XB-IMG-81022
Xenbase Image ID: 81022
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Fig. 5. Overexpression of Osr genes promotes ectopic kidney development. (A–L) Lateral views of stage 25 (A–F), stage 30 (G, H) or stage 37 (I–L) Xenopus embryos, or 48 hpf zebrafish embryo (T). (M–P) Transverse sections of stage 25 (M, N) or stage 37 (O, P) Xenopus embryos. (Q–T) Dorsal views of four somites (Q) or 24 hpf (R, S) zebrafish embryos. Embryos were injected with 50–100 pg of Xenopus or zebrafish Osr mRNAs. Xenopus embryos were co-injected with 300 pg of LacZ mRNA as a lineage tracer. (A–D) Embryos injected with XOsr1 mRNA showed ectopic patches of Xlim1 (A, B) or XPax8 (C, D) expression in the injected sides (arrowheads in panels B, D). In addition, many embryos have enlarged pronephros (arrows in panels B and D; compare with control sides in panels A and C). (E, F) Stage 25 Xenopus embryos injected with XOsr2 (E) or zOsr1 (F) mRNAs and doubly hybridized for Xlim1 and XPax8. The first chromogenic reaction, to detect Xlim1 expression, is shown in the main panels (cyan), and the second chromogenic reaction, to detect XPax8, in the insets (purple). Note that the same cells express ectopically both markers (arrowheads). (G, H) Embryos injected with 100 pg of Xenopus Osr1 mRNA showed ectopic patches of XSGLT1K (G, arrowheads) and XNKCC2 (H, arrowhead). Note that these embryos have gastrulated properly. (I, J) Enlarged (arrow) and ectopic (arrowhead) kidney tissue, as determined by 3G8 staining, in stage 37 Xenopus embryos injected with XOsr1 (I) or XOsr2 (J) mRNAs. Insets show magnification of ectopic renal tissue in other injected embryos. (K, L) Stage 37 Xenopus embryos co-injected with MOXOsr1 and MTXOsr1 mRNA (K) or MOXOsr2 and MTXOsr2 mRNA (L) and stained for 3G8 monoclonal antibody. Note that these MO insensitive mRNAs rescue the MO-induced kidney marker reduction (arrow) (see panels K and O in Fig. 3 for comparison) and promote ectopic renal tissue (arrowhead). (M–P) Transverse sections on stage 25 (M, N) or stage 37 (O, P) Xenopus embryos injected with XOsr1 (M, O) or XOsr2 (N, P) mRNAs. The embryos in panels M and N show a triple staining for Xlim1 (pronephros, purple), monoclonal antibody 12/101(somitic muscles, brown) and Sox2 (neural tube, magenta). The embryos in panels O, P show differentiated kidneys labeled with the monoclonal antibody 3G8. Note that the ectopic renal tissue is always found close to the neural tube (arrowhead). In addition, these embryos show a clear enlargement of the neural tube and the endogenous pronephros (arrows). (Q–T) Zebrafish embryos injected with zOsr1 (Q, R) or zOsr2 (S, T) mRNAs showing zlim1 expression at 4-somite stage (Q), zPax2.1 expression at 24 hpf (R, S) and differentiated renal structures, as determined by 3G8 monoclonal antibody staining (T). Note the enlarged pronephros (arrowheads) and the ectopic renal tissue (T, arrow). Insets in panels Q, R and T show control non-injected embryos. Image published in: Tena JJ et al. (2007) Copyright © 2007. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View |