Developmental loss of laminin from the interstitial extracellular matrix correlates with decreased laminin gene expression

Development ◽  
1990 ◽  
Vol 110 (4) ◽  
pp. 1285-1293 ◽  
Author(s):  
A. Kucherer-Ehret ◽  
J. Pottgiesser ◽  
G.W. Kreutzberg ◽  
H. Thoenen ◽  
D. Edgar
Keyword(s):  
Extracellular Matrix ◽  
Ultrastructural Level ◽  
Basement Membranes ◽  
Mrna Levels ◽  
Physiological Conditions ◽  
Subsequent Decrease ◽  
Mouse Tissues ◽  
Subunit Expression ◽  
A Chain ◽  
Almost All

The expression of the polypeptide subunits of the glycoprotein laminin in developing mouse tissues was analysed by immunoblots and Northern blots, and by immunohistochemistry at the ultrastructural level. In the neonate, almost all the laminin of the sciatic nerve was freely extractable and was located mainly in the mesenchymal interstitial extracellular matrix, rather than in basement membranes. During the first two postnatal weeks, the distribution of laminin shifted to assume the adult pattern, most being located in basement membranes and insoluble under physiological conditions. Analysis of laminin subunit expression showed that both the mRNA for the laminin B chains and the corresponding polypeptides are widely expressed in nerve and other tissues, the mRNA levels decreasing during the first two postnatal weeks as the amount of laminin in the tissue increased. In contrast, the A chain mRNA and polypeptide were undetectable in nerve at any age studied, although they were present in perinatal kidney and placenta. It is proposed that the large amount of soluble laminin present in the developing interstitial extracellular matrix is a consequence of the high levels of expression of laminin mRNA, the subsequent decrease in expression resulting in the adult distribution where most laminin is insoluble within the basement membrane.

scholarly journals Extracellular Matrix Remodeling in the Retina and Optic Nerve of a Novel Glaucoma Mouse Model

Biology ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 169
Author(s):  
Jacqueline Reinhard ◽  
Susanne Wiemann ◽  
Sebastian Hildebrandt ◽  
Andreas Faissner
Keyword(s):  
Extracellular Matrix ◽  
Optic Nerve ◽  
Protein Tyrosine Phosphatase ◽  
Tyrosine Phosphatase ◽  
Nerve Fibers ◽  
Receptor Protein ◽  
Mrna Levels ◽  
Protein Tyrosine ◽  
Tenascin C ◽  
Iop Elevation

Glaucoma is a neurodegenerative disease that is characterized by the loss of retinal ganglion cells (RGC) and optic nerve fibers. Increased age and intraocular pressure (IOP) elevation are the main risk factors for developing glaucoma. Mice that are heterozygous (HET) for the mega-karyocyte protein tyrosine phosphatase 2 (PTP-Meg2) show chronic and progressive IOP elevation, severe RGCs loss, and optic nerve damage, and represent a valuable model for IOP-dependent primary open-angle glaucoma (POAG). Previously, evidence accumulated suggesting that glaucomatous neurodegeneration is associated with the extensive remodeling of extracellular matrix (ECM) molecules. Unfortunately, little is known about the exact ECM changes in the glaucomatous retina and optic nerve. Hence, the goal of the present study was to comparatively explore ECM alterations in glaucomatous PTP-Meg2 HET and control wild type (WT) mice. Due to their potential relevance in glaucomatous neurodegeneration, we specifically analyzed the expression pattern of the ECM glycoproteins fibronectin, laminin, tenascin-C, and tenascin-R as well as the proteoglycans aggrecan, brevican, and members of the receptor protein tyrosine phosphatase beta/zeta (RPTPβ/ζ) family. The analyses were carried out in the retina and optic nerve of glaucomatous PTP-Meg2 HET and WT mice using quantitative real-time PCR (RT-qPCR), immunohistochemistry, and Western blot. Interestingly, we observed increased fibronectin and laminin levels in the glaucomatous HET retina and optic nerve compared to the WT group. RT-qPCR analyses of the laminins α4, β2 and γ3 showed an altered isoform-specific regulation in the HET retina and optic nerve. In addition, an upregulation of tenascin-C and its interaction partner RPTPβ/ζ/phosphacan was found in glaucomatous tissue. However, comparable protein and mRNA levels for tenascin-R as well as aggrecan and brevican were observed in both groups. Overall, our study showed a remodeling of various ECM components in the glaucomatous retina and optic nerve of PTP-Meg2 HET mice. This dysregulation could be responsible for pathological processes such as neovascularization, inflammation, and reactive gliosis in glaucomatous neurodegeneration.

