Investigations of the Functional Expression of SLC6A14 in Non-CF and CF Airway Epithelial Cells
Year: 2014
Type: dissertation
Abstract: Recent studies have found single nucleotide polymorphisms significantly associated with more severe cystic fibrosis (CF) in the promoter region of the SLC6A14 gene, encoding a NaCl-dependent neutral and cationic amino acid transporter highly expressed in the lung. In a CF human bronchial epithelial cell line (CFBE41o-) transfected with wild-type- (CFBEwt) or dF505-CFTR (CFBEdf), 3H-arginine uptake followed Michaelis-Menten kinetics (CFBEwt Kt=95.01±14.05, CFBEdf Kt=72.8±16.14). Competitive inhibition experiments demonstrated function attributable to SLC6A14 and cationic amino acid transporter system y+ in both cells. Intracellular and transepithelial transport of 3H-arginine in non-CF and CF primary human airway cells was significantly reduced (~50%) in 0 Na+ or presence of a SLC6A14 blocker, alpha-methyl-DL-tryptophan (alpha-MT) apically, indicative of SLC6A14. There was no difference in SLC6A14 function between CF and non-CF cells, or CFBEwt and CFBEdf cells. These protocols allow isolation of SLC6A14 function in airway cells by competitive inhibition, Na+ dependence, and alpha -MT sensitivity.
Source: TSpace (University of Toronto)
Author: Clifford Andrew Lloyd-Kuzik
Language: English
Subfield: Pulmonary and Respiratory Medicine
Field: Medicine
Domain: Health Sciences
Open Access status: green
OpenAlex ID: W3136405326