Detection of Complex Carbohydrates in the Golgi Apparatus of Rat Cells

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.

Cytochemical Studies of Ameloblasts and the Surface Layer of Enamel of the Rat Incisor at the Maturation Stage

“…4, 13). The Cr-PTA reaction of Golgi cisterns was stronger at the inner cisterns that corresponded to the mature face of the Golgi apparatus (RAMBOURG, 1969) and many small vesicles associating with the inner c isterns were stained with Cr-PTA in different intensity (Fig. 13).…”

“…PEASE (1966PEASE ( , 1970, RAMBOURG, HERNANDEZ andLEBLOND (1969), andRAMBOURG (1969) presented a view that PTA at low pH is reasonably specific for carbohydrates, suggesting that the staining reaction is due to the presence of glycoproteins containing sialic acid residues.…”

“…Cr-PTA and PA-silver are thought essentially to stain the structures including the same substances though PA-silver nonspecifically stains other structures such as nuclei, nucleoli and ribosomes as well (RAMBOURG, HERNANDEZ and LEBLOND, 1969;RAMBOURG, 1971). However, the fact that the intensely Cr-PTA reactive band between enamel and ameloblasts did not show any particular reaction with PA-silver seems to imply that the carbohydrate composition of the Cr-PTA reactive band might be different from that of the surface layer of enamel which did get stained with PA-silver.…”

A cytochemical study of gl.ycocalyx and the membrane cholesterol of rat glomerular podocytes.

“…and Kanto Chem.). Ultrathin sections were cut by a Porter-Blum MT-1 ultramicrotome, then mounted on collodion coated nickel grids and stained with 1% phosphotungstic acid in 10% chromic acid (Cr-PTA) for 5 min (RAMBOURG et al, 1969). In order to identify Cr-PTA reactive sites, some sections were stained with uranyl acetate and lead citrate.…”

A Polysaccharide-Containing Cell Coat on Keratinizing Cells of the Romney Wool Follicle

“…However, the possible presence of specifically stained cytoplasmic granules may be significant. It is possible that a mechanism involving the Golgi apparatus, similar to that thought to operate in the epidermis (Matoltsy and Parakkal 1965) and intestinal columnar cells (Rambourg, Hernandez, and Leblond 1969), is present in the wool follicle.…”