title: Glucocorticoid-induced formation of tight junctions in mouse mammary epithelial cells in vitro.
abstract: Phenotypically stable cultures of untransformed mouse mammary epithelial cells (denoted 31EG4) were established and utilized to investigate the lactogenic hormone (glucocorticoids, insulin, and prolactin) regulation of tight junction formation. When 31EG4 cells were grown on permeable supports for 4 days in medium containing the synthetic glucocorticoid dexamethasone and insulin, confluent cell monolayers obtained a transepithelial electrical resistance (TER) of 1000-3000 omega.cm2. In contrast, over the same time period, confluent monolayers treated with insulin or insulin and prolactin maintained a low TER (35-150 omega.cm2). Consistent with the formation of tight junctions, apical to basolateral paracellular permeability was decreased from 12% to 1% for [14C]mannitol and 3.3% to 0.3% for [3H]inulin when cells were cultured in dexamethasone. This effect of dexamethasone on TER required extracellular calcium, de novo protein synthesis, dose-dependently correlated with glucocorticoid receptor occupancy, and was not due to an increase in cell density. As shown by direct and indirect immunofluorescence microscopy, dexamethasone treatment did not modulate the production or location of filamentous actin, the tight junction protein ZO-1, or the cell adhesion protein E-cadherin. Our results suggest that glucocorticoids play a fundamental role in the function and maintenance of cell-cell contact in the mammary epithelia by inducing the formation of tight junctions.
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley.
authors.forename: K S
authors.initials: KS
authors.lastname: Zettl
authors.forename: M D
authors.initials: MD
authors.lastname: Sjaastad
authors.forename: P M
authors.initials: PM
authors.lastname: Riskin
authors.forename: G
authors.initials: G
authors.lastname: Parry
authors.forename: T E
authors.initials: TE
authors.lastname: Machen
authors.forename: G L
authors.initials: GL
authors.lastname: Firestone
mesh_terms: D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002999:Clone Cells; D003513:Cycloheximide / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology*; D004553:Electric Conductivity / Q000187:drug effects; D004848:Epithelium / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D007328:Insulin / Q000494:pharmacology; D007365:Intercellular Junctions / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D007700:Kinetics; D008321:Mammary Glands, Animal / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D008353:Mannitol / Q000378:metabolism; D051379:Mice; D011388:Prolactin / Q000494:pharmacology
chemical_list: D007328:Insulin; D008353:Mannitol; D003907:Dexamethasone; D011388:Prolactin; D003513:Cycloheximide; D002118:Calcium