Histochemical evidence for the differential surface labeling, uptake, and intracellular transport of a colloidal gold-labeled insulin complex by normal human blood cells.
A colloidal gold-labeled insulin-bovine serum albumin (GIA) reagent has been developed for the ultrastructural visualization of insulin binding sites on the cell surface and for tracing the pathway of intracellular insulin translocation. When applied to normal human blood cells, it was demonstrated by both visual inspection and quantitative analysis that the extent of surface labeling, as well as the rate and degree of internalization of the insulin complex, was directly related to cell type. Further, the pathway of insulin (GIA) transport via round vesicles and by tubulo-vesicles and saccules and its subsequent fate in the hemic cells was also related to cell variety. Monocytes followed by neutrophils bound the greatest amount of labeled insulin. The majority of lymphocytes bound and internalized little GIA, however, between 5-10% of the lymphocytes were found to bind considerable quantities of GIA. Erythrocytes rarely bound the labeled insulin complex, while platelets were noted to sequester large quantities of the GIA within their extracellular canalicular system. GIA uptake by the various types of leukocytic cells appeared to occur primarily by micropinocytosis and by the direct opening of cytoplasmic tubulo-vesicles and saccules onto the cell surface in regions directly underlying surface-bound GIA. Control procedures, viz., competitive inhibition of GIA labeling using an excess of unlabeled insulin in the incubation medium, preincubation of the GIA reagent with an antibody directed toward porcine insulin, and the incorporation of 125I-insulin into the GIA reagent, indicated the specificity and selectivity of the GIA histochemical procedure for the localization of insulin binding sites.
D000328:Adult; D001692:Biological Transport; D001792:Blood Platelets / Q000378:metabolism / Q000648:ultrastructure; D004912:Erythrocytes / Q000378:metabolism; D005215:Fasting; D006048:Gold Colloid, Radioactive / Q000378:metabolism*; D006651:Histocytochemistry; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D007962:Leukocytes / Q000378:metabolism* / Q000648:ultrastructure; D010873:Pinocytosis; D011972:Receptor, Insulin / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000378:metabolism*
D006048:Gold Colloid, Radioactive; D007328:Insulin; D012710:Serum Albumin, Bovine; D011972:Receptor, Insulin