Modelling Mice with Alveolar Epithelial Cell-Specific Conditional Knockout of α1,6-Fucosyltransferase

Abstract

Aim: To create a mice model to better evaluate the relationship between α1,6-fucosyltransferase (Fut8) with lung diseases like pulmonary fibrosis and lung inflammation. We created a mice model with Fut8 gene conditional knockout in the alveolar epithelial cell. Method: Alveolar epithelial cell specific Fut8 conditional knockout (CKO) mouse model was established based on the Cre/Loxp (cyclization recombination enzyme/locus of X (cross)-overinP1) system. Mice genotypes were identified by polymerase chain reaction (PCR). Fut8 protein and messenger ribonucleic acid (mRNA) expression in lung tissues or alveolar epithelial cells were examined using RT-PCR (reverse transcription-polymerase chain reaction), western blot and immunofluorescence (IF). Lectin blots and IF were used to assess the core fucosylation in lung tissues and primary alveolar epithelial cells. Fut8 enzyme activity was tested by high performance liquid chromatography (HPLC). Changes of lung tissue morphology were evaluated by histological examination. Result: Fut8 protein and mRNA expression in lung tissue of CKO mice was down-regulated compared to wild type (WT) mice (p < 0.05). The primary alveolar epithelial cells did not express Fut8 protein in CKO mice. CKO mice protein core fucosylation in lung tissues and primary alveolar epithelial cells decreased compared to WT mice. Fut8 enzyme activity in lung tissues of CKO mice was weakener than that one in WT mice (p < 0.05). There is no obvious pathological damage in CKO mice lung tissues. Conclusions: The alveolar epithelial cell-specific Fut8 conditional knockout mouse model was successfully created, which may be helpful to study the role of Fut8 and lung disease.