Xenbase Image ID: 139300

Fig. 5. BENI is involved in gastrulation movements and affects the expression of Xbra both in vitro and in vivo, but does not modulate cell differentiation. (A) The BENI mRNA inhibits elongation of animal cap explants in response to activin. Four-cell embryos were injected with the BENI mRNA (1 ng) or the BENI-MO (4 pmol = 34 ng). Animal caps from uninjected or injected embryos were left untreated or treated with 25 ng/ml activin and cultured for 12 h. Control caps remained round. (B) BENI regulates the expression of Xbra in vitro. Four-cell embryos were injected with the BENI mRNA or the BENI-MO. Animal caps from uninjected or injected embryos were left untreated or treated with 25 ng/ml activin and cultured for 3 h. The animal caps were analyzed by RT-PCR. Mix. 1 (mesoendodermal marker), goosecoid, and Chordin (dorsal mesodermal marker), Xbra (pan-mesodermal marker) were examined. ODC served as a loading control. No signal was observed in the absence of reverse transcriptase [RT(−)]. (D) Neither BENI mRNA nor BENI-MO altered the expression of the marker genes. The animal caps were analyzed by RT-PCR. NCAM (neural marker), Hox B9 (spinal chord marker) type II collagen (notochord marker), muscle actin (m-actin) (muscle marker), and endodermin (endoderm marker) were examined. ODC served as a loading control. No signal was observed in the absence of reverse transcriptase [RT(−)]. (E) BENI mRNA reduced the expression of Xbra in vivo. Spatial expression pattern of Xbra at stage 10.5 detected by whole-mount in situ hybridization. BENI mRNA (300 pg) or the BENI-MO (4 pmol = 34 ng), together with lacZ mRNA (300 pg), was injected at the 4-cell stage into both dorsal blastomeres. β-galactosidase activity was visualized with Red-gal substrate as a tracer. Embryos were viewed from the vegetal side. Image published in: Homma M et al. (2007) Copyright © 2007. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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