scholarly journals Mineral Element Deposition and Gene Expression across Different Tissues of Cherry Valley Ducks

Animals ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 238
Author(s):  
Qianqian Song ◽  
Yi Zhang ◽  
Hao Bai ◽  
Li Zhong ◽  
Xiaofan Li ◽  
...  
Keyword(s):  
Significant Negative Correlation ◽  
Mineral Elements ◽  
Breast Muscle ◽  
Thigh Muscle ◽  
Mrna Levels ◽  
High Quality ◽  
Atomic Fluorescence ◽  
Breeding Efficiency ◽  
Almost All ◽  
Different Tissues

This study was conducted to investigate the deposition of several mineral elements and the mRNA levels of mineral-related genes across different tissues of cherry valley ducks. The contents of magnesium (Mg), potassium (K), zinc (Zn), and selenium (Se) in ducks’ breast muscle, thigh muscle, liver, skin, and tibia at the age of 0, 21, 35, 49, and 63 days, respectively, were measured using an atomic fluorescence spectrophotometer, while the mRNA levels of mineral-related genes were detected by qRT-PCR. The results revealed that the dynamics of Mg and K were generally similar in each tissue, with a significant positive correlation (p < 0.05). In the breast muscle, thigh muscle, and liver, the contents of almost all mineral elements reached their peak values (p < 0.05) at the age of 49 to 63 days. Interestingly, the expression of most mineral-related genes was the highest at birth (p < 0.05). In addition, there was a significant negative correlation between the expression of ATP1A1 and the deposition of K (r = −0.957, p < 0.05), and a similar result was found for the expression of ATP8 and the deposition of Zn (r = −0.905, p < 0.05). Taken together, Mg and K could be used as joint indicators for the precise breeding of the high-quality strain of cherry valley ducks, while the age of 49 to 63 days could be used as the reference for the best marketing age. In addition, ATP1A1 and ATP8 could be used as the key genes to detect K and Zn, respectively. Hence, the findings of this study can be used to improve the production and breeding efficiency of high-quality meat ducks.

Global gene expression profiles reveal an increase in mRNA levels of collagens, MMPs, and TIMPs in late radiation enteritis

2004 ◽  
Vol 287 (4) ◽  
pp. G875-G885 ◽  
Author(s):  
Carine Strup-Perrot ◽  
Denis Mathé ◽  
Christine Linard ◽  
Dominique Violot ◽  
Fabien Milliat ◽  
...  
Keyword(s):  
Gene Expression ◽  
Extracellular Matrix ◽  
Expression Profiles ◽  
Cdna Array ◽  
Muscularis Propria ◽  
Gelatin Zymography ◽  
Tissue Inhibitors Of Metalloproteinases ◽  
Radiation Enteritis ◽  
Mrna Levels ◽  
Fixed Tissue

