LIRIC UMR 995, Univ. Lille, Inserm, CHU Lille, F-59000 Lille, France.
Sci Rep. 2017 Mar 15;7:44591. doi: 10.1038/srep44591.
Gel-forming mucins are macromolecules produced by goblet cells and responsible for the mucus gel formation. Changes in goblet cell density and in gel-forming mucin production have emerged as sensitive indicators for mucosal diseases. A Muc5b-GFP tagged reporter mouse was used to assess Muc5b production in mouse tissues by immunofluorescence microscopy and fluorescent activity using stereromicroscopy and probe-based confocal laser endomicroscopy. Muc5b production was followed longitudinally by recording the fluorescent activity in vagina and in embryonic lung explants under stimulation by interleukin 13. We show that the GFP is easily visualized in the mouse adult ear, nose, trachea, gallbladder, and cervix. Live Muc5b is also easily monitored in the nasal cavity, trachea and vagina where its production varies during the estrus cycle with a peak at the proestrus phase and in pregnant mice. Explant culture of reporter mouse embryonic whole lung shows that interleukin 13 stimulates Muc5b production. The transgenic Muc5b-GFP mouse is unique and suitable to study the mechanisms that regulate Muc5b production/secretion and mucous cell differentiation by live imaging and can be applied to test drug efficacy in mucosal disease models.
胶状黏液蛋白是由杯状细胞产生的大分子物质，负责黏液凝胶的形成。杯状细胞密度和胶状黏液蛋白产生的变化已成为黏膜疾病的敏感指标。通过免疫荧光显微镜和立体显微镜以及基于探针的共聚焦激光内窥检查术，使用 Muc5b-GFP 标记的报告小鼠来评估小鼠组织中的 Muc5b 产生。通过记录白细胞介素 13 刺激下阴道和胚胎肺外植体中的荧光活性，对 Muc5b 进行纵向跟踪。我们发现 GFP 在成年小鼠的耳朵、鼻子、气管、胆囊和子宫颈中很容易被观察到。活的 Muc5b 也很容易在鼻腔、气管和阴道中被监测到，其在发情周期中变化，在发情前期达到峰值，在怀孕的小鼠中也会增加。报告小鼠胚胎全肺外植体培养表明白细胞介素 13 刺激 Muc5b 的产生。这种转基因 Muc5b-GFP 小鼠是独特的，适合通过活体成像研究调节 Muc5b 产生/分泌和黏液细胞分化的机制，并可应用于测试黏膜疾病模型中的药物疗效。
