scholarly journals Studies on the incorporation of [U-14C]glucose and [35S]sulphate into the acid glycosaminoglycans of neonatal rat skin

1970 ◽  
Vol 119 (5) ◽  
pp. 885-893 ◽  
Author(s):  
T. E. Hardingham ◽  
C. F. Phelps
Keyword(s):  
Uronic Acid ◽  
Heparan Sulphate ◽  
Neonatal Rat ◽  
Rat Skin ◽  
Acid Biosynthesis ◽  
Keratan Sulphate ◽  
Sulphated Glycosaminoglycans ◽  
Acid Glycosaminoglycan ◽  
Acid Glycosaminoglycans

1. The incorporation of [35S]sulphate in vivo into the acid-soluble intermediates extracted from young rat skin showed three sulphated hexosamine-containing components. 2. The rates of synthesis of these components were determined in vivo by measuring the incorporation of radioactivity from [U-14C]glucose into their isolated hexosamine moieties. 3. The incorporation of radioactivity from [U-14C]glucose into the isolated hexosamine and uronic acid moieties of the acid glycosaminoglycans was also measured. These results, combined with those obtained on the intermediary pathways of hexosamine and uronic acid biosynthesis previously determined in this tissue, indicated that the acid-soluble sulphated hexosamine-containing components were not precursors of the sulphated hexosamine found in the acid glycosaminoglycans. 4. The rates of synthesis of the acid glycosaminoglycan fractions were calculated from the incorporation of radioactivity from [U-14C]glucose into the hexosamine moiety. The sulphated components containing principally dermatan sulphate, chondroitin 6-sulphate and in smaller amounts, chondroitin 4-sulphate, heparan sulphate and heparin appeared to be turning over about twice as rapidly as hyaluronic acid and about four times as rapidly as the small keratan sulphate fraction. The relative rates of synthesis of the sulphated glycosaminoglycans were calculated from the incorporation of [35S]sulphate and were in agreement with those from 14C-labelling studies.

scholarly journals In Vivo Metabolism of Topically Applied Benzo[a]pyrene-4,5-oxide in Neonatal Rat Skin

1984 ◽  
Vol 82 (4) ◽  
pp. 378-380 ◽  
Author(s):  
Benjamin J del Tito ◽  
Hasan Mukhtar ◽  
David R Bickers
Keyword(s):  
Neonatal Rat ◽  
In Vivo Metabolism ◽  
Rat Skin

scholarly journals Detection of secondary structure in glycosaminoglycans via the1H n.m.r. signal of the acetamido NH group

1982 ◽  
Vol 207 (1) ◽  
pp. 139-144 ◽  
Author(s):  
J E Scott ◽  
F Heatley
Keyword(s):  
Secondary Structure ◽  
Chondroitin Sulphate ◽  
Heparan Sulphate ◽  
Periodate Oxidation ◽  
Close Approach ◽  
Dimethyl Sulphoxide ◽  
Space Filling ◽  
Keratan Sulphate ◽  
Inorganic Cations ◽  
Acid Glycosaminoglycans

Two simple methods for dissolving salts of acid glycosaminoglycans with inorganic cations (e.g. Li+ and Na+) in dry dimethyl sulphoxide are described. Complete n.m.r. spectra of, e.g., Na+ and Li+ salts of chondroitin sulphate and keratan sulphate were obtained on these solutions. In [2H6]dimethyl sulphoxide the NH resonance of 2-acetamido-2-deoxy hexosides is in the range 7.2-8.0 delta, but is downfield (8.3-9.3 delta) when the NH is H-bonded to -CO2-. Heparan sulphate shows two NH resonances, of which one (at 8.3 delta) is probably indicative of H-bonding. Space-filling models show that a very close approach of NH to -CO2- across the alpha-glucosaminidic bond is possible, and a solution configuration for heparan sulphate is proposed. The n.m.r. results are entirely compatible with interpretations of periodate-oxidation kinetics, based on H-bonded secondary structures present in hyaluronate and chondroitin sulphates, but not in dermatan (or keratan) sulphate.

scholarly journals Molecular distinctions between heparan sulphate and heparin. Analysis of sulphation patterns indicates that heparan sulphate and heparin are separate families of N-sulphated polysaccharides

