Molecular determinants of WNT9b responsiveness in nephron progenitor cells

被引:11
作者
Dickinson, Kyle K. [1 ]
Hammond, Leah C. [2 ]
Karner, Courtney M. [3 ]
Hastie, Nicholas D. [4 ]
Carroll, Thomas J. [5 ]
Goodyer, Paul [2 ,6 ]
机构
[1] McGill Univ, Dept Expt Med, Montreal, PQ, Canada
[2] McGill Univ, Dept Human Genet, Montreal, PQ, Canada
[3] Duke Univ, Sch Med, Dept Orthopaed Surg & Cell Biol, Durham, NC USA
[4] Univ Edinburgh, Med Res Council Human Genet Unit, Edinburgh, Midlothian, Scotland
[5] Univ Texas Southwestern, Dept Mol Biol, Dallas, TX USA
[6] McGill Univ, Ctr Hlth, Dept Paediat, Montreal, PQ, Canada
来源
PLOS ONE | 2019年 / 14卷 / 04期
关键词
WILMS-TUMOR SUPPRESSOR; WNT/BETA-CATENIN; SELF-RENEWAL; STEM-CELLS; KIDNEY; MESENCHYME; LGR5; DIFFERENTIATION; MORPHOGENESIS; RECEPTORS;
D O I
10.1371/journal.pone.0215139
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Primed nephron progenitor cells (NPCs) appear in metanephric mesenchyme by E11.5 and differentiate in response to the inductive WNT9b signal from the ureteric bud. However, the NPC WNT-receptor complex is unknown. We obtained M15 cells from E10.5 mesonephric mesenchyme and systematically analyzed components required for canonical WNT9b-responsiveness. When M15 cells were transfected with a beta-catenin luciferase reporter plasmid, exposure to recombinant WNT9b resulted in minimal luciferase activity. We then analyzed mRNA-expression of WNT-pathway components and identified Fzd1-6 and Lrp6 transcripts but not Rspo1. When M15 cells were treated with recombinant RSPO1 the response to transfected WNT9b was augmented 4.8-fold. Co-transfection of M15 cells with Fzd5 (but no other Fzd family member) further increased the WNT9b signal to 16.8-fold and siRNA knockdown of Fzd5 reduced the signal by 52%. Knockdown of Lrp6 resulted in 60% WNT9b signal reduction. We confirmed Fzd5, Lrp6 and Rspo1 mRNA expression in CITED1(+) NPCs from E15.5 embryonic mouse kidney. Thus, while many WNT signaling-pathway components are present by E10.5, optimum responsiveness of E11.5 cap mesenchyme requires that NPCs acquire RSPO1, FZD5 and LRP6.
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页数:16
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