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Retinoic acid control of pax8 during renal specification of Xenopus pronephros involves hox and meis3

Durant-Vesga Jennifer, Suzuki Nanoka, View ORCID ProfileOchi Haruki, View ORCID ProfileLe Bouffant Ronan, Eschstruth Alexis, View ORCID ProfileOgino Hajime, View ORCID ProfileUmbhauer Muriel, View ORCID ProfileRiou Jean-François
doi: https://doi.org/10.1101/2022.06.21.496994
Durant-Vesga Jennifer
1Sorbonne Université, CNRS, Institut de Biologie Paris-Seine, IBPS, Laboratoire de Biologie du Développement, UMR7622, 9, quai Saint-Bernard, 75252 Paris cedex05, France
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Suzuki Nanoka
2Institute for Promotion of Medical Science Research, Faculty of Medicine, Yamagata University, 2-2-2 Iida-Nishi, Yamagata 990-9585, Japan
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Ochi Haruki
2Institute for Promotion of Medical Science Research, Faculty of Medicine, Yamagata University, 2-2-2 Iida-Nishi, Yamagata 990-9585, Japan
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Le Bouffant Ronan
1Sorbonne Université, CNRS, Institut de Biologie Paris-Seine, IBPS, Laboratoire de Biologie du Développement, UMR7622, 9, quai Saint-Bernard, 75252 Paris cedex05, France
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Eschstruth Alexis
1Sorbonne Université, CNRS, Institut de Biologie Paris-Seine, IBPS, Laboratoire de Biologie du Développement, UMR7622, 9, quai Saint-Bernard, 75252 Paris cedex05, France
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Ogino Hajime
3Amphibian Research Center / Graduate School of Integrated Sciences for Life, Hiroshima University, 1-3-1 Kagami-yama, Higashi-Hiroshima, Hiroshima 739-8526, Japan
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Umbhauer Muriel
1Sorbonne Université, CNRS, Institut de Biologie Paris-Seine, IBPS, Laboratoire de Biologie du Développement, UMR7622, 9, quai Saint-Bernard, 75252 Paris cedex05, France
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Riou Jean-François
1Sorbonne Université, CNRS, Institut de Biologie Paris-Seine, IBPS, Laboratoire de Biologie du Développement, UMR7622, 9, quai Saint-Bernard, 75252 Paris cedex05, France
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  • For correspondence: jean-francois.riou@sorbonne-universite.fr
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Abstract

Development of the Xenopus pronephros relies on renal precursors grouped at neurula stage into a specific region of dorso-lateral mesoderm called the kidney field. Formation of the kidney field at early neurula stage is dependent on retinoic (RA) signaling acting upstream of renal master transcriptional regulators such as pax8 or lhx1. Although lhx1 might be a direct target of RA-mediated transcriptional activation in the kidney field, how RA controls the emergence of the kidney field remains poorly understood. In order to better understand RA control of renal specification of the kidney field, we have performed a transcriptomic profiling of genes affected by RA disruption in lateral mesoderm explants isolated prior to the emergence of the kidney field and cultured at different time points until early neurula stage. Besides genes directly involved in pronephric development (pax8, lhx1, osr2, mecom), hox (hoxa1, a3, b3, b4, c5 and d1) and the hox co-factor meis3 appear as a prominent group of genes encoding transcription factors (TFs) downstream of RA. Supporting the idea of a role of meis3 in the kidney field, we have observed that meis3 depletion results in a severe inhibition of pax8 expression in the kidney field. Meis3 depletion only marginally affects expression of lhx1 and aldh1a2 suggesting that meis3 principally acts upstream of pax8. Further arguing for a role of meis3 and hox in the control of pax8, expression of a combination of meis3, hoxb4 and pbx1 in animal caps induces pax8 expression, but not that of lhx1. The same combination of TFs is also able to transactivate a previously identified pax8 enhancer, Pax8-CNS1. Mutagenesis of potential PBX-Hox binding motifs present in Pax8-CNS1 further allows to identify two of them that are necessary for transactivation. Finally, we have tested deletions of regulatory sequences in reporter assays with a previously characterized transgene encompassing 36.5 kb of the X. tropicalis pax8 gene that allows expression of a truncated pax8-GFP fusion protein recapitulating endogenous pax8 expression. This transgene includes three conserved pax8 enhancers, Pax8-CNS1, Pax8-CNS2 and Pax8-CNS3. Deletion of Pax8-CNS1 alone does not affect reporter expression, but deletion of a 3.5kb region encompassing Pax8-CNS1 and Pax8-CNS2 results in a severe inhibition of reporter expression both in the otic placode and kidney field domains.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted June 21, 2022.
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Retinoic acid control of pax8 during renal specification of Xenopus pronephros involves hox and meis3
Durant-Vesga Jennifer, Suzuki Nanoka, Ochi Haruki, Le Bouffant Ronan, Eschstruth Alexis, Ogino Hajime, Umbhauer Muriel, Riou Jean-François
bioRxiv 2022.06.21.496994; doi: https://doi.org/10.1101/2022.06.21.496994
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Retinoic acid control of pax8 during renal specification of Xenopus pronephros involves hox and meis3
Durant-Vesga Jennifer, Suzuki Nanoka, Ochi Haruki, Le Bouffant Ronan, Eschstruth Alexis, Ogino Hajime, Umbhauer Muriel, Riou Jean-François
bioRxiv 2022.06.21.496994; doi: https://doi.org/10.1101/2022.06.21.496994

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