Nonsynchronous changes in myocardial collagen mRNA and protein during aging: effect of DOCA-salt hypertension

1994 ◽  
Vol 267 (6) ◽  
pp. H2237-H2244 ◽  
Author(s):  
S. Besse ◽  
V. Robert ◽  
P. Assayag ◽  
C. Delcayre ◽  
B. Swynghedauw
Keyword(s):  
Aging Effect ◽  
Type I ◽  
Mrna Levels ◽  
Salt Treatment ◽  
Collagen Mrna ◽  
Type I Procollagen ◽  
Salt Hypertension ◽  
Old Rats ◽  
Modify Collagen ◽  
Myocardial Collagen

Myocardial fibrosis has been investigated in 3-, 16-, and 24-mo-old normal rats and also in 24-mo-old rats subjected to deoxycorticosterone acetate (DOCA)-salt treatment-induced-hypertension. Collagen content was assessed both histologically and by hydroxyproline assay. Type I and III procollagen mRNA levels were quantitated by Slot Blot analyses. Aging is associated with fibrosis as shown both biochemically (hydroxyproline concentration in 3-, 16-, and 24-mo-old rats was 0.70 +/- 0.05, 0.92 +/- 0.07, and 1.57 +/- 0.13 mg/g of left ventricle, respectively, P < 0.05 and P < 0.0001 vs. 3 mo) and histologically. By contrast, type I procollagen mRNA levels decreased during aging (from -63%, P < 0.001 in 16-mo-old rats and -51%, P < 0.01 in 24-mo-old rats vs. 3-mo-old rats) as well as type III procollagen mRNA levels. DOCA-salt treatment in 24-mo-old rats had no effect on either the degree of fibrosis or the mRNA levels. We conclude that nonsynchronous changes in myocardial collagen mRNA and protein occur during aging, indicating translational and/or posttranslational mechanisms in collagen regulation. Hypertension during senescence did not modify collagen deposition at either the protein or mRNA levels.

Collagen expression in embryonic chicken chondrocytes treated with phorbol myristate acetate

1985 ◽  
Vol 5 (6) ◽  
pp. 1415-1424
Author(s):  
M H Finer ◽  
L C Gerstenfeld ◽  
D Young ◽  
P Doty ◽  
H Boedtker
Keyword(s):  
Morphological Change ◽  
Type I Collagen ◽  
Late Phase ◽  
Tumor Promoter ◽  
Type I ◽  
Mrna Levels ◽  
Type Ii ◽  
Collagen Mrna ◽  
Type I Procollagen ◽  
Embryonic Chicken

Growth of embryonic chicken sternal chondrocytes in the presence of phorbol-12-myristate-13-acetate (PMA), a potent tumor promoter, resulted in a dramatic morphological change from spherical floating cells to adherent fibroblastic cells. This morphological change was accompanied by a quantitative switch from synthesis of cartilage-specific type II procollagen to type I procollagen. Type II procollagen mRNA levels decreased 10-fold in PMA-treated cells. Activation of type I collagen genes led to the accumulation of type I procollagen mRNA levels comparable to those of type II mRNA in these cells. However, only type I procollagen mRNA was translated. In addition to gene activation, unprocessed pro alpha 1(I) transcripts present at low levels in control chondrocytes were processed to mature mRNA species. Redifferentiation of PMA-treated chondrocytes was possible if cells were removed from PMA after the morphological change and cessation of type II procollagen synthesis but before detectable amounts of type I procollagen were synthesized. Production of type I collagen thus marks a late phase of chondrocyte "dedifferentiation" from which reversion is no longer possible. Redifferentiated cell populations contained 24-fold more pro alpha 1(II) collagen mRNA than pro alpha 1(I) collagen mRNA, but the rates of procollagen synthesis were comparable. This suggests that the PMA-mediated dedifferentiation of chondrocytes as well as their redifferentiation is under both transcriptional and posttranscriptional regulation.

scholarly journals Collagen expression in embryonic chicken chondrocytes treated with phorbol myristate acetate.

