Abstract
In normal dogs and in dogs treated with 20 or 30 U.S.P. parathyroid extract for 5 and 3–4 days, respectively, the glycosaminoglycans of compact bone tissue were identified using the cetylpyridinium chloride precipitation method, and the concentrations of total hexosamines and the hexosamines corresponding to cetylpyridinium chloride precipitable acid glycosaminoglycans were determined. Further, the glycosaminoglycan pattern of the epiphyseal plate and the incorporation of35S-sulphate into the glycosaminoglycans of bone tissue and epiphyseal cartilage after administration of35S-sulphatein vivo was studied.
In compact bone tissue, the hexosamines corresponding to acid glycosaminoglycans constuted approximately one third of the total hexosamine concentration and approximately 0.05–0.06% of the total dry weight. The main component of the acid glycosaminoglycans in bone was chondroitin-4-sulphate. This was sulphated to a higher degree and also of a higher molecular weight than thechondroitin sulphate of the epiphyseal cartilage, which in accordance with earlier investigations was found to have infrared characteristics of both chondroitin-4-sulphate and chondroitin-6-sulphate, with the former dominating. The molecular weights of the main part of bone chondroitin sulphate ranged from approximately 45,000 to 56,000. A small component of the bone glycosaminoglycans was hyaluronic acid.
Large regularly recurring differences in the specific activity of fractions with differences in molecular weight in the condroitin sulphate of bone tissue and epiphyseal cartilage were noted.
Treatment of the dogs with parathyroid extract gave no effect on the molecular weights of the chondroitin sulphate of the bone matrix or of the epiphyseal cartilage. Nor was there any unequivocal effect on the concentrations of total hexosamines or on the acid glycosaminoglycans. No evident stimulatory or depressant effect on the incorporation of35S-sulphate into the chondroitin sulphate or in the molecular distribution of newly sulphated and/or synthesized molecules of the condroitin sulphate within these tissues occurd.
Résumé
Les glycosaminoglycanes de l'os compact ont été identifiés, par la méthode de précipitation au chlorure de cetylpyridinium, chez le chien normal et des chiens soumis à 20 ou 30 U.S.P. d'extrait parathyroïdien pendant 5 et 3–4 jours. Les concentrations d'héxosamines totales ainsi que les héxosamines, en rapport avec les glycoaminoglycanes acides, précipités par chlorure de cetylpyridinium, sont déterminées. En outre, la répartition du glycosaminoglycane de la zone d'ossification épiphysaire ainsi que l'incorporation du35S-sulfate dans les glycosaminoglycanes du tissu osseux et du cartilage épiphysaire, après administration de35S-sulfatein vivo, ont été étudiées.
Dans l'os compact, les héxosamines, en rapport avec les glycosaminoglycances acides, constitutent environ un tiers de la concentration totale en héxosamine et environ 0,05–0,06% du poids sec total. Le constituant principal des glycosaminoglycances acides osseux est formé par le chondroitine-4-sulfate. Ce dernier est plus riche en sulfate et présente un poids moléculaire plus élevé que le chondroitine sulfate du cartilage épiphysaire, qui, selon des travaux antérieurs, présente des caractéristiques infra-rouges du chondroitine-4-sulfate et du chondroitine-6-sulfate, avec prédominance du premier. Les poids moléculaires du chondroitine sulfate osseux varient surtout d'environ 45000 et 56000. L'acide hyaluronique constitute une faible fraction des glycosaminoglycanes osseux.
Des différences marquées de l'activité spécifique des fractions de cohondroitine sulfate de l'os et du cartilage épiphysaire, à poids moléculaires variables, on tété notées de façon répétée. L'administration d'extrait parathyroïdien à des chiens n'a pas d'effet sur les poids moléculaires en chondroitine sulfate de l'os ou du cartilage épiphysaire. Elle n'influence pas non plus les concentrations en héxosamines totales ou en glycosaminoglycances acides. Dans ces tissus, il ne se produit pas d'effect de stimulation ou de dépression concernant l'incorporation de35S-sulfate dans le chondroitine sulfate our sur la séparation moléculaire de molécules transformées en sulfates et/ou de molécules synthétiques de sulfate chondroitine.
