[1]包和婧,马树东.Shh信号通路对小鼠胚胎肺发育的调控作用[J].南方医科大学学报,2018,(03):274.
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Shh信号通路对小鼠胚胎肺发育的调控作用()
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《南方医科大学学报》[ISSN:1673-4254/CN:44-1627/R]

卷:
期数:
2018年03期
页码:
274
栏目:
出版日期:
2018-04-03

文章信息/Info

Title:
Regulatory role of Shh signaling pathway in lung development in fetal mice
作者:
包和婧马树东
关键词:
Shh信号通路基因敲除肺发育免疫荧光
Keywords:
Shh signaling pathway gene knockdown lung development immunofluo- rescence
摘要:
目的探讨经典Shh信号通路对小鼠胚胎肺发育过程中上皮、间质(支气管软骨、平滑肌)发育的调控作用。方法利用免 疫组化检测Shh通路受体Smo(Smoothened)以及血小板源性生长因子受体(Pdgfr-α)的表达;利用Pdgfr-α间质特异性表达的特 点构建Pdgfr-α-cre,通过他莫昔芬的诱导,在E12.5-E16.5(E, embryo)天转基因小鼠肺间质中特异性敲除Shh 关键信号分子 Smo;利用免疫荧光观察E12.5-E16.5小鼠胚胎肺间质特异性敲除Shh信号通路之后,小鼠胚胎肺发育过程中上皮、间质(支气管 软骨、平滑肌)发育的改变,探讨Shh信号通路在小鼠胚胎肺发育过程中对上皮-间质转化的作用。结果Smo在假腺期早期的肺 上皮与间质中显著表达,后表达量逐渐下调,主要集中在肺间质,Pdgfr-α在假腺期早期胚胎肺中集中在远端肺上皮及间质,后期 逐渐往近端间质发展,直至集中在主支气管周围的近端间质;首次利用Pdgfr-α-Cre系统在间质中特异性敲除Smo,成功制备了 Smo缺失小鼠模型;与对照组肺相比,E12.5-E16.5基因敲除组小鼠的肺体积缩小,支气管分支形成减少;近端上皮指标P63的表 达量下降(P<0.05);平滑肌标志物表达水平发生改变;支气管软骨发育滞后,黏蛋白表达量下降。结论Shh信号通路的时空特 异表达在小鼠胚胎肺上皮、间质(支气管软骨、平滑肌)发育的起着重要的调控作用。
Abstract:
Objective To investigate the regulatory role of classical Shh signaling pathway in the development of the epithelium and mesenchyme (bronchial cartilage and smooth muscles) during lung development in fetal mice. Methods Immunohistochemical technique was used to detect the expression of Shh signaling pathway receptor Smo and Pdgfr-α in murine fetal lungs to explore the spatial and temporal characteristics of their expression. Based on the interstitial specificity of Pdgfr-α expression, we constructed a Pdgfr-α-cre to establish a E12.5 - E16.5 transgenic mice with specific knockout of the key Shh signaling molecule Smo in the pulmonary interstitium with tamoxifen induction. Immunofluorescence technique was used to observe the epithelium and mesenchyme (bronchial cartilage and smooth muscle) during fetal lung development in the transgenic mice to assess the role of Shh signaling pathway in the epithelial-to-mesenchymal (EMT) transition during the lung development. Results Smo was highly expressed in the epithelial and stromal lung tissues in the pseudoglandular stage and was gradually lowered over time with its distribution mainly in the interstitial tissues. Pdgfr-α was enriched in the distal lung epithelial and mesenchy tissues in early embryonic lungs and gradually migrated to the proximal stroma until becoming concentrated around the main bronchial proximal stroma. We successfully specific established mouse models of specific mesenchymal Smo knockout. Compared with the control group, the transgenic mice during E12.5-E16.5 showed significantly reduced lung the volume and bronchial branching with also decreased expression of the proximal epithelial P63 (P<0.05). The transgenic mice exhibited alterations in the expression of α-smooth muscle actin with delayed bronchial cartilage development and decreased expression of mucoprotein. Conclusion The temporospatial specific expression of Shh signaling pathway plays an important role in developmental regulation of mouse embryonic lung epithelium and mesenchyme (bronchial cartilage and smooth muscle).

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更新日期/Last Update: 1900-01-01