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利用一种新的体内模型研究黏液纤毛上皮发育,以鉴定新的纤毛发生因子。

Identification of novel ciliogenesis factors using a new in vivo model for mucociliary epithelial development.

作者信息

Hayes Julie M, Kim Su Kyoung, Abitua Philip B, Park Tae Joo, Herrington Emily R, Kitayama Atsushi, Grow Matthew W, Ueno Naoto, Wallingford John B

机构信息

Department of Molecular Cell and Developmental Biology, and Institute for Cellular and Molecular Biology, University Station C1000, University of Texas, Austin, TX 78712, USA.

出版信息

Dev Biol. 2007 Dec 1;312(1):115-30. doi: 10.1016/j.ydbio.2007.09.031. Epub 2007 Sep 26.

Abstract

Mucociliary epithelia are essential for homeostasis of many organs and consist of mucus-secreting goblet cells and ciliated cells. Here, we present the ciliated epidermis of Xenopus embryos as a facile model system for in vivo molecular studies of mucociliary epithelial development. Using an in situ hybridization-based approach, we identified numerous genes expressed differentially in mucus-secreting cells or in ciliated cells. Focusing on genes expressed in ciliated cells, we have identified new candidate ciliogenesis factors, including several not present in the current ciliome. We find that TTC25-GFP is localized to the base of cilia and to ciliary axonemes, and disruption of TTC25 function disrupts ciliogenesis. Mig12-GFP localizes very strongly to the base of cilia and confocal imaging of this construct allows for simple visualization of the planar polarity of basal bodies that underlies polarized ciliary beating. Knockdown of Mig12 disrupts ciliogenesis. Finally, we show that ciliogenesis factors identified in the Xenopus epidermis are required in the midline to facilitate neural tube closure. These results provide further evidence of a requirement for cilia in neural tube morphogenesis and suggest that genes identified in the Xenopus epidermis play broad roles in ciliogenesis. The suites of genes identified here will provide a foundation for future studies, and may also contribute to our understanding of pathological changes in mucociliary epithelia that accompany diseases such as asthma.

摘要

黏液纤毛上皮对许多器官的内环境稳定至关重要,由分泌黏液的杯状细胞和纤毛细胞组成。在此,我们展示了非洲爪蟾胚胎的纤毛表皮,作为黏液纤毛上皮发育体内分子研究的便捷模型系统。使用基于原位杂交的方法,我们鉴定出许多在分泌黏液的细胞或纤毛细胞中差异表达的基因。聚焦于在纤毛细胞中表达的基因,我们鉴定出了新的纤毛发生候选因子,包括一些当前纤毛组中不存在的因子。我们发现TTC25 - GFP定位于纤毛基部和纤毛轴丝,TTC25功能的破坏会干扰纤毛发生。Mig12 - GFP非常强烈地定位于纤毛基部,对该构建体的共聚焦成像允许简单地可视化构成极化纤毛摆动基础的基体平面极性。Mig12的敲低会干扰纤毛发生。最后,我们表明在非洲爪蟾表皮中鉴定出的纤毛发生因子在中线是神经管闭合所必需的。这些结果为神经管形态发生中对纤毛的需求提供了进一步证据,并表明在非洲爪蟾表皮中鉴定出的基因在纤毛发生中发挥广泛作用。这里鉴定出的基因组合将为未来研究提供基础,也可能有助于我们理解伴随哮喘等疾病的黏液纤毛上皮的病理变化。

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