Zusammenfassung
An normale Hunden und an Hunden die unter 5 respektive 3–4 Tagen 20 oder 30 U.S.P./kg Parathyreoidea-Extract erhalten hatten, wurden im kompakten Knochen die Glykosaminoglykane unter Anwendung der Cetylpyridiniumchlorid-Fällungsmethode identifiziert und die Konzentration des Total-Hexosamins und des Hexosamins, entsprechend den Cetylpyridiniumchlorid fällbaren sauren Glykosaminoglykanen, wurde bestimmt. Außerdem wurden das Glykosaminoglykanmuster der Epiphysenplatte und der Einbau von35S-Sulfat in die Glykosaminoglykane des Knochengewebes und des Epiphysenknorpels nach Zufuhr von35S-Sulfatin vivo studiert.
Im kompakten Knochengewebe macht des Hexosamin, entsprechend den sauren Glykosaminoglykanen, ungefähr ein Drittel der totalen Hexosaminkonzentration und ungefähr 0,05–0,06% des totalen Trockengewichtes aus. Der Hauptanteil der sauren Glykosaminoglykane im Knochen war Chondroitin-4-Sulfat. Dieses war in höherem Grad sulfatiert und hatte ein höheres Molekülgewicht als das Chondroitinsulfat der Epiphysenplatte, welches, in Übereinstimmung mit früheren Untersuchungen, Infrarot spektra kennzeichnend für sowohl Chondroitin-4-sulfat als auch für Chondroitin-6-sulfat, das Erstere überwiegend, hatte. Das Molekülgewicht des Hauptanteiles des Knochen-Chondroitinsulfates lag zwischen ungefähr 45000–56000. Ein kleiner Teil der Knochen-Glykosaminoglykane war Hyaluronsäure.
Sowohl im Knochengewebe als auch im Epiphysenknorpel wurde in verschiedenen Fraktionen des Chondroitinsulfates, mit unterschiedlichem Molekülgewicht, grße und regelmäßig reproduzierbare Unterschiede in der spezifischen Aktivität gefunden.
Behandlung der Hunde mit Parathyreoideaextrakt gab keinen Ausschlag in den Molekülgewichten des Chondroitinsulfates, weder des Knochens noch des Epiphysenknorpels. Ebenso wurde kein eindeutiger Effekt auf die Konzentration des totalen Hexosamins oder der sauren Glykosaminoglykane gefunden. Kein offenbarer, weder anregender noch senkender, Effekt auf den Einbau von35S-Sulfat in das Chondroitinsulfat oder in der molekularen Verteilung der neulich sulfatierten und/oder synthetisierten Moleküle des Chondroitinsulfates dieser Gewebe wurde gefunden.
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Abbreviations
- CPC:
-
cetylpyridinium chloride
- CP:
-
cetylpyridinium
- EDTA:
-
disodium ethylenediaminetetra-acetic acid
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The nomenclature ofBalasz andJeanloz (1965) has been followed.
This work was reported, in part, at the meeting of Svenska Patologföreningen in Stockholm, Sweden, December 26th–27th, 1965; see Nordisk Medicin 75, 434–435 (1965); and at the Fourth European Symposium on Calcified Tissues, Leiden, Noordwijk aan Zee, The Netherlands, March 28th–April 1st. 1966; see Exerpta Medica Foundation, International Congress Series No 120, 1966, p. 55–56.
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Hjertquist, SO., Verjlens, L. The glycosaminoglycans of dog compact bone and epiphyseal cartilage in the normal state and in experimental hyperparathyroidism. Calc. Tis Res. 2, 314–333 (1968). https://doi.org/10.1007/BF02279220
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DOI: https://doi.org/10.1007/BF02279220