The study of biogenetic pathways using a perfusion technique containing perfluorochemicals.
A liver perfusion system was assembled and adapted for pulse labelling studies. The perfusion medium was prepared by emulsifying perfluorotributylamine and Pluronic F 68 in a CO2 atmosphere using a sonicator. Ribosome-rich and ribosome-poor rough microsomes, smooth microsomes and Golgi membranes could be prepared from perfused livers with a good purity and recovery as from non-perfused livers. The subfractionation technique used was simple and involved slight modifications of the methods described earlier by Eriksson (1973) and Ehrenreich et al. (1973). The specific activity of NADPH-cytochrome c reductase in microsomes and of UDP-galactosyltransferase in Golgi membranes from perfused and non-perfused livers were identical. The specific activity of glucose-6-phosphatase in microsomes was slightly decreased after perfusion, but the membrane permeability barrier to glucose-6-phosphate remained intact. The granulated microsomal fractions from perfused liver had a somewhat reduced number of membrane-bound ribosomes. The system developed proved useful in studies of the synthesis and intracellular transport of albumin. The technique should also be suitable for use in studies of membrane biogenesis.
D000818:Animals; D002463:Cell Membrane Permeability; D004579:Electron Transport; D005466:Fluorocarbons; D007425:Intracellular Membranes / Q000378:metabolism; D008099:Liver / Q000378:metabolism*; D008297:Male; D010477:Perfusion / Q000379:methods; D051381:Rats; D013347:Subcellular Fractions / Q000378:metabolism
D005466:Fluorocarbons