Glutamine supports recovery from loss of transepithelial resistance and increase of permeability induced by media change in Caco-2 cells.
Recent evidence suggests that the conditionally essential amino acid glutamine is important for intestinal barrier function. However, the mechanism remains undefined. To determine the effects of glutamine on permeability of intestinal epithelial cell monolayers, Caco-2 cells were grown on membrane filters and exposed to 4 mmol/L sodium butyrate in order to rapidly achieve high levels of alkaline phosphatase and high transepithelial resistance as seen in functionally mature enterocytes. A standard method of medium exchange consisting of removal and replacement resulted in a catastrophic loss of transepithelial resistance and increase of mannitol and dextran fluxes that required 2-4 hrs and protein synthesis to recover. The effect was attributed to exposure of the upper monolayer surface to atmosphere and could be avoided by refeeding by incremental perfusion. Spontaneously-differentiated Caco-2 monolayers were resistant to this stress. This novel stress test was employed as a sensitive assay for the requirement of glutamine for monolayer transepithelial resistance and mannitol permeability. Pre-stress glutamine availability was more important than Gln-availability during the recovery phase. Thus the transepithelial resistance and permeability of butyrate-induced monolayers is dynamically-regulated in response to atmospheric exposure, by a mechanism that depends on threshold levels of glutamine availability.
Li
Nan
N
Department of Pediatrics, University of Florida, College of Medicine, Gainesville, Florida 32610, USA.
DeMarco
Vincent G
VG
West
Christopher M
CM
Neu
Josef
J
D000469:Alkaline Phosphatase / Q000378:metabolism; D002087:Butyrates / Q000494:pharmacology; D018938:Caco-2 Cells; D002463:Cell Membrane Permeability / Q000187:drug effects*; D003470:Culture Media; D003911:Dextrans / Q000378:metabolism; D017097:Electric Impedance; D004847:Epithelial Cells / Q000502:physiology / Q000648:ultrastructure; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives* / Q000378:metabolism; D005973:Glutamine / Q000494:pharmacology*; D006801:Humans; D007422:Intestines / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D008353:Mannitol / Q000378:metabolism
D002087:Butyrates; D003470:Culture Media; D003911:Dextrans; C015219:fluorescein isothiocyanate dextran; D005973:Glutamine; D008353:Mannitol; D000469:Alkaline Phosphatase; D016650:Fluorescein-5-isothiocyanate