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

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.

2000 ◽  
Vol 85 (1) ◽  
pp. 124-133 ◽  
Author(s):  
Jennifer D. Wallace ◽  
Ross C. Cuneo ◽  
Per Arne Lundberg ◽  
Thord Rosén ◽  
Jens Otto Lunde Jørgensen ◽  
...  

To examine the interactions between acute exercise and GH on markers of bone and collagen turnover and to assess the potential for detecting GH abuse in athletes using these markers, we studied 17 aerobically trained males (age, 26.9 ± 1.5 yr). Sequential studies of exercise, GH administration, and GH withdrawal were undertaken. A randomized, controlled study of rest vs. exercise showed that exercise did not change serum osteocalcin; other markers of formation increased transiently (each P < 0.001): bone-specific alkaline phosphatase (+16.1%), carboxyterminal propeptide of type I procollagen (+14.1%), and procollagen III N-terminal extension peptide (+5.0%). The carboxyterminal cross-linked telopeptide of type I collagen, a bone resorption marker, increased 9.7% (P = 0.018) in response to exercise. A randomized, double blind, placebo-controlled, parallel study of recombinant human GH treatment (0.15 IU/kg·day) for 1 week increased serum osteocalcin (net increase preexercise, +10.0%; P = 0.017), carboxyterminal propeptide of type I procollagen (+17.6%; P = 0.002), procollagen III N-terminal extension peptide (+48.4%; P = 0.001), and carboxyterminal cross-linked telopeptide of type I collagen (53.3%; P = 0.009). Disappearance half-times after cessation of recombinant human GH for pre- and postexercise markers ranged from 248–770 h. We conclude 1) endurance exercise transiently activates bone and collagen turnover; 2) brief GH administration results in similar but quantitatively greater augmentation; and 3) these data will assist in designing a GH detection strategy.


1999 ◽  
Author(s):  
Robert L. Mauck ◽  
Michael A. Soltz ◽  
Beth Gilbert ◽  
Gerard A. Ateshian ◽  
Clark T. Hung ◽  
...  

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.


1968 ◽  
Vol 16 (5) ◽  
pp. 320-336 ◽  
Author(s):  
ERIC HOLTZMAN ◽  
REGINA DOMINITZ

The adrenalin-producing cells of the rat adrenal medulla have been studied by light and electron microscopy. Frozen sections of glutaraldehyde-perfused material were incubated for demonstration of "marker" enzymes for lysosomes (acid phosphatase, aryl sulfatase) and Golgi apparatus (thiamine pyrophosphatase). In addition, the uptake and fate of intravenously administered horseradish peroxidase was followed. Acid phosphatase activity is demonstrable in secretory granules, Golgi saccules, vesicles in the Golgi area and in the agranular tubules and cisternae (GERL) from which secretory granules appear to form at the inner surface of the Golgi apparatus. Endoplasmic reticulum with ribosomes on only one surface is closely apposed to both inner and outer aspects of the Golgi apparatus. Peroxidase is taken up in vesicles, tubules and "cup-like" bodies. The latter apparently transform into multivesicular bodies. A possible source of the acid phosphatase found in multivesicular bodies is the small vesicles from the Golgi apparatus or GERL.


1994 ◽  
Vol 267 (6) ◽  
pp. H2237-H2244 ◽  
Author(s):  
S. Besse ◽  
V. Robert ◽  
P. Assayag ◽  
C. Delcayre ◽  
B. Swynghedauw

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.


2021 ◽  
Vol 6 (2) ◽  
pp. 249-257
Author(s):  
A. Yu. Chumachenko ◽  
◽  
Е. G. Redka ◽  

According to modern data, the epiphyse (pineal gland) is an organ that combines the processes of adaptogenesis and immunogenesis, takes part in triggering stress reactions and determines the sequence of disorders in the body at different stages of stress development. Researchers consider the pineal gland to be the most important organizer of biological rhythms associated with photoperiodism and the organ that determines the stereotype of the organism. Its individual functions are rhythmically variable under the influence of the environment and age. The purpose of the study was to study the structural and functional changes of pineal cells of the pineal gland in rats at different stages of normal development, under the action of nitrates and the simultaneous action of nitrates and methylene blue. Materials and methods. In accordance with the purpose of the work, the study was carried out on 90 nonlinear white male rats of different ages. The animals were kept in the vivarium in equivalent conditions. Long-term exposure to nitrates on the body of animals was achieved by daily introduction into the drinking ration, starting from the 7th day of postnatal development of rats (after preliminary water purification), 120 mg/l sodium nitrate, that is, in a dose that is typical for many regions of Ukraine. When simulating the action of methylene blue, this substance was daily orally administered to the animals in doses: 0.1-0.15 ml of a 1% aqueous solution per 1 kg of body weight. Results and discussion. As a result of the 7-day action of nitrates in 14-day-old rats, structural changes were observed in the pineal gland, which corresponded to a decrease in the function of light cells and an increase in the functional activity of type II pinealocytes. The ultrastructure of the cytoplasm of type I pinealocytes contained poorly developed organelles and single secretory granules. In 45-day-old animals exposed to nitrates in light pinealocytes, pronounced disturbances in membrane organelles, primarily in mitochondria and the granular endoplasmic reticulum, were noted. The functional activity of dark pinealocytes increased during this period of the study. In the pineal parenchyma of 90-day-old rats after exposure to nitrates, the functional activity of type I pinealocytes was at a low level. The functional activity of dark pinealocytes was also weakened. Thus, as a result of the simultaneous action of nitrates and methylene blue in the pineal gland of 14-day-old rats, a tendency to gradual restoration of the structural and functional parameters of cells was observed. In 45-day-old animals, after the simultaneous action of nitrates and methylene blue, the ultrastructural data of pineal cells indicated numerous mitochondria and secretory granules in the cytoplasm. In the parenchyma of the pineal gland of 90-day-old rats after chronic action of nitrates and methylene blue at the ultrastructural level, no sharp changes in the cytoplasm and nucleus of light and dark pinealocytes were found in comparison with the control. Conclusion. The intake of nitrates in 14-day-old rats causes the development of a stress reaction, poorly developed organelles and signs of degranulation appear in the ultrastructure of light pinealocytes, however, the cytoplasm and nuclei of dark cells indicated an increase in function. In 45-day-old rats after exposure to nitrates, the signs of the stress reaction are enhanced. In the ultrastructure of the cytoplasm of light cells, pronounced violations of membrane organelles are determined. Enhanced function continues in dark pinealocytes. After the action of nitrates in 90-day-old rats, changes occur that are characteristic of the stage of depletion of the general adaptation syndrome, the result of which is a deep imbalance in the work of the pineal gland. The combined action of nitrates and methylene blue in 14-day-old animals helps to reduce the toxic effect and the strength of stress reactions in the pineal gland. In the ultrastructure of pinealocytes, the number of ribosomes, small secretory granules and mitochondria increases in comparison with the action of nitrates alone. In 45-day-old animals with the simultaneous intake of nitrates and methylene blue in the ultrastructure of melanotropic cells, the accumulation of secretory granules of the same size and electron density, an increase in the number of organelles and signs of restoration of the structure of the cytoplasm and nucleus are noted. The use of methylene blue against the background of long-term intake of nitrates in 90-day-old rats at the ultrastructural level of abrupt changes in the cytoplasm and nucleus of light and dark pinealocytes is not manifested in comparison with the control


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