Radiation enteritis, a common complication of radiation therapy for abdominal and pelvic cancers, is characterized by severe transmural fibrosis associated with mesenchymal cell activation, tissue disorganization, and deposition of fibrillar collagen. To investigate the mechanisms involved in this pathological accumulation of extracellular matrix, we studied gene expression of matrix components along with that of genes involved in matrix remodeling, matrix metalloproteinases (MMPs), and tissue inhibitors of metalloproteinases (TIMPs). Hybrid selection on high-density cDNA array, real-time RT-PCR, gelatin zymography and imunohistochemistry were used to characterize the mRNA expression profile, activity, and tissue location of extracellular matrix-related genes in radiation enteritis compared with healthy ileum. cDNA array analysis revealed a strong induction of genes coding for collagens I, III, IV, VI, and VIII, SPARC, and tenascin-C, extracellular-matrix degrading enzymes (MMP-1, -2, -3, -14, -18+19), and metalloproteinase inhibitors (TIMP-1, -2, plasminogen activator inhibitor-1) in radiation enteritis. This increase was correlated with the degree of infiltration of the mucosa by inflammatory cells, and the presence of differentiated mesenchymal cells in the submucosa and muscularis propria. Despite the fact that expression of collagens, MMPs, and TIMPs simultaneously increase, quantification of net collagen deposition shows an overall accumulation of collagen. Our results indicate that late radiation enteritis tissues are subjected to active process of fibrogenesis as well as fibrolysis, with a balance toward fibrogenesis. This demonstrates that established fibrotic tissue is not scarred fixed tissue but is subjected to a dynamic remodeling process.

Analisis Sistem Kelistrikan pada Beban Motor Mesin Pencacah Sampah Plastik

2021 ◽  
Vol 2 (1) ◽  
Author(s):  
Yanolanda Suzantry Handayani ◽  
Junas Haidi ◽  
Agun Mardian
Keyword(s):  
Single Phase ◽  
Plastic Waste ◽  
Motor Power ◽  
Electric Motors ◽  
Motor Speed ◽  
Speed Measurement ◽  
Waste Load ◽  
A Chain ◽  
Almost All ◽  
Modern Era

In this modern era, the activities of almost all humans depend on machines they make, such as single-phase induction electric motors, which are used to chop plastic waste. This chopping machine aims to help plastic collectors process plastic waste into small pieces, making it easier to pack and ship plastic out of the area for reprocessing. The plastic waste shredding machine is made using a crushing system with a fan-shaped blade construction consisting of 39 blades divided by two rotating rows opposite the cover box using a chain motor gear transmission element. Most of the chopper machines on the market use engines with diesel or diesel fuel, therefore a chopper machine using an electric motor is designed to compare the motor power without the addition of capacitors and capacitors. The waste load used for motors without additional capacitors, medium and large bottles measuring 375 ml to 1500 ml, the machine can chop as much as 800 grams with the highest measurement of power 578.0 Watt, current 4.192 A, the lowest motor speed measurement is 1414 rpm and the reducer speed is 22.9 rpm . The waste load used for motors with additional capacitors, medium and large bottles measuring 375 ml to 1500 ml, the machine can chop 1000 grams with the highest measurement of power 732.7 Watt, current 4.149 A, the lowest motor speed measurement is 1464 rpm and the reducer speed is 22.9 rpm.

The distribution of fibronectin, laminin and entactin in the neurulating rat embryo studied by indirect immunofluorescence

Development ◽  
1986 ◽  
Vol 94 (1) ◽  
pp. 95-112
Author(s):  
Fiona Tuckett ◽  
Gillian M. Morriss-Kay
Keyword(s):  
Extracellular Matrix ◽  
Indirect Immunofluorescence ◽  
Active Sites ◽  
Polyclonal Antibodies ◽  
Tube Formation ◽  
Basement Membranes ◽  
General Distribution ◽  
Rat Embryo ◽  
Structural Element ◽  
The Matrix

This paper forms part of our study of the extracellular matrix and its role in the morphogenesis of the brain during the period of neurulation in the rat embryo. Using indirect immunofluorescence with polyclonal antibodies, we present here a descriptive study of the distribution of the matrix glycoproteins fibronectin, laminin and entactin. The observed distribution of the fibronectin matrix implicates it in providing a structural element in several morphologically active sites; in addition our observations support the previously suggested involvement of fibronectin in the migration of neural crest cells. Entactin was present only in the basement membranes in conjunction with laminin which was not itself confined to these regions. Laminin was also identified within the mesenchymal extracellular matrix, and its general distribution confirms the previously documented role of laminin in maintaining epithelial structure and organization. No patterning in the distribution of these three glycoproteins could be correlated with the change in shape of the neural epithelium associated with either tube formation or neuromere morphogenesis.

