Xu Jingyue, Liu Han, Chai Ok Hee, Lan Yu, Jiang Rulang
Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, United States of America.
Department of Anatomy, Chonbuk National University Medical School and Institute for Medical Sciences, Deokjin-gu, Jeonju 561-756, Republic of Korea.
PLoS One. 2016 Jul 21;11(7):e0159597. doi: 10.1371/journal.pone.0159597. eCollection 2016.
Renal hypoplasia is a common cause of pediatric renal failure and several adult-onset diseases. Recent studies have associated a variant of the OSR1 gene with reduction of newborn kidney size and function in heterozygotes and neonatal lethality with kidney defects in homozygotes. How OSR1 regulates kidney development and nephron endowment is not well understood, however. In this study, by using the recently developed CRISPR genome editing technology, we genetically labeled the endogenous Osr1 protein and show that Osr1 interacts with Wt1 in the developing kidney. Whereas mice heterozygous for either an Osr1 or Wt1 null allele have normal kidneys at birth, most mice heterozygous for both Osr1 and Wt1 exhibit defects in metanephric kidney development, including unilateral or bilateral kidney agenesis or hypoplasia. The developmental defects in the Osr1+/-Wt1+/- mouse embryos were detected as early as E10.5, during specification of the metanephric mesenchyme, with the Osr1+/-Wt1+/- mouse embryos exhibiting significantly reduced Pax2-positive and Six2-positive nephron progenitor cells. Moreover, expression of Gdnf, the major nephrogenic signal for inducing ureteric bud outgrowth, was significantly reduced in the metanephric mesenchyme in Osr1+/-Wt1+/- embryos in comparison with the Osr1+/- or Wt1+/- littermates. By E11.5, as the ureteric buds invade the metanephric mesenchyme and initiate branching morphogenesis, kidney morphogenesis was significantly impaired in the Osr1+/-Wt1+/- embryos in comparison with the Osr1+/- or Wt1+/- embryos. These results indicate that Osr1 and Wt1 act synergistically to regulate nephron endowment by controlling metanephric mesenchyme specification during early nephrogenesis.
肾发育不全是小儿肾衰竭和几种成人发病疾病的常见病因。最近的研究表明,OSR1基因的一个变体与杂合子新生儿肾脏大小和功能的降低以及纯合子中伴有肾脏缺陷的新生儿致死率有关。然而,OSR1如何调节肾脏发育和肾单位数量尚不清楚。在本研究中,我们使用最近开发的CRISPR基因组编辑技术对内源性Osr1蛋白进行基因标记,并表明Osr1在发育中的肾脏中与Wt1相互作用。虽然Osr1或Wt1无效等位基因的杂合子小鼠出生时肾脏正常,但大多数Osr1和Wt1均为杂合子的小鼠在肾后肾发育中表现出缺陷,包括单侧或双侧肾缺如或发育不全。早在E10.5,即在肾后肾间充质特化期间,就检测到了Osr1+/-Wt1+/-小鼠胚胎的发育缺陷,Osr1+/-Wt1+/-小鼠胚胎中Pax2阳性和Six2阳性肾单位祖细胞显著减少。此外,与Osr1+/-或Wt1+/-同窝小鼠相比,Osr1+/-Wt1+/-胚胎的肾后肾间充质中诱导输尿管芽生长的主要肾发生信号Gdnf的表达显著降低。到E11.5时,随着输尿管芽侵入肾后肾间充质并启动分支形态发生,与Osr1+/-或Wt1+/-胚胎相比,Osr1+/-Wt1+/-胚胎的肾脏形态发生明显受损。这些结果表明,Osr1和Wt1协同作用,通过在早期肾发生过程中控制肾后肾间充质特化来调节肾单位数量。