Morphologic demonstration of receptor-mediated endocytosis of epidermal growth factor by isolated bile duct epithelial cells

Motoyasu Ishii, Benjamin Vroman, Nicholas F La Russo

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Abstract

It was recently shown that intrahepatic bile duct epithelial cells in situ or after isolation from rat liver have coated pits and vesicles, suggesting that they participate in receptor-mediated endocytosis. Therefore, using a morphologic approach and epidermal growth factor coupled to horseradish peroxidase or colloidal gold as probes, we studied freshly isolated or short-term cultured intrahepatic bile duct epithelial cells prepared from normal rat liver to determine if they participate in receptor-mediated endocytosis. Immunoelectron microscopy using a monoclonal antibody against the epidermal growth factor receptor was also used to examine for the presence of the growth factor receptor on the cells. Immediately after isolation, the cells did not internalize either epidermal growth factor-horseradish peroxidase or epidermal growth factor-colloidal gold; no growth factor receptor could be shown on these cells by immunocytochemistry, either. In contrast, cells cultured for 24 h bound and internalized both epidermal growth factor-horseradish peroxidase and epidermal growth factor-colloidal gold at 37 °C and showed growth factor receptors diffusely distributed on the plasma membrane. When cultured cells exposed to epidermal growth factor-colloidal gold were fixed with glutaraldehyde containing saponin and tannic acid, colloidal gold particles were observed in coated pits and in coated and uncoated vesicles. Preincubation of cultured cells with native epidermal growth factor completely blocked the internalization of both epidermal growth factor-horseradish peroxidase and epidermal growth factor-colloidal gold. When rat liver was stained in situ for epidermal growth factor receptor, reaction product was observed by immunoelectron microscopy exclusively on the basal surface of the plasma membrane of the intrahepatic bile duct epithelial cells. These results indicate that bile duct epithelial cells internalize epidermal growth factor by endocytosis via coated pits containing receptors localized in situ exclusively to the basal domain of their plasma membranes. The data demonstrate for the first time that intrahepatic bile duct epithelial cells participate in receptor-mediated endocytosis and raise the possibility that they are a target for epidermal growth factor.

Original languageEnglish (US)
Pages (from-to)1284-1291
Number of pages8
JournalGastroenterology
Volume98
Issue number5 PART 1
DOIs
StatePublished - 1990

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Endocytosis
Bile Ducts
Epidermal Growth Factor
Epithelial Cells
Gold Colloid
Intrahepatic Bile Ducts
Growth Factor Receptors
Horseradish Peroxidase
Coated Vesicles
Cultured Cells
Immunoelectron Microscopy
Cell Membrane
Epidermal Growth Factor Receptor
Liver
Cell Separation
Tannins
Saponins
Glutaral
Immunohistochemistry
Monoclonal Antibodies

ASJC Scopus subject areas

  • Gastroenterology

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Morphologic demonstration of receptor-mediated endocytosis of epidermal growth factor by isolated bile duct epithelial cells. / Ishii, Motoyasu; Vroman, Benjamin; La Russo, Nicholas F.

In: Gastroenterology, Vol. 98, No. 5 PART 1, 1990, p. 1284-1291.

Research output: Contribution to journalArticle

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