Characterization of a factor that stimulates hydrolysis of GTP bound to ras gene product p21 (GTPase-activating protein) and correlation of its activity to cell density

1988 ◽  
Vol 8 (10) ◽  
pp. 4169-4173
Author(s):  
M Hoshino ◽  
M Kawakita ◽  
S Hattori
Keyword(s):  
Gene Product ◽  
Cell Density ◽  
Cultured Cells ◽  
Specific Activity ◽  
Gtpase Activating Protein ◽  
Ras Gene ◽  
Cell Extracts ◽  
Mouse Tissues ◽  
Almost All ◽  
Hydrolysis Of

The postmicrosomal fraction of the extract from NIH 3T3 and BALB/c 3T3 cells stimulated the hydrolysis of GTP bound to H-ras gene product p21 by severalfold. The stimulation was observed with normal p21 but not with p21 with valine as the 12th residue. This specificity is similar to that of GTPase-activating protein (GAP) for N-ras p21 described by M. Trahey and F. McCormick (Science 238:542-545, 1987). Consistent with this specificity, analysis of p21-bound nucleotides in living cells revealed that almost all normal p21 bound GDP, whereas oncogenic mutant p21s bound both GTP and GDP. Similar activity was also found in various mouse tissues, with brain tissue showing the highest specific activity. When cell extracts were prepared from cultured cells, there was a linear relationship between GAP activity and cell density. These results suggest the factor is involved in the regulation of cell proliferation.

Increased extracellular matrix synthesis and mRNA in mesangial cells grown in high-glucose medium

1991 ◽  
Vol 260 (2) ◽  
pp. F185-F191 ◽  
Author(s):  
S. H. Ayo ◽  
R. A. Radnik ◽  
W. F. Glass ◽  
J. A. Garoni ◽  
E. R. Rampt ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Protein Synthesis ◽  
High Glucose ◽  
Mesangial Cells ◽  
Matrix Proteins ◽  
Cell Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mrna Levels ◽  
Glomerular Mesangial Cells ◽  
Glomerular Mesangium

Nodular expansion of glomerular mesangium with increased amounts of extracellular matrix (ECM) material is pathognomic of diabetic nephropathy. The precise mechanisms involved in this accumulation are unknown. Recently, we reported using a solid-phase enzyme-linked immunosorbent assay (ELISA) technique that glomerular mesangial cells, the principal cell type residing in glomerular mesangium, accumulate 50–60% more fibronectin (FN), laminin (LM), and type IV collagen (T-IV) when cultured in medium containing high glucose (30 mM) (S. H. Ayo, R. A. Rodnik, J. Garoni, W. F. Glass II, and J. I. Kreiberg. Am. J. Pathol. 136: 1339-1348, 1990). ECM assembly is controlled by its rate of synthesis and degradation, as well as its binding and rate of incorporation into the ECM. To elucidate the mechanisms involved, pulse-chase experiments were designed to estimate ECM protein synthesis from the incorporation of Trans-35S [( 35S]methionine, [35S]cysteine) into immunoprecipitated FN, LM, and T-IV. mRNA levels were examined, and degradation rates were estimated from the disappearance of radioactivity from matrix proteins in mesangial cells previously incubated with Trans-35S. One week of growth in 30 mM glucose resulted in approximately 40–50% increase in the synthesis of all three matrix proteins compared with 10 mM glucose-grown cells. This was accompanied by a significant increase in the transcripts for all three matrix proteins (approximately twofold). The specific activity of the radiolabel in trichloroacetic acid-precipitable cell protein showed no difference between cells grown in 10 or 30 mM glucose, indicating that total protein synthesis was unchanged. After 1 wk, the rate of FN, LM, and T-IV collagen degradation was unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Emilin, a component of elastic fibers preferentially located at the elastin-microfibrils interface.

