DETECTION OF COMPLEX CARBOHYDRATES IN THE GOLGI APPARATUS OF RAT CELLS

A. Rambourg, W. Hernandez, C. P. Leblond

The Journal of Cell Biology · 1969 · 356 citations · 37 references

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TL;DR

Two electron‑microscopic methods were used to detect glycoproteins in rat cells: a sequential periodic acid–chromic acid–silver methenamine protocol and a brief chromic acid–phosphotungstic acid treatment. Both methods yielded identical results, showing a gradient of glycoprotein staining from the immature to the mature face of Golgi saccules, with the mature face and associated secretory granules strongly reactive, and indicating that Golgi saccules supply glycoproteins to secretion, lysosomal pathways, the cell coat, and basement membrane.

Abstract

Two methods used for the electron microscopic detection of glycoproteins were applied to a variety of cell types in the rat; one involved successive treatment of sections with periodic acid, chromic acid, and silver methenamine; and the other, a brief treatment with a chromic acid-phosphotungstic acid mixture. The results obtained with the two methods were identical and, whenever the comparison was possible, similar to those obtained with the periodic acid-Schiff technique of light microscopy. In secretory as well as in nonsecretory cells, parts of the Golgi apparatus are stained. The last saccule on one side of each Golgi stack is strongly reactive (mature face), and the last saccule on the other side shows little or no reactivity (immature face); a gradient of reactivity occurs in between these saccules. The more likely explanation of the increase in staining intensity is that carbohydrate is synthesized and accumulates in saccules as they migrate toward the mature face. In many secretory cells, the mature face is associated with strongly stained secretory granules. Other structures stained are: (1) small vesicles, dense and multivesicular bodies, at least some of which are presumed to be lysosomal in nature; (2) cell coat; and (3) basement membrane. The evidence suggests that the Golgi saccules provide glycoproteins not only for secretion, but also for the needs of the lysosomal system as well as for incorporation into the cell coat and perhaps basement membrane.

References

37