Apolipoprotein B-100: immunolocalization and synthesis in human intestinal mucosa.
Despite the evidence that the human small intestine produces two separate species of apoB mRNA encoding for B-100 and B-48, there is a paucity of data concerning the expression of the latter form in this organ. Using a high resolution immunogold approach, with specific polyclonal antibodies and a panel of monoclonal antibodies (2D8, 3A10, 4G3), both forms of apoB (B-48 and B-100) were revealed over enterocytes of pediatric intestinal samples. Intense labeling was observed over microvilli, apical smooth membrane vesicles, multivesicular bodies, the basolateral membrane, as well as the trans Golgi region. Only low labeling was found over the rough endoplasmic reticulum (rER). Similar patterns of apoB distribution characterized both duodenal and jejunal regions. The presence of labeling over the Golgi apparatus and rER suggests a synthetic activity of both forms of apoB by the epithelial cells. To test this hypothesis, human intestine was incubated with [3H]leucine, homogenized, and subjected to immunoprecipitation for apoB. Immunoprecipitates contained radioactivity mainly in apoB-48 with relatively small amounts in apoB-100 when examined by NaDodSO4-polyacrylamide gel electrophoresis. These findings were further supported by the biochemical determination of apoB-100 and apoB-48 in chylomicron particles isolated from thoracic duct lymph of a human donor. Taken together, our data suggest that the human intestine is able to synthesize and to express the apoB-100.
Levy
E
E
Department of Nutrition, University of Montréal, Québec, Canada.
Rochette
C
C
Londono
I
I
Roy
C C
CC
Milne
R W
RW
Marcel
Y L
YL
Bendayan
M
M
D053299:Apolipoprotein B-100; D001055:Apolipoproteins B / Q000032:analysis* / Q000096:biosynthesis; D002648:Child; D002675:Child, Preschool; D002914:Chylomicrons / Q000378:metabolism; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D007223:Infant; D007413:Intestinal Mucosa / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D007421:Intestine, Small / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D008196:Lymph / Q000737:chemistry; D008297:Male; D008854:Microscopy, Electron
D053299:Apolipoprotein B-100; D001055:Apolipoproteins B; D002914:Chylomicrons