1993 ◽  
Vol 121 (1) ◽  
pp. 201-212 ◽  
Author(s):  
G M Bressan ◽  
D Daga-Gordini ◽  
A Colombatti ◽  
I Castellani ◽  
V Marigo ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Chick Embryo ◽  
Culture Medium ◽  
Close Contact ◽  
Corneal Stroma ◽  
Ultrastructural Level ◽  
Elastic Fibers ◽  
Specific Antibodies ◽  
Gold Particles ◽  
Immunofluorescence Studies

The fine distribution of the extracellular matrix glycoprotein emilin (previously known as glycoprotein gp115) (Bressan, G. M., I. Castellani, A. Colombatti, and D. Volpin. 1983. J. Biol. Chem. 258: 13262-13267) has been studied at the ultrastructural level with specific antibodies. In newborn chick aorta the protein was exclusively found within elastic fibers. In both post- and pre-embedding immunolabeling emilin was mainly associated with regions where elastin and microfibrils are in close contact, such as the periphery of the fibers. This localization of emilin in aorta has been confirmed by quantitative evaluation of the distribution of gold particles within elastic fibers. In other tissues, besides being associated with typical elastic fibers, staining for emilin was found in structures lacking amorphous elastin, but where the presence of tropoelastin has been demonstrated by immunoelectron microscopy. This was particularly evident in the oxitalan fibers of the corneal stroma, in the Descemet's membrane, and in the ciliary zonule. Analysis of embryonic aorta revealed the presence of emilin at early stages of elastogenesis, before the appearance of amorphous elastin. Immunofluorescence studies have shown that emilin produced by chick embryo aorta cells in culture is strictly associated with elastin and that the process of elastin deposition is severely altered by the presence of antiemilin antibodies in the culture medium. The name of the protein was derived from its localization at sites where elastin and microfibrils are in proximity (emilin, elastin microfibril interface located protein).

scholarly journals Monoclonal antibodies to laminin reveal the heterogeneity of basement membranes in the developing and adult mouse tissues.

1984 ◽  
Vol 98 (3) ◽  
pp. 971-979 ◽  
Author(s):  
Y J Wan ◽  
T C Wu ◽  
A E Chung ◽  
I Damjanov
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Adult Mouse ◽  
Basement Membranes ◽  
Newborn Mouse ◽  
Preimplantation Stage ◽  
Mouse Tissues ◽  
Reichert's Membrane ◽  
Anatomic Locations ◽  
Immunohistochemical Studies

Two monoclonal antibodies raised against laminin isolated from a mouse parietal yolk sac cell line were used for immunohistochemical studies of basement membranes of the mouse embryo and various fetal and adult tissues. No immunoreactivity with either of the two monoclonal antibodies could be detected in the preimplantation-stage embryos, although it has been shown that these embryos contain extracellular laminin reactive with the conventional polyclonal antilaminin antibodies. Reichert's membrane in early postimplantation stages of development reacted with the monoclonal antibody LAM-I but not with the antibody LAM-II. However, from day 8 of pregnancy onward the Reichert's membrane reacted with both antibodies. Basement membranes of the embryo proper were unreactive with both monoclonal antibodies until day 12 of pregnancy. By day 14 some basement membranes of the fetal tissues became reactive with one or both monoclonal antibodies, whereas others remained still unreactive. In the 17-d fetus and the newborn mouse most of the basement membranes reacted with both monoclonal antibodies, whereas others still reacted with only one. Similar heterogeneity in the immunoreactivity of basement membranes of various tissues was noted in the adult mouse as well. These results indicate that the immunoreactivity of laminin in the extracellular matrix changes during development and that the basement membranes in various anatomic locations display heterogeneity even in the adult mouse.

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