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Etude cytochimique des protides myéliniques

Cytochemical study of myelin proteids

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Summary

In the present study, it is shown that myelin and some perikaryal inclusions show a bluish fluorescence after glutaraldehyde fixation. This fluorescence can be extinguished by Sudan Black B and reinforced when calcium chloride is added to the fixation fluid; so, we think that this reaction must be due to the complex „lipoproteidic patterns“ found in sudanophil, PAS-positive inclusions, and in the myelin sheath.

In order to find out if similar results could be obtained, in vitro, after an identical treatment of different lipidic structures, at first, a chemical fractionation of the components of the rat brain was performed; then, after the isolation of a myelin fraction by ultracentrifugation, we identified its components by thin-layer chromatography. Controls were made directly on gangliosides, proteolipids and myelin proteins after extraction with suitable techniques.

We observe a marked reinforcement of the natural autofluorescence of lipidic components after glutaraldehyde treatment. This difference is clearly shown at the level of cerebrosides, phosphatides and proteolipids.

Electrophoresis and gel filtration before and after dialysis of the myelin extracts against organic solvents allow us to attribute the reaction mainly to the myelin proteids.

Résumé

Ce travail nous a permis de mettre en évidence une fluorescence bleuâtre de la myéline et de certaines inclusions péricaryales après fixation par le glutaraldehyde. L'observation de l'extinction de cette fluorescence par le noir Soudan B et de son renforcement par l'addition au fixateur de chlorure de calcium, nous a amené à penser que ce phénomène pourrait être imputé aux ≪édifices lipoprotidiques≫ complexes rencontrés dans les inclusions soudanophiles et PAS-positives, et dans la gaine de myéline.

En vue de vérifier si ce phénomène s'observait après traitement, in vitro, de diverses molécules lipidiques, nous avons fractionné chimiquement l'ensemble des constituants du cerveau du rat, puis isolé la myéline par ultracentrifugation et séparé ses composants par Chromatographie sur couche mince. Les résultats obtenus par ces deux types de méthodes ont été ensuite vérifiés par une analyse des gangliosides, des protéolipides et des protéines myéliniques après extraction selon des méthodes appropriées.

On observe un très net renforcement de l'autofluorescence naturelle des composés lipidiques grâce à l'action du glutaraldehyde. Cette différence est marquée au niveau des cérébrosides, des phosphatides et des protéolipides.

La technique d'électrophorèse et de filtration sur gel préalable ou après dialyse des extraits de la myéline contre des solvants organiques permet d'imputer ce phénomène aux protides myéliniques.

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  1. Laboratoire de Cytologie, Faculté des Sciences, Paris 5e et Centre de Microscopie Electronique, Hôpital de la Salpêtrière, Paris 13e

    Pierre L'Hermite

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L'Hermite, P. Etude cytochimique des protides myéliniques. Histochemie 22, 140–154 (1970). https://doi.org/10.1007/BF00303625

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