鼠胚呼吸内胚层相关第二生心区发育中的上皮-间充质转化

师亮 李慧超 景雅* 谢建山 陈浩 蔡玉瑾 杨艳萍

doi:10.16098/j.issn.0529-1356.2018.04.011

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解剖学报 ›› 2018, Vol. 49 ›› Issue (4) : 480-485. DOI: 10.16098/j.issn.0529-1356.2018.04.011

组织学胚胎学发育生物学

鼠胚呼吸内胚层相关第二生心区发育中的上皮-间充质转化

师亮¹李慧超^1,2 景雅^1* 谢建山¹ 陈浩^1,3 蔡玉瑾¹ 杨艳萍¹

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Epithelial-mesenchymal transition during the development of pulmonary endoderm-associated second heart field in mouse embryos

SHI Liang¹ LI Hui-chao ^1,2 JING Ya ^1* XIE Jian-shan¹ CHEN Hao^1,3 CAI Yu-jin¹YANG Yan-ping¹

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摘要

目的探讨上皮-间充质转化与鼠胚呼吸内胚层相关第二生心区发育的关系。方法用免疫印迹法检测转化生长因子β2（TGF-β2）和转化生长因子β3（TGF-β3）在胚龄9～14 d小鼠胚胎心脏的表达情况。另用叉头框蛋白A2（Foxa2）、胰岛因子1（ISL-1）、层黏连蛋白（LN）及上皮型钙黏蛋白（E-cad）抗体，对35例胚龄10～12 d小鼠胚胎心连续石蜡切片进行免疫组织化学或免疫荧光染色。结果 TGF-β2和TGF-β3蛋白于胚龄10～12 d在小鼠胚胎心脏均有表达。在上述呼吸内胚层相关第二生心区发育的时间点，均可见ISL-1阳性呼吸内胚层与周围ISL-1阳性第二生心区间充质分界不清或混为一体的现象，同时伴有相应部位基底膜的断续状或者模糊不清状改变；尤其在胚龄11 d和12 d，部分呼吸内胚层或其实心细胞索细胞失去Foxa2表达，呈较强ISL-1表达，同时伴有基底膜的断裂或者缺失，E-cad表达减弱，细胞间隙出现，甚至从内胚层脱离，失去上皮细胞的顶底极性。结论 TGF-β信号调节的上皮-间充质转化参与鼠胚呼吸内胚层相关第二生心区发育形成。

Abstract

Objective To explore the relationship between epithelial-mesenchymal transition (EMT) andthe development of pulmonary endoderm-associated second heart field in mouse embryos. Methods The transforming growth factor-β(TGF-β) 2 and TGF-β3 expressions in mouse embryonic hearts from embryonic day (ED) 9 to ED14 were detected by Western blotting. Both the immunohistochemical and immunofluorescent staining method were used to reveal forkhead box A2 (Foxa2), islet-1 (ISL-1), laminin (LN) and E-cadherin (E-cad) expressions in serial sections of 35 mouse embryos from ED10 to ED12. Results From ED10 to ED12, both TGF-β2 and TGF-β3 expressed in mouse embryonic heart. In the pulmonary endoderm-associated second heart field development between ED10 and ED12, there was no clear boundary between some ISL-1 positive pulmonary endoderm and surrounding ISL-1 positive second heart field mesenchym which seemed to be intermingled. At the same time, the basement membrane underlying the pulmonary endoderm became interrupted or blurred. Especially at ED11 and ED12, some pulmonary endoderm or its solid cord cells lost Foxa2 expression and showed strongly ISL-1 positive, accompanied by their basement membrane interrupting or missing, E-cad expression decreasing, obvious intercellular space emerging and detachment from endoderm, which suggested the loss of apico-basal polarity. Conclusion Epithelial-mesenchymal transition is involved in the development of pulmonary endoderm-associated second heart field of mouse embryos.

关键词

上皮-间充质转化 / 呼吸内胚层 / 第二生心区 / 转化生长因子β / 免疫荧光 / 免疫印迹法 / 小鼠

Key words

Epithelial-mesenchymal transition

/ Pulmonary endoderm / Second heart field / Transforming growth factor-β / Immunofluorescence / Western blotting / Mouse

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师亮李慧超景雅谢建山陈浩蔡玉瑾杨艳萍. 鼠胚呼吸内胚层相关第二生心区发育中的上皮-间充质转化[J]. 解剖学报. 2018, 49(4): 480-485 https://doi.org/10.16098/j.issn.0529-1356.2018.04.011

SHI Liang LI Hui-chao JING Ya XIE Jian-shan CHEN Hao CAI Yu-jin YANG Yan-ping. Epithelial-mesenchymal transition during the development of pulmonary endoderm-associated second heart field in mouse embryos[J]. Acta Anatomica Sinica. 2018, 49(4): 480-485 https://doi.org/10.16098/j.issn.0529-1356.2018.04.011

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