XB-IMG-84157
Xenbase Image ID: 84157
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Figure 4.
miR-142-3pActsattheTopoftheRegulatoryHierarchyControllingSpecificationofHSCandEndothelialCellPrecursorsintheDLP
(A) A provisional GRN controlling hemangioblast specification (Ciau-Uitz et al., 2010, 2013; Liu et al., 2008; Walmsley et al., 2002).
(B and C) Initiation of the hemangioblast program is defective in miR-142-3p MO embryos. (B) WISH showing that expression of early regulators, fli1, flk1, and etv2, is already highly reduced or absent in the DLP (arrows) in miR-142-3p morphants at stage 22 (initiation phase). (C) WISH analysis showing that the expression of hematopoietic gata2 and scl) and endothelial (fli1 and etv2) TFs in the DLP (arrows), are all dependent on miR-142-3p at stages 25–26. Boxed region of the fli1 panels is magnified to show that the expression of fli1 in the pronephric duct (arrowheads) abutting the DLP is unaffected in the morpants, whereas the expression of fli1 in the DLP (white dashed outline and arrows) is abrogated by the miR-142-3p MO. Formation of endothelial cells in the VBI is also disrupted by the MO as shown by reduced etv2 staining (arrowhead).
(D and E) Hemangioblast specification is normal in embryos injected with the control mismatch MO and miR-142-5p MO showing that the effect is specific to the miR-142-3p MO. (D) WISH analysis of etv2 and scl expression in the DLP (arrows) of control mismatch morphants and (E) gata2 and etv2 expression in the DLP (arrows) of miR-142-5p morphants.
(F) Formation of primitive blood is defective in miR-142-3p morphants. Top: Erythroid differentiation is absent from the pVBI region as shown by WISH analysis of aT4-globin expression. Bottom: reduction in formation of myeloid cells as shown by WISH analysis of elas1 expression.
The number of embryos represented by each panel, out of the number analyzed, is indicated in the top right corner. The anterior of the embryo is to the left and dorsal to the top in all images.
See also Figure S3. Image published in: Nimmo R et al. (2013) Copyright © 2013. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View |