BMP4 promotes development and patterning of the tracheal cartilage and trachealis muscle downstream of epithelial-induced Wnt signaling

By differentially regulating expression of direct Wnt/b-catenin target genes, Bmp4 modulates levels of Wnt signaling necessary to promote cartilaginous identity and progress tracheal chondrogenesis. E13.5, Wls flox/flox, ShhCre+ trachea and e13.5 control tracheas were isolated from mouse embryos and submitted for RNA sequencing. Resulting data was combined with previously generated data to determine gene changes. E13.5, Chondroblasts were isolated by FACS negative selection from, Bmp4flox/flox, gammaSMAeGFP and Bmp4flox/flox, Foxg1Cre+, gammaSMAeGFP mouse trachea using APC-Epcam and GFP to remove epithelial and muscle cells respectively.骨形态发生蛋白4（Bone morphogenetic protein 4, BMP4）通过差异调控Wnt/β-连环蛋白（Wnt/β-catenin）直接靶基因的表达，调控促进软骨细胞表型形成并推进气管软骨发生所需的Wnt信号通路水平。本研究从小鼠胚胎中分离得到胚胎发育第13.5天（E13.5）的Wls flox/flox且Shh-Cre阳性（ShhCre+）气管，以及同阶段对照气管，并对其进行RNA测序；将所得测序数据与既往已生成的数据集合并，以明确基因表达变化情况。此外，在E13.5阶段，研究人员通过荧光激活细胞分选（Fluorescence-activated cell sorting, FACS）阴性筛选法，分别利用APC-EpCAM与绿色荧光蛋白（GFP）去除上皮细胞与肌细胞，从Bmp4 flox/flox、γ-平滑肌肌动蛋白-增强绿色荧光蛋白（gammaSMAeGFP）小鼠气管，以及Bmp4 flox/flox、Foxg1-Cre阳性（Foxg1Cre+）且gammaSMAeGFP小鼠气管中分离得到成软骨细胞（Chondroblasts）。