Intestinal apoproteins during fat absorption

To compare the roles of apolipoprotein (Apo) A-I, B, and E (or arginine-rich apoprotein, ARP) in the intracellular production of intestinal chylomicrons (and/or VLDL), these apoproteins were localized in rat intestinal mucosa by the light microscope method of indirect immunofluorescence. In addition, tissue levels of ApoA-I and ApoB were measured during fat absorption by radioimmunoassay. Antisera were produced using ApoA-I isolated from rat plasma high density lipoprotein, and ApoB and ARP from plasma VLDL by column chromatography. The apoproteins yielded single bands on polyacrylamide disc gel electrophoresis in urea and in sodium dodecyl sulfate. Anti-apoprotein antisera were produced in rabbits. These antisera appeared to be monospecific on double-antibody immunoprecipitation of 125I-labeled apoproteins. In fasted animals granular staining of ApoA-I was noted in the supranuclear (Golgi) regions of epithelial cells in the top third of the villus. At 30 min, when fat droplets were seen in the supranuclear cytoplasm of the cells along the top two-thirds of the villus, intense ApoA-I staining surrounded droplets in the cytoplasm. At later times when epithelial cells and lamina propria both contained fat droplets, bright ApoA-I stain surrounded many droplets in the supranuclear cytoplasm of cells and in the lamina propria. Over the same period of time, tissue levels of ApoA-I rose 10-fold. The distribution and time-course of ApoB staining was nearly identical with that of ApoA-I. Concomitantly, tissue ApoB levels doubled. By contrast, in fasting rat intestine, staining of ARP was sparse, punctate, and confined to the lower quarter of the villus. After fat feeding, stained droplets were seen only in the lamina propria near the base of the villus even though abundant ARP was found in cells along most of this length of the villus. Stain was never seen to surround any droplets inside cells. Thus, ApoA-I and ApoB appeared to participate in the intracellular assemply of lipoproteins in gut, whereas ARP did not, although ARP was found within mucosal cells. Liver and intestine differed in their stainable contents of ApoA-I and ARP. Whereas intestine stained heavily for ApoA-I and lightly for ARP, liver stained heavily for ARP and lightly for ApoA-I. Both organs stained for ApoB. These findings suggest that there may be some quantitative "specialization" of the two organs which secrete lipoproteins.