4.5 Article

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

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出版社

BIOLIFE SAS
DOI: 10.23812/j.biol.regul.homeost.agents.20223606.213

关键词

core fucosylation; conditional knockout; Fut8; alveolar epithelial cell

资金

  1. National Natural Sci-ence Foundation of China [81900057]

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In this study, a mouse model with lung-specific conditional knockout of Fut8 gene was created, which may be useful for studying the relationship between Fut8 and lung diseases.
Aim: To create a mice model to better evaluate the relationship between alpha 1,6-fucosyltransferase (Fut8) with lung diseases like pul-monary 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 re-action (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 histolog-ical 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.

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