@article{805362a2c32943fda73c6acbcb7e99c1,
title = "Molecular determinants of WNT9b responsiveness in nephron progenitor cells",
abstract = "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 β-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.",
author = "Dickinson, {Kyle K.} and Hammond, {Leah C.} and Karner, {Courtney M.} and Hastie, {Nicholas D.} and Carroll, {Thomas J.} and Paul Goodyer",
note = "Funding Information: This work was supported by operating grants to P.G from the Kidney Foundation of Canada (2501 – https://www.kidney.ca), the Canadian Institutes of Health/ Fonds de recherche du Qu{\'e}bec - Sant{\'e}/ERA-Net for Research Programs on Rare Diseases (6221 and 7628 – http://www.cihr-irsc.gc.ca/e/51266.html) who also gave advice and critical analysis of the manuscript. P.G also received an operating grant from the Research Institute of the McGill University Health Center (6237 – http://rimuhc.ca). K.D was the recipient of a graduate studentship award from the Research Institute of McGill University Health Centre and Desjardins Group. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors would like to acknowledge background information drawn from the GUDMAP consortium public database (www.GUDMAP.org). We would like to acknowledge co-investigators of the CIHR/FRSQ/ERARE consortium who gave advice and critical analysis of the manuscript. Publisher Copyright: {\textcopyright} 2019 Dickinson et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.",
year = "2019",
month = apr,
doi = "10.1371/journal.pone.0215139",
language = "English (US)",
volume = "14",
journal = "PloS one",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "4",
}