XB-IMG-82120
Xenbase Image ID: 82120
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Fig. 1. ADMP inhibits head formation through repression of head inducers. (A) Embryos were radially injected in the marginal zone at the four-cell stage with morpholino antisense oligo (MO): Control MO (80 ng/embryo), BMP4 and BMP7 MOs (mix of 25 ng each/embryo) or ADMP MO (80 ng/embryo). Embryos were stained by in situ hybridization for
neural markers (mix of xAg1, xRx-1, Krox20, and
HoxB9) at late neurula stage (A), for xRx-1 at early
neurula stage (D; anterior view), or for Sizzled at
gastrula stage (G; ventral view, dorsal up; Nieuwkoop
and Faber stage 10.5). Number of embryos (n)
was ≥18, derived from two independent experiments;
for each class the frequency of the shown
phenotype was >80%. (J) Similar in situ hybridization
to A of embryos injected with ADMP mRNA
(15 pg/embryo) or BMP2 mRNA (15 pg/embryo).
Insets in J show the close-up of head/anterior region
in ventral view. Number of embryos (n) was
≥20, derived from two independent experiments; for
each class the frequency of the shown phenotype
was >80%. (P) Cerberus (Cer) and DKK-1 expression
was monitored by qPCR on dorsal and ventral halves
of embryos injected with ADMP, BMP4/7, or control
morpholino (CoMO). