Quantitative and qualitative characterization of apolipoprotein B containing lipoproteins produced by the visceral rat yolk sac in two different in vitro systems: organ culture and isolated epithelial cells in suspension culture.
Tissue pieces as well as isolated epithelial cells taken from visceral rat yolk sacs at the 18th day of gestation were able to synthesize and to secrete apo B containing lipoproteins floating in the density ranges of VLDL, IDL and LDL. In all three density classes only the high molecular weight apo B was detectable. VLDL secreted from yolk sac tissue or isolated epithelial cells lacked apo E and apo C. Studies with cycloheximide revealed that about 77% of the particles was delivered from a pool of preformed lipoproteins. Evidence was given that also de novo-synthesis of apo B containing lipoproteins took place in the tissue segments and the isolated epithelial cells. Most of apo B mass (90%) and of apo B radioactivity (60%) secreted by the tissue pieces of the visceral yolk sac floated in the density range 1.020-1.064 g/ml, the remainder being found in the d < or = 1.020 g/ml fraction. In contrast, only 40% of apo B mass and 45% of apo B radioactivity delivered from isolated epithelial cells belonged to the d = 1.020-1.064 g/ml fraction. LDL released from yolk sacs and isolated epithelial cells contained more triacylglycerols (41% vs. 25%) and had a larger mean diameter (24 nm vs. 21.8 nm) than those obtained from fetal rat serum, whereas the comparatively small VLDL produced in vitro (mean diameter = 34 nm) contained less triacylglycerols (46% vs. 60.5%) and more protein (20% vs. 10.2%) in comparison with fetal serum VLDL (mean diameter = 42.3 nm). Incubation experiments with [125I]VLDL led to the conclusion that the lipase secreted by yolk sac tissue into the medium could not be responsible for the conversion of VLDL into LDL, thus supporting our view of a direct LDL secretion by visceral rat yolk sacs.
Plonné
D
D
Institute of Pathobiochemistry, Friedrich-Schiller University Jena, Germany.
Heller
H
H
Kahlert
U
U
Dargel
R
R
D000818:Animals; D001055:Apolipoproteins B / Q000032:analysis*; D002478:Cells, Cultured; D004848:Epithelium / Q000378:metabolism; D005260:Female; D005865:Gestational Age; D008049:Lipase / Q000378:metabolism; D008074:Lipoproteins / Q000032:analysis* / Q000378:metabolism; D053419:Lipoproteins, IDL; D008077:Lipoproteins, LDL / Q000032:analysis / Q000378:metabolism; D008079:Lipoproteins, VLDL / Q000032:analysis / Q000378:metabolism; D008297:Male; D009924:Organ Culture Techniques; D010316:Particle Size; D011247:Pregnancy; D051381:Rats; D017208:Rats, Wistar; D015017:Yolk Sac / Q000378:metabolism*
D001055:Apolipoproteins B; D008074:Lipoproteins; D053419:Lipoproteins, IDL; D008077:Lipoproteins, LDL; D008079:Lipoproteins, VLDL; D008049:Lipase