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. 2017 May 26;10:157. doi: 10.3389/fnmol.2017.00157

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Copyright © 2017 Kabayiza, Masgutova, Harris, Rucchin, Jacob and Clotman.

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) or licensor 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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OC factors control dI3 distribution. (A–M) Differentiation and distribution of dI3 interneurons in control or Hnf6/Oc2-/- embryos. At e10.5 (A,B) and e11.5 (C,D), dI3 interneurons containing Isl1 and Tlx3 in Hnf6/Oc2-/- embryos (B,D) are detected in numbers and distributions comparable to those observed in control embryos (A,C). Tlx3+Isl1- cells next to the ventricular zone in e11.5 control embryo are late-born dILB (C). (E–F) Quantitative analysis of control or Hnf6/Oc2-/- embryos at e10.5 (E, n = 4) and e11.5 (F, n = 4) shows no statistically significant difference in the number of dI3 cells. At e12.5, Isl1 combined with Tlx3, Brn3a or Arx identifies dI3 interneurons in the central intermediate spinal cord of control (H–J) or Hnf6/Oc2-/- (K–M) embryos. The number of dI3 cells is comparable in control and mutant embryos, and Tlx3, Brn3a and Arx are detected in the Isl1-positive cells (insets in H–M) indicating that the differentiation of dI3 interneurons is not affected by the absence of Onecut factors. Quantitative analysis shows similar numbers of dI3 cells in control or mutants embryos (G, control n = 6, mutant n = 7). However, the distribution of these cells seems different in Hnf6/Oc2-/- embryos (arrows). (N–Q) Density plots of dI3 interneuron distribution at brachial (N,P) or thoracic (O,Q) levels in control (N,O) or Hnf6/Oc2-/- mutant (P,Q) spinal cord at e12.5 show an alteration in the localization of dI3 cells in mutant embryos. The dI3 interneurons migrate as a more compact population in a general dorso-medial to ventro-medial direction, with the densely-packed cells closer to the migration front. Analysis was performed on at least three sections from five control and six mutant embryos (n > 2600 cells per condition; p ≤ 0.001). The pictures show left hemisections. Dashed lines delineate the spinal cord and the lumen of the ventricle. Insets are magnified views of boxed regions and show Tlx3, Brn3a and Arx labeling, respectively. Arrowheads point to properly-located dI3 neurons. Arrows indicate unproperly-located dI3 neurons in the Hnf6/Oc2-/- mutant embryos. Mean values ±SEM. Scale bars = 100 μm.