the axon. In these cases, since the motility of the growth cone dominates its reorientation, the microtubules were passive, and reorientation occurred without significant axon growth. In growth-mediated reorientation, the growth cone and axon were attached to the substrate. In this case, microtubules reoriented within the growth cone to stabilize a lamella. Bundling of the reoriented microtubules was followed by growth cone collapse to form new axon, and further, polarized lamellipodial extension. These observations indicate that when the growth cone remains adherent to the substrate during turning, the reorientation and bundling of microtubules is an important, early step in growth cone turning.

scholarly journals Morphometric analysis of the human endoneurial extracellular matrix components during aging

2021 ◽  
Vol 73 (1) ◽  
pp. 103-110
Author(s):  
Braca Kundalic ◽  
Sladjana Ugrenovic ◽  
Ivan Jovanovic ◽  
Vladimir Petrovic ◽  
Aleksandar Petrovic ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Collagen Type ◽  
Linear Regression Analysis ◽  
Collagen Type I ◽  
Nerve Fibers ◽  
Collagen Type Iv ◽  
Type I ◽  
Type Iv ◽  
Ecm Proteins ◽  
Extracellular Matrix Components

The aim of this study was to analyze the expression of extracellular matrix (ECM) proteins in human endoneurium during aging. We harvested 15 cadaveric sural nerves, distributed in 3 age groups (I: 25-44, II: 45-64, III: 65-86 years old). Histological sections were stained immunohistochemically for the presence of collagen type I, type IV and laminin, and the ImageJ processing program was used in morphometrical analysis to determine the percentages of these endoneurial proteins. In two younger groups, the endoneurial matrix of the sural nerve was composed from about equal proportions of these proteins, which may be considered a favorable microenvironment for the regeneration of nerve fibers. Linear regression analysis showed a significant increase in endoneurial collagen type IV with age, while collagen type I and laminin significantly decreased during the aging process. In cases older than 65 years, remodeling of the endoneurial matrix was observed to be significantly higher for the presence of collagen type IV, and lower for the expression of collagen type I and laminin. This age-related imbalance of ECM proteins could represent a disadvantageous microenvironment for nerve fiber regeneration in older adults. Our findings contribute to the development of therapeutic approaches for peripheral nerve regeneration.

scholarly journals ADAMTS-family protease MIG-17 regulates synaptic allometry by modifying the extracellular matrix and modulating glia morphology during growth

2019 ◽  
Author(s):  
Tingting Ji ◽  
Kai Wang ◽  
Jiale Fan ◽  
Jichang Huang ◽  
Mengqing Wang ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Extracellular Matrix ◽  
Collagen Type ◽  
Collagen Type Iv ◽  
Genetic Screens ◽  
Genetic Studies ◽  
C Elegans ◽  
Type Iv ◽  
Forward Genetic Screens ◽  
Signaling Axis

ABSTRACTSynapses are largely established during embryogenesis and maintained during growth. The mechanisms that regulate synaptic allometry—the maintenance of synaptic positions during growth—are largely unknown. We performed forward genetic screens inC. elegansfor synaptic allometry mutants and identifiedmig-17, a secreted metalloprotease of the conserved ADAMTS family. Through proteomic mass spectrometry analyses, cell biological and genetic studies we determined that MIG-17 is expressed by muscle cells to modulate glia location and morphology. Glia are proximal to synapses, and the glial location and morphology determine synaptic position during growth.Mig-17regulates synapse allometry by influencing epidermal-glia crosstalk through the regulation of basement membrane proteins, including collagen type IV, SPARC and fibulin. Our findings underscore the importance of glia location in the maintenance of synaptic allometry, and uncover a muscle-epidermal-glia signaling axis, mediated through the extracellular matrix, in the regulation of glia morphology and synaptic positions during growth.

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