1985 ◽  
Vol 230 (3) ◽  
pp. 665-674 ◽  
Author(s):  
J T Gallagher ◽  
A Walker
Keyword(s):  
Uronic Acid ◽  
Heparan Sulphate ◽  
Analytical Data ◽  
Gel Filtration ◽  
Published Data ◽  
Sulphated Polysaccharides ◽  
Close Proximity ◽  
Sulphated Glycosaminoglycans ◽  
Very High ◽  
High Voltage Electrophoresis

Heparan sulphate and heparin are chemically related alpha β-linked glycosaminoglycans composed of alternating sequences of glucosamine and uronic acid. The amino sugars may be N-acetylated or N-sulphated, and the latter substituent is unique to these two polysaccharides. Although there is general agreement that heparan sulphate is usually less sulphated than heparin, reproducible differences in their molecular structure have been difficult to identify. We suggest that this is because most of the analytical data have been obtained with degraded materials that are not necessarily representative of complete polysaccharide chains. In the present study intact heparan sulphates, labelled biosynthetically with [3H]glucosamine and Na2(35)SO4, were isolated from the surface membranes of several types of cells in culture. The polysaccharide structure was analysed by complete HNO2 hydrolysis followed by fractionation of the products by gel filtration and high-voltage electrophoresis. Results showed that in all heparan sulphates there were approximately equal numbers of N-sulpho and N-acetyl substituents, arranged in a similar, predominantly segregated, manner along the polysaccharide chain. O-Sulphate groups were in close proximity to the N-sulphate groups but, unlike the latter, the number of O-sulphate groups could vary considerably in heparan sulphates of different cellular origins ranging from 20 to 75 O-sulphate groups per 100 disaccharide units. Inspection of the published data on heparin showed that the N-sulphate frequency was very high (greater than 80% of the glucosamine residues are N-sulphated) and the concentration of O-sulphate groups exceeded that of the N-sulphate groups. We conclude from these and other observations that heparan sulphate and heparin are separate families of N-sulphated glycosaminoglycans.

scholarly journals Absence of keratan sulphate from skeletal tissues of mouse and rat

1985 ◽  
Vol 228 (2) ◽  
pp. 443-450 ◽  
Author(s):  
G Venn ◽  
R M Mason
Keyword(s):  
Monoclonal Antibody ◽  
Degradation Products ◽  
Chondroitin Sulphate ◽  
Lumbar Disc ◽  
Heparan Sulphate ◽  
Costal Cartilage ◽  
Animal Experiments ◽  
Keratan Sulphate

The absence of keratan sulphate synthesis from skeletal tissues of young and mature mice and rats has been confirmed by (1) analysis of specific enzyme degradation products of newly synthesized glycosaminoglycans, and (2) immunohistochemistry and radioimmunoassay using a monoclonal antibody directed against keratan sulphate. Approx. 98% of the [35S]glycosaminoglycans synthesized in vivo by mouse and rat costal cartilage, and all of those of lumbar disc, are chondroitin sulphate. The remainder in costal cartilage were identified as heparan sulphate in mature rats. In contrast, [35S]glycosaminoglycans synthesized by cornea of both species comprised both chondroitin sulphate and keratan sulphate. In mice keratan sulphate accounted for 12-25% and in rats 40-50% of the total [35S]glycosaminoglycans, depending on the age of the animal. Experiments in vitro with organ culture of cartilage and cornea confirm these results. Absence of keratan sulphate from mouse costal cartilage and lumbar disc D1-proteoglycans was corroborated by inhibition radioimmunoassay with the monoclonal antibody MZ15 and by lack of staining for keratan sulphate in indirect immunofluorescence studies using the same antibody.

scholarly journals The glycosaminoglycans of neonatal rat skin

1970 ◽  
Vol 117 (5) ◽  
pp. 813-818 ◽  
Author(s):  
T. E. Hardingham ◽  
C. F. Phelps
Keyword(s):  
Post Partum ◽  
Cetylpyridinium Chloride ◽  
Heparan Sulphate ◽  
Neonatal Rat ◽  
Dermatan Sulphate ◽  
Short Chain Length ◽  
Low Degree ◽  
Cellulose Column ◽  
Testicular Hyaluronidase ◽  
Acid Glycosaminoglycans

