The intrinsic innervation of the lung is derived from neural crest cells as shown by optical projection tomography in Wnt1-Cre;YFP reporter mice
The intrinsic innervation of the lung is derived from neural crest cells as shown by optical projection tomography in Wnt1-Cre;YFP reporter mice
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DOI：
10.1111/j.1469-7580.2010.01295.x
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发表时间：
2010-12-01
影响因子：
2.4
通讯作者：
Burns, Alan J.
中科院分区：
文献类型：
--
作者：
Freem, Lucy J.;Escot, Sophie;Burns, Alan J.
在胚胎肺内，支配气道平滑肌的固有神经节是正常肺发育和功能所必需的。我们通过将Wnt1-Cre小鼠与R26R-EYFP报告小鼠杂交，获得在所有神经嵴细胞（NCCs）及其衍生物中表达黄色荧光蛋白（YFP）的双转基因小鼠，研究了胚胎小鼠肺内神经嵴来源的固有神经元的发育。除了在冷冻肺切片上使用传统的免疫组织化学，通过结合NCCs的遗传标记和光学投影断层扫描（一种新的成像技术，对胚胎内发育器官的3D检查特别有用），在三维上可视化了肺神经分布的复杂组织。胚胎10.5天，yfp阳性的NCCs从食道区迁移到小鼠肺中。这些细胞随后聚集在肺的支气管和上皮小管周围，通过光学投影断层扫描成像的三维肺重建显示，这些细胞形成了一个广泛的分支网络，与发育中的气道相关。yfp阳性细胞也定植在器官型培养中维持的肺中，并以化学吸引的方式响应原癌基因，在转染（RET）配体期间重排，胶质细胞系来源的神经营养因子（GDNF），这表明RET信号通路参与肺内神经元发育。然而，当检查Ret-/-和Gfr α 1-/-胚胎的肺时，分别缺乏Ret受体和GDNF家族受体α 1 （Gfr α 1）共受体，肺神经支配的程度没有明显差异。我们的研究结果表明，小鼠肺的内在神经元来源于NCCs，尽管与这些细胞的发育有关，但RET信号通路的作用需要进一步研究。
Within the embryonic lung, intrinsic nerve ganglia, which innervate airway smooth muscle, are required for normal lung development and function. We studied the development of neural crest-derived intrinsic neurons within the embryonic mouse lung by crossing Wnt1-Cre mice with R26R-EYFP reporter mice to generate double transgenic mice that express yellow fluorescent protein (YFP) in all neural crest cells (NCCs) and their derivatives. In addition to utilizing conventional immunohistochemistry on frozen lung sections, the complex organization of lung innervation was visualized in three dimensions by combining the genetic labelling of NCCs with optical projection tomography, a novel imaging technique that is particularly useful for the 3D examination of developing organs within embryos. YFP-positive NCCs migrated into the mouse lung from the oesophagus region at embryonic day 10.5. These cells subsequently accumulated around the bronchi and epithelial tubules of the lung and, as shown by 3D lung reconstructions with optical projection tomography imaging, formed an extensive, branching network in association with the developing airways. YFP-positive cells also colonized lung maintained in organotypic culture, and responded in a chemoattractive manner to the proto-oncogene, rearranged during transfection (RET) ligand, glial-cell-line-derived neurotrophic factor (GDNF), suggesting that the RET signalling pathway is involved in neuronal development within the lung. However, when the lungs of Ret-/- and Gfr alpha 1-/- embryos, deficient in the RET receptor and GDNF family receptor alpha 1 (GFR alpha 1) co-receptor respectively, were examined, no major differences in the extent of lung innervation were observed. Our findings demonstrate that intrinsic neurons of the mouse lung are derived from NCCs and that, although implicated in the development of these cells, the role of the RET signalling pathway requires further investigation.