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. 2010 Nov 17;5(11):e15471. doi: 10.1371/journal.pone.0015471

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Croizier, 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 properly credited.

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(A) Histogram summarizing precedent findings in the rat concerning the genesis of MCH, MCH/CART/NK3 and Hcrt neurons in the hypothalamus [12],[18]. (B–F) A similar study combining BrdU, immunohistochemistry and in situ hybridization was conducted in mouse to compare the birthdates of MCH and Hcrt neurons. (B) Photomicrograph illustrating a neuron double-labeled for BrdU (red immunohistochemistry) and MCH (in situ hybridization). (C) Dual immunofluorescence to identify Hcrt (green) and BrdU in the same neurons. Only neurons displaying a densely BrdU-labeled nucleus were taken into consideration. (D–F) Histograms of MCH/BrdU, NK3/BrdU and Hcrt/BrdU neurons. MCH/BrdU positive neurons are seen from E9 to E14, with a peak at E10 (D). NK3/BrdU positive cells in the zona incerta/LHA were observed only at E11 and E12 (E). On the same material, double labeled Hcrt/BrdU perikarya were observed mainly at E10 (F). LHA: lateral hypothalamic area.