The acid glycosaminoglycans were extracted from the skins of young rats less than 1 day post partum. The isolated products were fractionated by a cetylpyridinium chloride–cellulose column technique and identified by chemical analysis, electrophoretic mobility and susceptibility to testicular hyaluronidase digestion. Hyaluronic acid (56%) dermatan sulphate (15.6%) and chondroitin 6-sulphate (9.1%) were the major components, but chondroitin 4-sulphate, heparan sulphate and heparin were also present, together with two further fractions tentatively suggested to be a heparan sulphate-like fraction and a dermatan sulphate fraction, both of short chain length or low degree of sulphation.

scholarly journals 1H nuclear-magnetic-resonance spectra of the methyl group of the acetamido moiety and the structure of acid glycosaminoglycans in solution

1979 ◽  
Vol 181 (2) ◽  
pp. 445-449 ◽  
Author(s):  
J E Scott ◽  
F Heatley
Keyword(s):  
Nuclear Magnetic Resonance ◽  
Heparan Sulphate ◽  
Periodate Oxidation ◽  
Evidence Based ◽  
Keratan Sulphate ◽  
Methyl Group ◽  
Nuclear Magnetic Resonance Spectra ◽  
1H Nuclear Magnetic Resonance ◽  
Acid Glycosaminoglycans ◽  
Chemical Evidence

The 1H resonances of the methyl group in the acetamido moiety of several types of glycosaminoglycans are reported at 300 MHz in 2H2O. Dermatan sulphates with various L-iduronate/D-glucuronate ratios are compared with chrondroitin sulphates with various contents and positions of substitution of sulphate esters. Hyaluronate oligomers are compared with 2-acetamido-2-deoxy-D-glucose, and with heparan sulphate and keratan sulphate. The major determinant of the chemical shift of the acetamido methyl resonance is the closeness of approach between carboxy groups and the acetamido group, in agreement with chemical evidence based on periodate-oxidation kinetics.

scholarly journals The synthesis of glycosaminoglycans by cultures of rabbit corneal endothelial and stromal cells

1976 ◽  
Vol 158 (3) ◽  
pp. 567-573 ◽  
Author(s):  
B Y J T Yue ◽  
J L Baum ◽  
J E Silbert
Keyword(s):  
Hyaluronic Acid ◽  
Stromal Cells ◽  
Heparan Sulphate ◽  
Deae Cellulose ◽  
Dermatan Sulphate ◽  
Keratan Sulphate ◽  
Monolayer Cultures ◽  
Sulphated Glycosaminoglycans ◽  
Cellulose Column ◽  
Sulphated Glycosaminoglycan

Confluent monolayer cultures of rabbit corneal endothelial and stromal cells were incubated independently with [35S]sulphate and [3H]glucosamine for 3 days. AFter incubation, labelled glycosaminoglycans were isolated from the growth medium and from a cellular fraction. These glycosaminoglycans were further characterized by DEAE-cellulose column chromatography and by sequential treatment with various glycosamino-glycan-degrading enzymes. Both endothelial and stromal cultures synthesized hyaluronic acid as the principal product. The cell fraction from the stromal cultures, however, had significantly less hyaluronic acid than that from the endothelial cultures. In addition, both types of cells synthesized a variety of sulphated glycosaminoglycans. The relative amounts of each sulphated glycosaminoglycan in the two cell lines were similar, with chondroitin 4-sulphate, chondroitin 6-sulphate and dermatan sulphate as the major components. Heparan sulphate was present in smaller amounts. Keratan sulphate was also identified, but only in very small amounts (1-3%). The presence of dermatan sulphate and the high content of hyaluronic acid are similar to the pattern of glycosaminoglycans seen in regenerating or developing tissues, including cornea.

scholarly journals Human liver N-acetylglucosamine-6-sulphate sulphatase. Catalytic properties

1987 ◽  
Vol 246 (2) ◽  
pp. 355-365 ◽  
Author(s):  
C Freeman ◽  
J J Hopwood
Keyword(s):  
Enzyme Activity ◽  
Carboxy Group ◽  
Human Liver ◽  
Catalytic Mechanism ◽  
Heparan Sulphate ◽  
Catalytic Efficiency ◽  
Keratan Sulphate ◽  
Ph Response ◽  
A Minor

