This chapter describes the use of isolated, perfused liver in the studies of receptor-mediated endocytosis. The chapter focuses on the asialoglycoprotein (ASGP) recognition system in rat liver, which mediates the clearance of glycoproteins having galactose- or N-acetylgalac- tosamine-terminating carbohydrate side chains. The dynamics of the ligand pathway in vivo, in isolated hepatocytes, and in the isolated perfused liver is studied. The isolated perfused liver has been used in a variety of forms by many investigators for the study of a number of liver functions. The liver perfusion system comprises three primary components: (1) a temperature-regulation system; (2) a pump; and (3) gas-permeable fibers mounted in a beaker assembly. The chapter also illustrates a step-by-step procedure for isolating a rat liver, procedures for performing temperature-shift experiments, and presents an outline of the method used to label selectively the contents of endocytic compartments, using the asialoglycoprotein system as a model. The selected functions of the perfused liver that relate to endocytosis are examined.
ASJC Scopus subject areas
- Molecular Biology