Experimental mucopolysaccharidosis: preservation and ultrastructural visualization of intralysosomal glycosaminoglycans by use of the cationic dyes cuprolinic blue and toluidine blue.
The cationic dyes Cuprolinic Blue (CB) and Toluidine Blue (TB) were used to preserve the intralysosomal storage material accumulating in tilorone-induced mucopolysaccharidosis. As shown in previous studies, the stored glycosaminoglycans (GAGs) are leached during the conventional fixation procedure, with the result that the lysosomes appear empty. In the present study, the liver, spleen, and cornea-conjunctiva of tilorone-treated rats were examined. The application of CB in the presence of 0.1 M or 0.3 M MgCl2 simultaneously with, or subsequently to the primary fixative yielded electron-dense precipitates within the storage lysosomes. When TB (0.1%) was added to the primary fixative, the storage lysosomes contained filamentous structures arranged in reticular patterns. With increasing TB concentrations (up to 1%) the lysosomes increasingly often showed apparently amorphous storage material which was continuous with the reticular filamentous structures. Similar ultrastructural patterns were obtained with GAG-TB complexes prepared in vitro. The intralysosomal storage material preserved by TB is interpreted as GAG-TB precipitates. In conclusion, the use of CB provides a method which allows direct cytochemical demonstration of the subcellular sites of GAG-storage. The use of TB represents an easy method to obtain electron micrographs pathognomonic of the mucopolysaccharidosis induced by tilorone and congeners. Either method may be helpful to detect this adverse drug effect at the subcellular level.
Lüllmann-Rauch
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Department of Anatomy, University of Kiel, Federal Republic of Germany.
D000818:Animals; D004396:Coloring Agents; D003228:Conjunctiva / Q000032:analysis / Q000648:ultrastructure; D003315:Cornea / Q000032:analysis / Q000648:ultrastructure; D005260:Female; D005404:Fixatives; D006025:Glycosaminoglycans / Q000032:analysis*; D007211:Indoles; D008099:Liver / Q000032:analysis / Q000648:ultrastructure; D008247:Lysosomes / Q000032:analysis* / Q000648:ultrastructure; D009083:Mucopolysaccharidoses / Q000139:chemically induced / Q000473:pathology*; D009942:Organometallic Compounds; D051381:Rats; D011919:Rats, Inbred Strains; D013154:Spleen / Q000032:analysis / Q000648:ultrastructure; D013994:Tilorone; D014048:Tolonium Chloride
D004396:Coloring Agents; D005404:Fixatives; D006025:Glycosaminoglycans; D007211:Indoles; D009942:Organometallic Compounds; D014048:Tolonium Chloride; C015445:copper phthalocyanine; D013994:Tilorone