1985 ◽  
Vol 5 (6) ◽  
pp. 1415-1424 ◽  
Author(s):  
M H Finer ◽  
L C Gerstenfeld ◽  
D Young ◽  
P Doty ◽  
H Boedtker
Keyword(s):  
Morphological Change ◽  
Type I Collagen ◽  
Late Phase ◽  
Tumor Promoter ◽  
Type I ◽  
Mrna Levels ◽  
Type Ii ◽  
Collagen Mrna ◽  
Type I Procollagen ◽  
Embryonic Chicken

Growth of embryonic chicken sternal chondrocytes in the presence of phorbol-12-myristate-13-acetate (PMA), a potent tumor promoter, resulted in a dramatic morphological change from spherical floating cells to adherent fibroblastic cells. This morphological change was accompanied by a quantitative switch from synthesis of cartilage-specific type II procollagen to type I procollagen. Type II procollagen mRNA levels decreased 10-fold in PMA-treated cells. Activation of type I collagen genes led to the accumulation of type I procollagen mRNA levels comparable to those of type II mRNA in these cells. However, only type I procollagen mRNA was translated. In addition to gene activation, unprocessed pro alpha 1(I) transcripts present at low levels in control chondrocytes were processed to mature mRNA species. Redifferentiation of PMA-treated chondrocytes was possible if cells were removed from PMA after the morphological change and cessation of type II procollagen synthesis but before detectable amounts of type I procollagen were synthesized. Production of type I collagen thus marks a late phase of chondrocyte "dedifferentiation" from which reversion is no longer possible. Redifferentiated cell populations contained 24-fold more pro alpha 1(II) collagen mRNA than pro alpha 1(I) collagen mRNA, but the rates of procollagen synthesis were comparable. This suggests that the PMA-mediated dedifferentiation of chondrocytes as well as their redifferentiation is under both transcriptional and posttranscriptional regulation.

scholarly journals Regulation of type I collagen mRNA in lung fibroblasts by cystine availability

1998 ◽  
Vol 331 (2) ◽  
pp. 417-422 ◽  
Author(s):  
David C. RISHIKOF ◽  
Ping-Ping KUANG ◽  
Christine POLIKS ◽  
Ronald H. GOLDSTEIN
Keyword(s):  
Amino Acids ◽  
Steady State ◽  
Amino Acid ◽  
Type I Collagen ◽  
State Level ◽  
Lung Fibroblasts ◽  
Type I ◽  
Mrna Levels ◽  
Collagen Mrna ◽  
Amino Acid Availability

The steady-state level of α1(I) collagen mRNA is regulated by amino acid availability in human lung fibroblasts. Depletion of amino acids decreases α1(I) collagen mRNA levels and repletion of amino acids induces rapid re-expression of α1(I) mRNA. In these studies, we examined the requirements for individual amino acids on the regulation of α1(I) collagen mRNA. We found that re-expression of α1(I) collagen mRNA was critically dependent on cystine but not on other amino acids. However, the addition of cystine alone did not result in re-expression of α1(I) collagen mRNA. Following amino acid depletion, the addition of cystine with selective amino acids increased α1(I) collagen mRNA levels. The combination of glutamine and cystine increased α1(I) collagen mRNA levels 6.3-fold. Methionine or a branch-chain amino acid (leucine, isoleucine or valine) also acted in combination with cystine to increase α1(I) collagen mRNA expression, whereas other amino acids were not effective. The prolonged absence of cystine lowered steady-state levels of α1(I) collagen mRNA through a mechanism involving decreases in both the rate of gene transcription as assessed by nuclear run-on experiments and mRNA stability as assessed by half-life determination in the presence of actinomycin D. The effect of cystine was not mediated via alterations in the level of glutathione, the major redox buffer in cells, as determined by the addition of buthionine sulphoximine, an inhibitor of γ-glutamylcysteine synthetase. These data suggest that cystine directly affects the regulation of α1(I) collagen mRNA.

scholarly journals Immunohistochemical localization of procollagens. III. Type I procollagen antigenicity in osteoblasts and prebone (osteoid).

