Increased levels of mucins in the cystic fibrosis mouse small intestine, and modulator effects of the Muc1 mucin expression
Increased levels of mucins in the cystic fibrosis mouse small intestine, and modulator effects of the Muc1 mucin expression
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DOI：
10.1152/ajpgi.00491.2005
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发表时间：
2006-08-01
影响因子：
4.5
通讯作者：
Hansson, Gunnar C.
中科院分区：
文献类型：
--
作者：
Malmberg, Emily K.;Noaksson, Karin A.;Hansson, Gunnar C.
囊性纤维化(CF)的小鼠模型(CFTR(Tm1UNC)/CFTR(Tm1UNC))显示粘液堆积，并由于剪接改变(Hinojosa-Kurtzberg AM、Johansson MEV、Madsen CS、Hansson GC和Gendler SJ)而增加MUC1粘蛋白mRNA水平。Am J Physiol Gasterintest肝脏生理学284：G853-G862,2003)。然而，目前尚不清楚MUC1是否是导致粘液增加的主要粘蛋白，以及为什么CF/MUC1(-/-)小鼠表现出较低的粘液积累。为了解决这一问题，我们采用氯化胍提取、超速离心和凝胶过滤的方法从CF小鼠的小肠中纯化了粘蛋白，并通过狭缝印迹、凝胶电泳、蛋白质组学和免疫印迹等方法对其进行了分析。携带野生型(WT)MUC1或MUC1(-/-)的正常和CF小鼠，或转人MUC1(MUC1.Tg，在MUC1(-/-)背景下)被分析。与非CF组相比，CF组小鼠体内可溶于和不溶于瓜尼丁的粘蛋白总量增加了10倍，而缺乏MUC1的CF组小鼠的粘蛋白总量介于CF组和非CF组小鼠之间。然而，Cf/MUC1(-/-)小鼠的Muc3(人MUC17的同源基因)水平高于Cf/MUC1小鼠。转基因动物。在CF小鼠中，MUC1粘蛋白的数量增加了几个数量级，但MUC1仍不是主要的粘蛋白。CF组小鼠的大肠不溶性粘液量也增加，这一效应在CF/MUC1(-/-)组小鼠中部分恢复。MUC1(-/-)小鼠结肠粘液层厚度明显低于WT小鼠。结果表明，MUC1不是CF小鼠累积粘液中的主要成分，MUC1可以以一种未知的方式影响其他粘蛋白的数量。
The mouse model (Cftr(tm1UNC)/Cftr(tm1UNC)) for cystic fibrosis (CF) shows mucus accumulation and increased Muc1 mucin mRNA levels due to altered splicing (Hinojosa-Kurtzberg AM, Johansson MEV, Madsen CS, Hansson GC, and Gendler SJ. Am J Physiol Gastrointest Liver Physiol 284: G853-G862, 2003). However, it is not known whether Muc1 is a major mucin contributing to the increased mucus and why CF/Muc1(-/-) mice show lower mucus accumulation. To address this, we have purified mucins from the small intestine of CF mice using guanidinium chloride extraction, ultracentrifugation, and gel filtration and analyzed them by slot blot, gel electrophoresis, proteomics, and immunoblotting. Normal and CF mice with wild-type (WT) Muc1 or Muc1(-/-) or that are transgenic for human MUC1 (MUC1. Tg, on a Muc1(-/-) background) were analyzed. The total amount of mucins, both soluble and insoluble in guanidinium chloride, increased up to 10-fold in the CF mice compared with non-CF animals, whereas the CF mice lacking Muc1 showed intermediate levels between the CF and non-CF mice. However, the levels of Muc3 (orthologue of human MUC17) were increased in the CF/Muc1(-/-) mice compared with the CF/MUC1. Tg animals. The amount of MUC1 mucin was increased several magnitudes in the CF mice, but MUC1 did still not appear to be a major mucin. The amount of insoluble mucus of the large intestine was also increased in the CF mice, an effect that was partially restored in the CF/Muc1(-/-) mice. The thickness of the firmly adherent mucus layer of colon in the Muc1(-/-) mice was significantly lower than that of WT mice. The results suggest that MUC1 is not a major component in the accumulated mucus of CF mice and that MUC1 can influence the amount of other mucins in a still unknown way.