Neurotransmitter-stimulated ion transport across cultured bovine mammary epithelial cell monolayers

Abstract

Bovine mammary epithelial (BME-UV) and myoepithelial (BMM-UV) cell lines were acquired with the goal of developing an in vitro model of mammary epithelia for the study of ion transport. The bovine mammary cell lines were successfully cultured on commercially available permeable supports, and results suggest that mammary epithelial cells, but not myoepithelial cells, form tight junctions necessary to perform a barrier function. Electrogenic ion transport was not observed in basal conditions. Acute exposure to norepinephrine or forskolin caused prototypic increases in short circuit current accompanied by a reduction in transmural resistance indicative of anion secretion through a conductive pathway. Bumetanide and N-(4-methyphenylsulfonyl)-N′-(4-trifluoro-methylphenyl)urea, inhibitors of Na+/K+/Cl- cotransport and cystic fibrosis transmembrane conductance anion channels, respectively, reduced forskolin-stimulated ion transport. Amiloride, an inhibitor of epithelial sodium channels, had no effect on basal or forskolin-stimulated ion transport. However, naturally occurring and synthetic corticosteroids induced the expression of amiloride sensitive current indicative of sodium absorption. Chronic exposure to increased apical ionic strength and/or reduced carbohydrate concentration were associated with reduced transepithelial resistance although forskolin-stimulated ion transport was unaffected. These results demonstrate that neurotransmitters and steroid hormones act directly on bovine mammary epithelial cells to acutely and chronically modulate the volume and composition of their secretions. The in vitro system that we describe can be further exploited to characterize cellular and molecular mechanisms associated with mammary function in health and disease.