Glucocorticoid regulation and glycosylation of mouse intestinal type IIb Na-Pi cotransporter during ontogeny

Abstract

We sought to characterize expression of an apically expressed intestinal Na-Pi cotransporter (Na-Pi-IIb) during mouse ontogeny and to assess the effects of methylprednisolone (MP) treatment. In control mice, Na-Pi uptake by intestinal brush-border membrane vesicles was highest at 14 days of age, lower at 21 days, and further reduced at 8 wk and 8–9 mo of age. Na-Pi-IIb mRNA and immunoreactive protein levels in 14-day-old animals were markedly higher than in older groups. MP treatment significantly decreased Na-Pi uptake and Na-Pi-IIb mRNA and protein expression in 14-day-old mice. Additionally, the size of the protein was smaller in 14-day-old mice. Deglycosylation of protein from 14-day-old and 8-wk-old animals with peptide N-glycosidase reduced the molecular weight to the predicted size. We conclude that intestinal Na-Pi uptake and Na-Pi-IIb expression are highest at 14 days and decrease with age. Furthermore, MP treatment reduced intestinal Na-Pi uptake approximately threefold in 14-day-old mice and this reduction correlates with reduced Na-Pi-IIb mRNA and protein expression. We also demonstrate that Na-Pi-IIb is an N-linked glycoprotein and that glycosylation is age dependent.