1981 ◽  
Vol 29 (7) ◽  
pp. 791-804 ◽  
Author(s):  
G M Wright ◽  
C P Leblond
Keyword(s):  
Bone Growth ◽  
Secretory Granules ◽  
Immunohistochemical Localization ◽  
Multivesicular Bodies ◽  
Type I ◽  
Frozen Sections ◽  
Type I Procollagen ◽  
The Matrix ◽  
Old Rats ◽  
Procollagen Iii

Frozen sections of unfixed tibia and mandibles from day-old rats were immunostained by the peroxidase-antiperoxidase technique after exposure to antisera against procollagen I or other collagenous materials. Light microscopic study of bone growth areas showed that procollagen I antigenicity was present in osteoblasts and prebone (osteoid), but not bone tissue; neither procollagen III nor collagen IV antigenicity were detected. The localization of procollagen I antigenicity within osteoblasts was then attempted in the electron microscope. Chopper slices of formaldehyde-fixed tibia from day-old rats were incubated with affinity-purified anti-procollagen I antibodies linked to peroxidase and were treated with hydrogen peroxide in the presence of 3,3'-diaminobenzidine. The dot-like reaction indicative of procollagen I antigenicity was found to be moderate in rough endoplasmic reticulum (rER) cisternae, strong in spherical and cylindrical Golgi distensions, and intense in prosecretory and secretory granules. Reactivity was also observed within the matrix of multivesicular bodies Thus an increasing gradient of procollagen I antigenicity occurs from rER cisternae through Golgi distensions to secretory granules, that is, along the presumed pathway of procollagen synthesis. The antigenicity present within rER cisternae is attributed to the precursor pro alpha (I) chains, while that in the cylindrical Golgi distensions, secretory granules, and prebone is attributed to procollagen itself. The antigenicity of multivesicular bodies suggests some degradation of pro alpha chains or procollagen.

Time-Varying Effect of Ascorbate on the Compressive Modulus and Gag Content of Chondrocyte-Seeded Agarose Cultures

1999 ◽  
Author(s):  
Robert L. Mauck ◽  
Michael A. Soltz ◽  
Beth Gilbert ◽  
Gerard A. Ateshian ◽  
Clark T. Hung ◽  
...  
Keyword(s):  
Mechanical Properties ◽  
Articular Chondrocytes ◽  
Biochemical Properties ◽  
Type I ◽  
Mrna Levels ◽  
Growth Plate Chondrocytes ◽  
Type I Procollagen ◽  
Cartilage Extracellular Matrix ◽  
The Matrix ◽  
Bovine Articular Chondrocytes

Abstract The generation of a cartilage substitute will require both the mechanical and biochemical properties of the substance to approach that of native tissue. Recent studies (Buschmann et al., 1992, 1995; Bader and Lee, 1997) have shown that agarose can provide a suitable environment for the production of a mechanically functional matrix. Over time cells seeded in this matrix alter the intrinsic mechanical properties of the matrix. Additionally, several studies have included such biochemical factors as vitamin C (ascorbate) to the culture medium. This biological effector has varied effects on chondrocyte matrix biosynthesis. Specifically, it has been shown to promote calcification in chick growth plate chondrocytes (Wu et al., 1989), while it increased both type II and type I procollagen mRNA in 5 day culture (Sandell and Daniel, 1988). Conversely, it has been shown in bovine culture that aggrecan mRNA levels rise steadily in monolayer culture without ascorbate. but rise quickly and then fall off in its presence (Hering et al., 1994). To assess the effect of ascorbate in our system, primary bovine articular chondrocytes seeded in agarose, we initiated a study to examine the effects of ascorbate on both the production of cartilage extracellular matrix and the development of mechanical properties over a 14 day culture period. Three groups were studied: agarose disks without cells (control) and cell-seeded agarose disks maintained in DMEM supplemented daily with and without 50 μg/ml of ascorbate. Glycosaminoglycan (GAG) and hydroxyproline contents of each disk were determined using standard colorimetric assays.

Abstract 7: Activation Of Neuronal At 1 R Exacerbates Hypertension-induced Reduction In Neuronal Function.

