Transcytosis of pancreatic bile salt-dependent lipase through human Int407 intestinal cells.
In previous studies, we have shown that the bile-salt-dependent-lipase (BSDL), secreted by pancreatic acinar cells and secreted into the duodenal lumen, can be transcytosed through intestinal cells up to the lamina propria. In this study, we used an in vitro system to provide insights into the apical to basolateral transport of BSDL, across the intestinal barrier. The Int407 human epithelial cell line, grown under conditions that optimize polarity, was used as a tight epithelium model. We attempted to delineate uptake mechanisms and the transcytotic pathway followed by this pancreatic enzyme within the intestinal Int407 cells, which do not produce BSDL. When added to the apical reservoir of Transwell-grown Int407 cells, BSDL was shown to first interact with the apical membrane. Further, BSDL forms clusters that are internalized via clathrin-coated pits. Following endocytosis, BSDL is directed to a nocodazole- and colchicin-sensitive multivesicular compartment. Interestingly, this protein transits through the Golgi apparatus, where it was found to colocalize with the KDEL retrieval-receptor. Finally, enzymatically active intact BSDL was released at the basolateral membrane level. This is the first demonstration for an apical-to-basolateral transcytotic pathway of a secreted pancreatic digestive enzyme through polarized intestinal cells.
Bruneau
N
N
Unité de Recherche de Physiopathologie des cellules épithéliales, INSERM U-559, Marseilles, France.
Nganga
A
A
Bendayan
M
M
Lombardo
D
D
D000818:Animals; D002460:Cell Line; D016764:Cell Polarity; D002966:Clathrin / Q000378:metabolism; D003034:Coated Pits, Cell-Membrane / Q000378:metabolism; D004386:Duodenum; D004705:Endocytosis / Q000502:physiology; D005089:Exocytosis / Q000502:physiology; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007413:Intestinal Mucosa / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008854:Microscopy, Electron; D010179:Pancreas / Q000166:cytology / Q000201:enzymology; D021381:Protein Transport / Q000502:physiology*; D051381:Rats; D018000:Receptors, Peptide / Q000378:metabolism; D002787:Sterol Esterase / Q000378:metabolism* / Q000494:pharmacology
D002966:Clathrin; C064459:KDEL receptor; D018000:Receptors, Peptide; C054784:bile salt-stimulated lipase; D002787:Sterol Esterase