Kinetic parameters (Km and kcat.) of the two major forms (A and B) and a minor form (C) of human liver N-acetylglucosamine-6-sulphate sulphatase [Freeman, Clements & Hopwood (1987) Biochem. J. 246, 347-354] were determined with a variety of substrates matching structural aspects of the physiological substrates in vivo, namely heparin, heparan sulphate and keratan sulphate. Enzyme activity is highly specific towards glucosamine 6-sulphate or glucose 6-sulphate residues. More structurally complex substrates, in which several aspects of the aglycone structure of the natural substrate were maintained, are hydrolysed with catalytic efficiencies up to 3900 times above that observed for the monosaccharide substrate N-acetylglucosamine 6-sulphate. Forms A and B both desulphate substrates derived from keratan sulphate and heparin. Aglycone structures that influence substrate binding and/or enzyme activity were penultimate-residue 6-carboxy and 2-sulphate ester groups for heparin-derived substrates and penultimate-residue 6-sulphate ester groups for keratan sulphate-derived substrates. The 4-hydroxy group of the N-acetylglucosamine 6-sulphate or the 2-sulphaminoglucosamine 6-sulphate under enzymic attack is involved in the catalytic mechanism. The presence of a 2-amino group in place of a 2-acetamido or a 2-sulphoamino group considerably decreases the catalytic efficiency of the sulphatase, particularly in the absence of a penultimate-aglycone-residue 6-carboxy group. Both forms A and B are exo-enzymes, since activity towards internal sulphate ester bonds was not observed. The effect of incubation pH on enzyme activity towards the variety of substrates evaluated was complex and dependent on substrate aglycone structure. The presence of aglycone 2-sulphate ester, 6-carboxy group and 6-sulphate ester groups on the glucosamine 6-sulphate residue under attack considerably affects the pH response. Sulphate and phosphate ions are potent inhibitors of enzyme activity.

scholarly journals Heterogeneous distribution of antithrombin-binding sites in rat brain heparan sulphate proteoglycans

1991 ◽  
Vol 280 (2) ◽  
pp. 393-397 ◽  
Author(s):  
A A Horner
Keyword(s):  
Binding Sites ◽  
Heparan Sulphate ◽  
Deae Cellulose ◽  
Reciprocal Relationship ◽  
Rat Skin ◽  
Heterogeneous Distribution ◽  
Site Distribution ◽  
Heparan Sulphate Proteoglycans ◽  
High Binding Affinity

Heparan sulphates with high binding affinity for antithrombin (HA-HS), labelled in vivo with [35S]sulphate, were extracted from rat brains and purified by chromatography on DEAE-cellulose and on antithrombin-agarose. HA-HS proteoglycans (HA-HSPG) were then separated from HA-HS chains on Sepharose CL-6B. The total HA-HSPG product was rechromatographed on antithrombin-agarose. Six HA-HSPG subfractions with differing degrees of affinity for antithrombin were recovered and treated with NaOH to release their chains. Rechromatography of these six 35S-labelled HS chain preparations on antithrombin-agarose showed that their proportions of chains with no affinity for antithrombin (NA-HS chains) ranged from 36 to 71%. There was a reciprocal relationship between the proportion of NA-HS chains in each HA-HSPG subfraction and the degree of affinity for antithrombin of the rest of its chains (assessed relative to 3H-labelled HA-heparin chains with which they were co-chromatographed). Similar characteristics of antithrombin-binding-site distribution apply to HA-heparin proteoglycans from rat skin studied previously [Horner (1987) Biochem. J. 244, 693-698]. The data suggest that the sites at which 3-O-sulphation of some glucosamine N-sulphate residues occurs in the Golgi complex of brain cells (probably endothelial cells) which synthesize HA-HSPGs (as in mast cells, which synthesize HA-heparin PGs) are distributed sparsely but not randomly.

In vivo diffusion tensor imaging of the neonatal rat brain development

2013 ◽  
Vol 44 (S 01) ◽  
Author(s):  
M Breu ◽  
D Reisinger ◽  
D Wu ◽  
Y Zhang ◽  
A Fatemi ◽  
...  
Keyword(s):  
Diffusion Tensor Imaging ◽  
Brain Development ◽  
Rat Brain ◽  
Diffusion Tensor ◽  
Neonatal Rat ◽  
Neonatal Rat Brain
Sign in / Sign up

Export Citation Format

Share Document