The use of lectins and cholera toxin for the detection of surface carbohydrates of cultured neurons and neuroblastoma.
Conjugates of horseradish peroxidase with the lectins ricin (d-galactose), wheat germ agglutinin (N-acetylglucosamine), phytohemagglutinin (N-acetylgalactosamine), and with cholera toxin (GM1 ganglioside) were used for a cytochemical detection of corresponding termin al carbohydrates, or glycolipids on cell surfaces of cultured neurons and neuroblastoma cells. Cells were labeled at 4 degrees C with the above ligands and their adsorptive endocytosis was studied after incubations at 37 degrees C in a medium free of ligand. Peroxidase was detected by the method of Graham and Karnovsky (J. Histochem. Cytochem. 14:291, 1966). Lectins and cholera toxin underwent endocytosis in cisternae and vesicles of GERL (Golgi-Endoplasmic Reticulum-Lysosome). We suggest that GERL is the primary ercipieint of adsorptively endocytosed plasma membrane "receptor"-ligand complexes which are thus degraded or possibly reutilized (recycling). Wheat germ agglutinin-horseradish peroxidase conjugates used in vivo for studies of retrograde axonal transport were significantly more sensitive than free horseradish peroxidase.
D002241:Carbohydrates / Q000302:isolation & purification*; D002478:Cells, Cultured; D002772:Cholera Toxin; D004705:Endocytosis; D037102:Lectins; D009447:Neuroblastoma / Q000032:analysis*; D009474:Neurons / Q000032:analysis*; D013499:Surface Properties
D002241:Carbohydrates; D037102:Lectins; D002772:Cholera Toxin