Hox Abdominal-B Genes in the Developing Kidney

The Hox complex consists of 39 genes arranged in 4 clusters of flanking genes and 13 paralogous groups in mammals. To assess the functional redundancy of Hox abdominal-B genes during renal development, we used a modified recombineering strategy to simultaneous introduce frameshift mutations into the Hox9, Hox10, and Hox11 flanking genes of the HoxA, HoxC, and HoxD paralogous groups. We performed RNA seq on whole kidneys at E18.5 in triplicates for representative genotypes including: wild type; Hoxa9,10,11-/- HoxC9,10,11+/-, Hoxa9,10,11+/- HoxC9,10,11-/-, Hoxa9,10,11-/- HoxC9,10,11-/-. Our results suggest that the loss of Hox function results in a partial metanephric to mesonephric transformation, with tubules co-expressing markers of both proximal tubules and collecting ducts, as well as markers of mesonephric-derived epididymis tubules. mRNA profiles were generated by performing RNA-seq on whole kidneys at E18.5 in triplicates for Hox mutant genotypes including: 1) wild type; 2) Hoxa9,10,11-/- HoxC9,10,11+/-, 3) Hoxa9,10,11+/- HoxC9,10,11-/-, and 4) Hoxa9,10,11-/- HoxC9,10,11-/- by deep sequencing using Illumina Hi-Seq 2500哺乳动物的同源框基因簇（Hox complex）包含39个基因，分为4个侧翼基因簇与13个旁系同源组。为探究肾脏发育过程中Hox腹部B型基因的功能冗余性，本研究采用改良的同源重组工程（recombineering）策略，同时对HoxA、HoxC及HoxD旁系同源组的Hox9、Hox10与Hox11侧翼基因引入移码突变。我们针对以下代表性基因型的胚胎发育第18.5天（E18.5）全肾样本，开展了三次生物学重复的RNA测序（RNA-seq）：野生型、Hoxa9、10、11纯合缺失且HoxC9、10、11杂合缺失、Hoxa9、10、11杂合缺失且HoxC9、10、11纯合缺失，以及Hoxa9、10、11纯合缺失且HoxC9、10、11纯合缺失。研究结果显示，Hox功能缺失会引发部分后肾向中肾的转分化，相关肾小管会同时表达近端小管与集合管的标志物，同时还会表达中肾来源的附睾小管标志物。本研究通过Illumina Hi-Seq 2500平台开展深度测序，生成了上述Hox突变基因型的全肾mRNA表达谱，所有样本均为E18.5全肾的三次生物学重复。