Differential regulation of cystic fibrosis transmembrane conductance regulator and Na+,K+-ATPase in gills of striped bass, Morone saxatilis: effect of salinity and hormones

Abstract

Effects of salinity and hormones on cystic fibrosis transmembrane conductance regulator (CFTR) and alpha-subunit Na(+),K(+) -ATPase (alpha-NKA) mRNA (analysed by semi-quantitative PCR) and protein expression (analysed by western blotting and immunocytochemistry) were investigated in gills of striped bass. Freshwater (FW) to seawater (SW) transfer induced a disturbance in serum [Na(+)]. Gill CFTR protein, mRNA level and Na(+),K(+) -ATPase activity were unaffected by SW transfer, whereas alpha-NKA mRNA increased after transfer. CFTR immunoreactivity was observed in large cells in FW and SW gill filaments at equal intensity. Cortisol decreased serum [Na(+)] in FW fish, but had no effect on gill Na(+),K(+) -ATPase activity, alpha-NKA and CFTR mRNA levels. Incubation of gill tissue with cortisol (24 h, >0.01 micro g/ml) and epidermal growth factor (EGF 10 micro g/ml) decreased CFTR mRNA levels relative to pre-incubation and control levels. CFTR expression was unaffected by IGF-I (10 micro g/ml). alpha-NKA mRNA levels decreased by 50% after 24 h control incubation; it was slightly stimulated by cortisol and unaffected by IGF-I and EGF. In isolated gill cells, phosphorylation of extracellular-regulated kinase (ERK) 1/2 was stimulated by EGF but not affected by IGF-I. This study is the first to report a branchial EGF response and to demonstrate a functional ERK 1/2 pathway in the teleost gill. In conclusion, CFTR and Na(+),K(+) -ATPase are differentially regulated by salinity and hormones in gills of striped bass, despite the putative involvement of both in salt excretion.