Hypertension ◽  
2020 ◽  
Vol 76 (Suppl_1) ◽  
Author(s):  
Jiaxi Xu ◽  
Eric D Lazartigues
Keyword(s):  
Cortical Neurons ◽  
Mrna Levels ◽  
Ang Ii ◽  
Neuronal Function ◽  
Salt Treatment ◽  
Bdnf Expression ◽  
Neurogenic Hypertension ◽  
Discs Large ◽  
Salt Hypertension ◽  
Cerebrovascular System

Hypertension has now been considered as one of the risk factors of Alzheimer's disease (AD), due to its contribution to the dysfunction of cerebrovascular system. To investigate its neuronal contribution, hypertension was induced in C57BL/6j male mice by either systemic infusion of Ang-II (600 ng/kg/min, s.c., 14 days) or DOCA-salt treatment (1 mg/g, s.c., 21 days), then markers for neuronal function were measured via qRT-PCR. In the hippocampus, Ang-II treatment significantly down-regulated the mRNA levels of BDNF (brain-derived neurotrophic factor) and DLG4 (discs large homolog 4, encoding PSD95), while DOCA-salt treatment only down-regulated BDNF expression (P<0.05 vs. sham, n=6). Notably, the expression of PI4KIIIβ, a key kinase for phosphatidylinositol-4,5-isphosphate (PIP 2 ) re-synthesis, was found to be markedly down-regulated in the hippocampus of both hypertension models (P<0.05 vs. sham, n=6). PI4K activity has been closely associated with the progression of neurodegenerative disorders, especially AD, therefore suggesting that reduction of neuronal function could be a part of the etiology of hypertension-related cognitive decline. We have demonstrated that neuronal AT 1 R plays pivotal role in the maintenance of neurogenic hypertension, and here we hypothesized that activation of AT 1 R could also exacerbate hypertension-induced reduction in neuronal function. In mice with DOCA-salt hypertension, the function of cortical neurons was shown to be improved by selective deletion of neuronal AT 1a R, as evidenced by significantly higher mRNA levels of BDNF and PI4KIIIβ, compared to the controls (P<0.05 vs. sham, n=6). To further study the possible involvement of neuronal AT 1 R in AD, 5хFAD mice were bred with mice with neuronal AT 1 R deletion (AT1NKO). AD-associated reduction of ACE2 protein, mainly in neurons, was found to be slightly ameliorated in the prefrontal cortex of 5хFAD-AT1NKO, compared to the age/sex-matched 5хFAD, showing by immunocytochemistry (24610 ±4182 vs. 13420 ±3720 AFU, n=6 slices). Although the detailed mechanism is still unknown, our data suggest that, neuron-expressing AT 1 R could participate in the development of hypertension-associated cognitive impairment and AD, independently of vascular AT 1 R.

Type I and Type III Collagen mRNA Levels in Kidney Regions of Old and Young Rats

Matrix ◽  
1993 ◽  
Vol 13 (4) ◽  
pp. 281-287 ◽  
Author(s):  
Itzhak Peleg ◽  
Ziv Greenfeld ◽  
Helena Cooperman ◽  
Shmuel Shoshan
Keyword(s):  
Type Iii Collagen ◽  
Type I ◽  
Mrna Levels ◽  
Type Iii ◽  
Collagen Mrna ◽  
Young Rats

scholarly journals Sustained hypoxia leads to the emergence of cells with enhanced growth, migratory, and promitogenic potentials within the distal pulmonary artery wall

2009 ◽  
Vol 297 (6) ◽  
pp. L1059-L1072 ◽  
Author(s):  
Maria G. Frid ◽  
Min Li ◽  
Meena Gnanasekharan ◽  
Danielle L. Burke ◽  
Miguel Fragoso ◽  
...  
Keyword(s):  
Pulmonary Artery ◽  
Mesenchymal Cell ◽  
Cell Types ◽  
Type I ◽  
Mrna Levels ◽  
Functional Characteristics ◽  
Cellular Mechanisms ◽  
Cell Markers ◽  
Type I Procollagen

