Abstract
BackgroundSmall interfering RNA (siRNA) against αENaC (α-subunit of the epithelial Na channel) and CFTR (cystic fibrosis transmembrane conductance regulator) was used to explore ENaC and CTFR function in newborn rat lungs.MethodsTwenty-four hours after trans-thoracic intrapulmonary (ttip) injection of siRNA-generating plasmid DNA (pSi-0, pSi-4, or pSi-C2), we measured CFTR and ENaC expression, extravascular lung water, and mortality.ResultsαENaC and CFTR mRNA and protein decreased by ~80% and ~85%, respectively, following αENaC and CFTR silencing. Extravascular lung water and mortality increased after αENaC and CFTR-silencing. In pSi-C2-transfected isolated DLE cells there were attenuated CFTR mRNA and protein. In pSi-4-transfected DLE cells αENaC mRNA and protein were both reduced. Interestingly, CFTR-silencing also reduced αENaC mRNA and protein. αENaC silencing, on the other hand, only slightly reduced CFTR mRNA and protein.ConclusionThus, ENaC and CFTR are both involved in the fluid secretion to absorption conversion around at birth.
Highlights
Small interfering RNA against αENaC (α-subunit of the epithelial Na channel) and CFTR was used to explore epithelial Na channels (ENaC) and CTFR function in newborn rat lungs
ENaC and CFTR are both involved in the fluid secretion to absorption conversion around at birth
Lung CFTR mRNA during development We determined if CFTR transcription changed during early postnatal development
Summary
Small interfering RNA (siRNA) against αENaC (α-subunit of the epithelial Na channel) and CFTR (cystic fibrosis transmembrane conductance regulator) was used to explore ENaC and CTFR function in newborn rat lungs. Fetal lungs are filled with fluid that is produced and secreted by the pulmonary epithelium and linked to Nacoupled Cl secretion in utero. This fluid must be rapidly removed at birth for adequate gas exchange across the alveolar epithelium-endothelium at birth to occur. Studies of cultured alveolar type II epithelial cells suggest that cystic fibrosis transmembrane conductance regulator-(CFTR)-mediated Cl transport may (page number not for citation purposes)
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