Appearance of surfactant-like particles in apical medium of Caco-2 cells may occur via tight junctions.
Intestinal alkaline phosphatase (AP) is secreted by Caco-2 cells bound to surfactant-like particles (SLP), which can be localized by electron microscopy to the basolateral space and the intestinal lumen, especially over tight junctions. To investigate the hypothesis that SLP are secreted basolaterally and enter the lumen through the tight junction, Caco-2 cells were incubated with agents known to increase permeability at tight junctions. Cytochalasin D and phorbol 12-myristate 13-acetate increased Caco-2 cell monolayer permeability and the appearance of particles in apical medium two- to threefold, as monitored by mannitol movement and AP activity, respectively. Blocking the apical secretory pathway by nocodazole or colchicine had no effect on either parameter. Estimation of SLP content demonstrated an increase in apical media particles similar to that determined by AP activity. Quantitative image analysis established that apical SLP content increased 4-10 times, whereas total cell particle content remained unchanged. These data indicate that SLP may be secreted initially into the basolateral space and then transported to the intestinal lumen through the tight junctions.
Engle
M J
MJ
Gastroenterology Division, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Grove
M L
ML
Becich
M J
MJ
Mahmood
A
A
Alpers
D H
DH
D000230:Adenocarcinoma; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D000906:Antibodies; D002460:Cell Line; D002462:Cell Membrane / Q000648:ultrastructure; D002463:Cell Membrane Permeability / Q000187:drug effects; D003110:Colonic Neoplasms; D015638:Cytochalasin D / Q000494:pharmacology; D006801:Humans; D006982:Hypertonic Solutions / Q000494:pharmacology; D007365:Intercellular Junctions / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure*; D007421:Intestine, Small / Q000648:ultrastructure; D008854:Microscopy, Electron; D015388:Organelles / Q000187:drug effects / Q000648:ultrastructure*; D011817:Rabbits / Q000276:immunology; D011887:Raffinose; D013395:Sucrose; D013501:Surface-Active Agents; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013997:Time Factors; D014407:Tumor Cells, Cultured
D000906:Antibodies; D006982:Hypertonic Solutions; D013501:Surface-Active Agents; D015638:Cytochalasin D; D013395:Sucrose; D000469:Alkaline Phosphatase; D011887:Raffinose; D013755:Tetradecanoylphorbol Acetate