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Studies on matrix proteins of normal and lathyritic rat bone and dentine

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Abstract

The collagen in bone and soft tissues of lathyritic animals is known to be more soluble than that of normal animals when traditional methods of extraction are employed. To ascertain whether a similar phenomenon occures in lathyritic dentine collagen, the solubility of dentine matrix collagen was investigated in two solvents. Young adult male rats were maintained on a lathyrogenic diet for 50 days, during which time the major part of the dentine matrix collagen of the incisors was shown to have turned over completely. Decalcified dentine matrix collagen was prepared by exhaustively dialysing cleaned incisors against 0.5M acetic acid, and the non-dialysable matrix was further extracted with the same solvent, and then with 5 M guanidine hydrochloride. The dentine matrix collagen of both normal and lathyritic animals proved to be insoluble in 0.5M acetic acid; guanidine hydrochloride extraction solubilized approximately 20% of the lathyritic matrix collagen as well as a quantity of non-collagen protein, in contrast to the soft tissue and bone collagens of lathyritic animals in which greater amounts of collagen may be solubilized by this procedure. Fractionation of the guanidine hydrochloride extract from the lathyritic animals by carboxymethyl cellulose chromatography revealed an unexpectedly high ratio of α1 to α2 components considerably greater than the 2∶1 ratio demonstrated for lathyritic rat bone. These data provide evidence for significant differences between bone and dentine collagens.

Résumé

Le collagène de l'os et des tissus mous des animaux lathyrique est plus soluble que celui d'animaux normaux, en utilisant des méthodes d'extraction classiques. Pour vérifier s'il existe und phénomène similaire au niveau du collagène dentinaire lathyrique, la solubilitè due collagène matricielle dentinaire est étudiée dans deux solvants. De jeunes rats mâles adultes sont soumis à un régime lathyrogénique pendant 50 jours, période pendant laquelle la majeure partie du collagène dentinaure des incisives semble être formée. Le collagène dentinaire décalcifié est préparé par dialyse d'incisives nettoyées avec de l'acide acétique 0.5M. et la matrice non-dialysable est extraite par le même solvant, suivi d'hydrochlorure de guanidine 5M. Le collagène dentinaure des animaux et lathyriques est insoluble dans l'acide acétique 0.5M: l'extraction par l'hydrochlorure de guanidine solubilise environ 20% du collagène lathyrique, ainsi qu'une certaire quantité de protéine non-callagénique, contrairement au collagène de l'os et des tissus mous d'animaux lathyriques, où de plus grandes quantités de collagène peuvent être solubilisées par ce procédé. Le fractionnement de l'extrait par hydrochlorure de guanidine des animaux lathyriques à l'aide de la chromatographie à cellulose de carboxyméthyle donne un rapport anormalement élevé des composés α1 et α2, qui est nettement plus grand que le rapport 2∶1 de l'os de rat Lythyrique. Ces résultats montrent des différences nettes entre les collagènes osseux et dentinaire.

Zusammenfassung

Es ist bekannt, daß das Collagen in Knochen und Weichgeweben von lathyritischen Tieren besser löslich ist als dasjenige normaler Tiere, wenn traditionalle Extraktionsmethoden verwendet werden. Um festzustellen, ob sich das lathyritische Dentincollagen ähnlich verhalte, wurde die Löslichkeit des Collagens der Dentinmatrix in zwei Lösungsmitteln untersucht. Junge erwachsene männliche Ratten erhielten während 50 Tagen eine lathyrogene Nahrung; es konnte nachgewiesen werden, daß während dieser Zeit das meiste Collagen der Dentin-Matrix der Schneidezähne völlig umgebaut wurde. Calciumfreies Dentinmatrix-Collagen wurde erhalten, indem gereinigte Schneidezähne ausgiebig mit 0,5 M Essigsäure dialysiert wurden; die nicht-dialysierbare Matrix wurde weiter mit demselben Lösungsmittel und dann mit 5M Guanidinhydrochlorid extrahiert. Das Collagen der Dentinmatrix von normalen und lathyritischen Tieren war in 0,5 M Essigsäure unlöslich; bei der Extraktion mit Guanidinhydrochlorid lösten sich etwa 20% des lathyritischen Matrix-Collagens sowie ein Teil nicht-collagenen Proteins, im Gegensatz zum Collagen der Weichgewebe und Knochen von lathyritischen Tieren, bei welchen mit diesem Verfahren größere Mengen von Collagen gelöst werden können. Die Fraktionierung des guanidinhydrochlorid-Extraktes von lathyritischen Tieren mittels Carboxymehtyl-Cellulose-Chromatographie zeigte ein unerwartet hohes Verhältnis der α1 und α2 zueinander; es war beträchtlich höher als das 2∶1-Verhältnis, das bei lathyritischen Rattenknochen nachgewiesen wurde. Diese Befunde zeigen signifikante Unterschiede zwischen Knochen- und Dentincollagen auf.

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Authors and Affiliations

  1. Department of Chemistry, University of Wisconsin, Eau Claire

    David J. Carmichael & Carole M. Dodd

  2. Faculty of Dentistry, University of Alberta, Edmonton, Alberta, Canada

    Charles F. Nawrot

Authors
  1. David J. Carmichael
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  2. Carole M. Dodd
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  3. Charles F. Nawrot
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Carmichael, D.J., Dodd, C.M. & Nawrot, C.F. Studies on matrix proteins of normal and lathyritic rat bone and dentine. Calc. Tis Res. 14, 177–194 (1974). https://doi.org/10.1007/BF02060294

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