Insulin binding and fluid-phase endocytosis stimulation in the mouse neuroblastoma cell line 41A3.
As well as many other hormones and growth factors, insulin is known to influence several processes in the CNS; its specific effects, however, are still poorly understood. Neuroblastoma cell lines represent a useful experimental system for the analysis of the insulin-specific effect on neurons, in the absence of possible regulatory mechanisms elicited by other neuronal/glial cells and/or soluble factors. The expression and the binding properties of insulin receptors, as well as the insulin effects on both membrane fluidity and cell surface architecture, have been investigated in 41A3 mouse neuroblastoma cells, by radioligand-binding fluorescence spectroscopy and scanning electron microscopy, respectively the same cells, insulin-induced modifications on cytoskeletal organisation also have been studied. Binding studies were performed using 125I-insulin, while the cationic fluorescent probe trimethylammonium 1,6-diphenyl-1,3,5-hexatriene was used for biophysical investigations. The results presented in this paper provide evidence that insulin interacts with 41A3 neuroblastoma cells through a receptor-mediated mechanism and that, in these cells, insulin binding modifies the cell surface morphology and stimulates endocytosis.
Sartori
C
C
Department of Cellular and Developmental Biology, University of Rome La Sapienza, Italy.
Silvestroni
L
L
Stefanini
S
S
Tocco
G A
GA
D000818:Animals; D001667:Binding, Competitive / Q000502:physiology; D003911:Dextrans / Q000493:pharmacokinetics; D004161:Diphenylhexatriene / Q000031:analogs & derivatives; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology*; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000493:pharmacokinetics; D005456:Fluorescent Dyes; D006735:Horseradish Peroxidase / Q000493:pharmacokinetics; D007328:Insulin / Q000378:metabolism* / Q000494:pharmacology; D051379:Mice; D008855:Microscopy, Electron, Scanning; D009447:Neuroblastoma; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure
D003911:Dextrans; D005456:Fluorescent Dyes; D007328:Insulin; C015219:fluorescein isothiocyanate dextran; D004161:Diphenylhexatriene; C033628:1-(4-(trimethylamino)phenyl)-6-phenylhexa-1,3,5-triene; D006735:Horseradish Peroxidase; D016650:Fluorescein-5-isothiocyanate