Inhibition of apical but not basolateral endocytosis of ricin and folate in Caco-2 cells by cytochalasin D.
Apical and basolateral endocytic pathways in polarised Caco-2 cells were investigated by following the uptake, recycling and transcytosis of the galactose-binding protein toxin ricin, as a membrane marker. Differences in the extent and kinetics of lectin uptake, recycling and transcytosis were observed at the apical and basolateral domains and altered with the age of the cell monolayer. Treatment of polarised Caco-2 cells with cytochalasin D showed a domain-specific, concentration-dependent inhibition of apical endocytosis of ricin. Inhibition of apical endocytosis by cytochalasin D was not due to a gross change in brush border morphology, although actin stress fibres within the cell body were disrupted. It is not clear whether inhibition of apical endocytosis in polarized epithelial cells by cytochalasin D is caused simply by disruption of a mechanochemical motor involving microvillar actin filaments. The cytochalasin D effect was also observed when measuring uptake of folate, suggesting apical domain-specific inhibition of caveolar, as well as clathrin-mediated, endocytic routes.
Department of Clinical Biochemistry, University of Cambridge, Addenbrooke's Hospital, UK.
M R
MR
Jackman
W
W
Shurety
J A
JA
Ellis
J P
JP
Luzio
D008841:Actin Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D000230:Adenocarcinoma / Q000473:pathology*; D000818:Animals; D002460:Cell Line; D016764:Cell Polarity; D003110:Colonic Neoplasms / Q000473:pathology*; D015638:Cytochalasin D / Q000494:pharmacology*; D003864:Depression, Chemical; D004285:Dogs; D004705:Endocytosis / Q000187:drug effects*; D005492:Folic Acid / Q000378:metabolism*; D006801:Humans; D007668:Kidney; D008871:Microvilli / Q000378:metabolism / Q000648:ultrastructure; D009363:Neoplasm Proteins / Q000378:metabolism; D012276:Ricin / Q000378:metabolism*; D014407:Tumor Cells, Cultured / Q000187:drug effects
D009363:Neoplasm Proteins; D015638:Cytochalasin D; D012276:Ricin; D005492:Folic Acid