School of Biological Sciences, University of Manchester, Manchester, M13 9PT, UK.
Lydia Becker Institute of Immunology and Inflammation, University of Manchester, Manchester, M13 9PT, UK.
Sci Rep. 2024 Oct 26;14(1):25555. doi: 10.1038/s41598-024-76482-y.
The TMEM16A chloride channel is proposed as a therapeutic target in cystic fibrosis, where activation of this ion channel might restore airway surface hydration and mitigate respiratory symptoms. While TMEM16A is associated with increased mucin production under stimulated or pro-inflammatory conditions, its role in baseline mucin production, secretion and/or maturation is less well understood. Here, we use the Xenopus tadpole skin mucociliary surface as a model of human upper airway epithelium to study Tmem16a function in mucus production. We found that Xenopus tropicalis Tmem16a is present at the apical membrane surface of tadpole skin small secretory cells that express canonical markers of mammalian "goblet cells" such as Foxa1 and spdef. X. tropicalis Tmem16a functions as a voltage-gated, calcium-activated chloride channel when transfected into mammalian cells in culture. Depletion of Tmem16a from the tadpole skin results in dysregulated mucin maturation post-secretion, with secreted mucins having a disrupted molecular size distribution and altered morphology assessed by sucrose gradient centrifugation and electron microscopy, respectively. Our results show that in the Xenopus tadpole skin, Tmem16a is necessary for normal mucus barrier formation and demonstrate the utility of this model system to discover new biology relevant to human mucosal biology in health and disease.
TMEM16A 氯离子通道被提议作为囊性纤维化的治疗靶点,激活该离子通道可能恢复气道表面的水合作用并减轻呼吸道症状。虽然 TMEM16A 在受刺激或促炎条件下与增加粘蛋白产生有关,但它在基线粘蛋白产生、分泌和/或成熟中的作用了解较少。在这里,我们使用非洲爪蟾蝌蚪皮肤粘膜表面作为人类上呼吸道上皮的模型来研究 Tmem16a 在粘液产生中的作用。我们发现,非洲爪蟾 Tmem16a 存在于表达哺乳动物“杯状细胞”典型标志物(如 Foxa1 和 spdef)的蝌蚪皮肤小分泌细胞的顶膜表面。X. 热带 Tmem16a 在培养的哺乳动物细胞中转染后作为电压门控、钙激活氯离子通道发挥作用。从蝌蚪皮肤中耗尽 Tmem16a 会导致分泌后粘蛋白成熟失调,通过蔗糖梯度离心和电子显微镜分别评估,分泌的粘蛋白的分子大小分布中断,形态改变。我们的结果表明,在非洲爪蟾蝌蚪皮肤中,Tmem16a 对于正常的粘液屏障形成是必要的,并证明了该模型系统在发现与健康和疾病中的人类粘膜生物学相关的新生物学方面的实用性。