Histochemical evidence for the differential surface labeling, uptake, and intracellular transport of a colloidal gold-labeled insulin complex by normal human blood cells.
Open Access
- 1 October 1981
- journal article
- research article
- Published by SAGE Publications in Journal of Histochemistry & Cytochemistry
- Vol. 29 (10), 1137-1149
- https://doi.org/10.1177/29.10.6271866
Abstract
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.This publication has 23 references indexed in Scilit:
- Characteristics of human erythrocyte insulin receptorsDiabetes, 1978
- Direct visualization of binding, aggregation, and internalization of insulin and epidermal growth factor on living fibroblastic cellsProceedings of the National Academy of Sciences, 1978
- Intracellular Translocation of Iodine-125-Labeled Insulin: Direct Demonstration in Isolated HepatocytesScience, 1978
- 125I-insulin binding to cultured human lymphocytes. Initial localization and fate of hormone determined by quantitative electron microscopic autoradiography.JCI Insight, 1978
- The monocyte as a model for the study of insulin receptors in manDiabetologia, 1977
- Adsorption of horseradish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of concanavalin A, wheat germ agglutinin and goat anti-human immunoglobulin G on cell surfaces at the electron microscopic level: a new method, theory and application.Journal of Histochemistry & Cytochemistry, 1977
- Cellular uptake and nuclear binding of insulin in human cultured lymphocytes: evidence for potential intracellular sites of insulin action.Proceedings of the National Academy of Sciences, 1977
- Colloidal gold, a useful marker for transmission and scanning electron microscopy.Journal of Histochemistry & Cytochemistry, 1977
- Binding and degradation of insulin by human peripheral granulocytes. Demonstration of specific receptors with high affinity.Journal of Biological Chemistry, 1976
- INSULIN BINDING OF ACUTE LYMPHOCYTIC LEUKEMIA-CELLS1976