Fetal lung epithelial ion channels relocate in the cell membrane during late gestation

Near the end of gestation, the direction of ion and fluid flow across the alveolar epithelium rapidly changes from secretion to absorption. Thus, the relative cell membrane location of epithelial Na channels (ENaCs) and cystic fibrosis transmembrane regulator (CFTR) Cl channels during late fetal lung development and after maternal interleukin-1β (IL-1β) pretreatment was the focus of our study. Western blot analysis after sucrose gradient separation of caveolin-1-(Cav-1)-rich membrane regions (CRR) and Cav-1-poor membrane (non-CRR) revealed primary CRR ENaC localization at gestation day (GD) 61 in guinea pigs. Correlating with the natural induction of distal lung fluid absorption, ENaC appeared in the non-CRR cell membrane regions at GD68. Conversely, CFTR was present in the non-CRR cell membrane regions at GD61 and in the CRRs at GD68. IL-1β-induced conversion to distal lung fluid absorption at GD61 was associated with ENaC non-CRR presence and CFTR CRR presence, suggesting that relative ENaC and CFTR locations induced distal lung fluid absorption and decreased fluid secretion. Instilling fetal lungs with the CRR-disrupting agent methyl-β-cyclodextrin resulted in the conversion from lung fluid secretion to absorption and ENaC non-CRR presence at GD61. Coimmunoprecipitation of Cav-1 with α- and β-ENaC demonstrated reduced coimmunoprecipitation with increased GD and after IL-1β pretreatment. On the other hand, coimmunoprecipitation of Cav-1 with CFTR demonstrated increased coimmunoprecipitation with increasing GD and after IL-1β pretreatment. This concept may provide novel molecular mechanisms for the rapid transition from fetal distal lung fluid secretion to absorption in near-term lungs.