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. 2019 Mar 14;12:62. doi: 10.3389/fnmol.2019.00062

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Copyright © 2019 Wever, Largo-Barrientos, Hoekstra and Smidt.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Lmx1b represses Ahd2 independent of Pitx3. The expression patterns of Adh2, En1 and Th were examined using in situ hybridization and the mRNA levels for Lmx1b, Ahd2, En1, Cck and Th were verified using qPCR. (A) Pitx3Cre driven loss of Lmx1b in Pitx3CreCre animals caused an increase in Ahd2 expression in the medial sections of the E14.5 midbrain (black arrowheads). (B,C) The expression pattern of En1 did not show clear differences between Pitx3CreCre; Lmx1b +/+ and the Pitx3CreCre Lmx1b mutant and neither did Th. (D) The mRNA expression levels of Lmx1b were down-regulated with ~76% in conditional mutant (gray bar, n = 3, *p < 0.05, one-tailed), while Ahd2 shows an increase of ~160% in the Pitx3CreCre; Lmx1b L/L (gray bar, n = 3, *p < 0.05, one-tailed) compared to Pitx3CreCre; Lmx1b +/+ littermates (red line, n = 4). The mRNA expression levels of En1, Cck and Th did not show significant changes. Wildtype was set at 1. Scale bar = 100 μM.