All forms of chronic pulmonary hypertension (PH) are characterized by structural remodeling of the pulmonary artery (PA) media, a process previously attributed solely to changes in the phenotype of resident smooth muscle cells (SMC). However, recent experimental evidence in both systemic and pulmonary circulations suggests that other cell types, including circulating and local progenitors, contribute significantly to this process. The goal of this study was to determine if hypoxia-induced remodeling of distal PA (dPA) media involves the emergence of cells with phenotypic and functional characteristics distinct from those of resident dPA SMC and fibroblasts. In vivo, in contrast to the phenotypically uniform SMC composition of dPA media in control calves, the remodeled dPA media of neonatal calves with severe hypoxia-induced PH comprised cells exhibiting a distinct phenotype, including the expression of hematopoetic (CD45), leukocytic/monocytic (CD11b, CD14), progenitor (cKit), and motility-associated (S100A4) cell markers. Consistent with these in vivo observations, primary cell cultures isolated from dPA media of hypertensive calves yielded not only differentiated SMC, but also smaller, morphologically rhomboidal (thus termed here “R”) cells that transiently expressed CD11b, constitutively expressed the mesenchymal cell marker type I procollagen, expressed high mRNA levels of progenitor cell markers cKit, CD34, CD73, as well as for inflammatory mediators, IL-6 and MCP-1, and, with time in culture, gained expression of a myofibroblast marker, α-SM-actin. R cells exhibited highly augmented proliferative, migratory, invasive, and potent promitogenic capabilities, which were due, at least in part, to the production of PDGFs, SDF-1/CXCL12, and S100A4. These data suggest that the cellular mechanisms of dPA remodeling include the emergence of cells with phenotypic and functional characteristics markedly distinct from those of resident dPA cells.

scholarly journals Altered beta-actin gene expression in phorbol myristate acetate-treated chondrocytes and fibroblasts.

1985 ◽  
Vol 5 (6) ◽  
pp. 1425-1433 ◽  
Author(s):  
L C Gerstenfeld ◽  
M H Finer ◽  
H Boedtker
Keyword(s):  
Steady State ◽  
Type I Collagen ◽  
Type Ii Collagen ◽  
Tumor Promoter ◽  
Type I ◽  
Mrna Levels ◽  
Collagen Mrna ◽  
Beta Actin ◽  
Steady State Levels ◽  
Actin Mrna

Phorbol-12-myristate-13-acetate (PMA), a potent tumor promoter, was shown to have opposite effects on the cellular morphology and steady-state levels of beta-actin mRNA in embryonic chicken muscle fibroblasts and sternal chondrocytes. When fibroblasts were treated with PMA, they formed foci of densely packed cells, ceased to adhere to culture plates, and had significantly reduced levels of beta-actin mRNA and protein. Conversely, when treated with PMA, floating chondrocytes attached to culture dishes, spread out, and began to accumulate high levels of beta-actin mRNA and proteins. In the sternal chondrocytes the stimulation of the beta-actin mRNA production was accompanied by increased steady-state levels of fibronectin mRNAs and protein. These alterations were concomitant with a fivefold reduction in type II collagen mRNA and a cessation in its protein production. After fibronectin and actin mRNAs and proteins reached their maximal levels, type I collagen mRNA and protein synthesis were turned on. Removal of PMA resulted in reduced beta-actin mRNA levels in chondrocytes and in a further alteration in the cell morphology. These observed correlations between changes in cell adhesion and morphology and beta-actin expression suggest that the effect of PMA on cell shape and adhesion may result in changes in the microfilament organization of the cytoskeleton which ultimately lead to changes in the extracellular matrix produced by the cells.

scholarly journals Regulation of type I collagen mRNA levels in fibroblasts

1986 ◽  
Vol 157 (2) ◽  
pp. 433-439 ◽  
Author(s):  
Tilman VOSS ◽  
Paul BORNSTEIN
Keyword(s):  
Type I Collagen ◽  
Type I ◽  
Mrna Levels ◽  
Collagen Mrna
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