XB-IMG-176432
Xenbase Image ID: 176432
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Fig. 3. The WGA tracer co-localizes with Twist1-expressing cells in the ventral route. (A) Scheme of the experimental procedure: postero-anterior sections of embryos (a to d) at stages 18, 21, 23 and 25 were stained by a myotome marker (12/101 antibody, green), WGA-rodhamine (red), and Hoechst 33258 nucleus marker (blue). WGA is used here to mark all cells and to better visualize the notochord contour. Brackets design somitic ventral migrating cells. Rectangle dotted line designs the area magnification at the bottom (stage 18) or at the right side (stages 21, 23 and 25) of each section (a’ to d’). Scale bars: 100 μm. (B) Postero-anterior expression of Twist1 at stage 18, 21, 23 and 25 on transverse sections (a to d). (C) Scheme of the experimental procedure: lineage tracing experiments with WGA-rhodamine (red) injected in the Meox2 expression domain at stage 13, fixed and sectioned at stage 22/23, then sections were stained with a myotome marker (12/101 antibody, green) and Hoechst 33258 nucleus marker. (D) Scheme of the experimental procedure: lineage tracing experiments with WGA-rhodamine (red) injected in the Meox2 expression domain at stage 13, fixed and sectioned at stage 22/23, then slides were stained with Hoechst 33258 nucleus marker. The same slides were next submitted to in situ hybridization with Twist1 probe to evaluate co-staining of WGA tracer with Twist1 expression. Arrows design cells labelled by WGA-rhodamine. (E), (F) and (G). Ablation of lateral somitic domain prevents sclerotome and hypaxial muscles formation. Scheme of the experimental procedure: ablation experiments of the lateral somitic domain at stage 17. Embryos were fixed and analysed for Twist1 (E), MyhE3 (F) and Mef2c (G) expressions (arrows) and in comparison with 12/101 staining in E. Abl. C, sham-operated embryos. Abl. M, medial domain was removed. Abl. L, lateral domain was removed. For statistical analysis, number of embryos is as follows: same expression, faint decreased, strong decreased. Kruskal-Wallis test and Dunn's multiple comparisons test revealed significant difference between Abl. L with Abl. C or Abl. M, for both Twist1 p < 0.0001, (Abl. C 13, 2, 0; Abl. M 13, 3, 0; Abl. L 0, 2, 13), and Mef2c p < 0.0001, (Abl. C 24, 0, 0; Abl. M 15, 6, 0; Abl. L 3, 0, 16) expressions. Mann-Whitney test revealed significant difference between Abl. L with Abl. C for MyhE3 expression p < 0.0001 (Abl. C 15, 0, 0; Abl. L 0, 5, 10). Image published in: Della Gaspera B et al. (2019) Copyright © 2019. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View |