An electron microscope autoradiographic study of the carbohydrate recognition systems in rat liver. I. Distribution of 125I-ligands among the liver cell types

Ann Louise Hubbard, G. Wilson, G. Ashwell, H. Stukenbrok

Research output: Contribution to journalArticle

Abstract

Electron microscope autoradiography was used to study the cellular localization of seven glycoproteins rapidly cleared from the circulating plasma of rats and taken up by the liver. 1 and 15 min after intravenous administration of the 125I-glycoproteins, livers were fixed in situ by perfusion and processes for autoradiography. Autoradiographic grains in the developed section were found to represent the intact 125I-ligand. A quantitative analysis of the distribution and concentration (density) of autoradiographic grains over the three major cell types of the liver was then performed. Three molecules, asialo-fetuin, asialo-orosomucoid, and lactosaminated RNase A dimer, the oligosaccharide chains of which terminate in galactose residues, were bound and internalized almost exclusively (>90%) by hepatocytes. Conversely, four molecules, the oligosaccharide chains of which terminate in either N-acetyl-glucosamine (agalacto-orosomucoid) or mannose (ahexosamino-orosomucoid, preputial β-glucuronidase, and mannobiosaminated RNase A dimer), were specifically bound and internalized by cells lining the blood sinusoids-that is, by Kupffer cells and endothelial cells. Endothelial cells were two to six times more active (on a cell volume basis) than were Kupffer cells in the internalization of these four 125I-ligands. Mannose and N-acetylglucosamine-terminated glycoproteins competed with each other for uptake into either endothelial cells or Kupffer cells, indicating that a single system recognized mannose or N-acetyl-glucosamine residues. Finally, agalacto-orosomucoid and ahexosamino-orosomucoid were also associated with hepatocytes, but competition experiments utilizing excess asialo-orosomucoid demonstrated that residual galactosyl residues were responsible for this association.

Original languageEnglish (US)
Pages (from-to)47-64
Number of pages18
JournalJournal of Cell Biology
Volume83
Issue number1
StatePublished - 1979
Externally publishedYes

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Kupffer Cells
Mannose
Pancreatic Ribonuclease
Glycoproteins
Endothelial Cells
Glucosamine
Carbohydrates
Electrons
Ligands
Autoradiography
Oligosaccharides
Hepatocytes
Liver
Acetylglucosamine
Glucuronidase
Galactose
Cell Size
Intravenous Administration
Blood Cells
Perfusion

ASJC Scopus subject areas

  • Cell Biology

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An electron microscope autoradiographic study of the carbohydrate recognition systems in rat liver. I. Distribution of 125I-ligands among the liver cell types. / Hubbard, Ann Louise; Wilson, G.; Ashwell, G.; Stukenbrok, H.

In: Journal of Cell Biology, Vol. 83, No. 1, 1979, p. 47-64.

Research output: Contribution to journalArticle

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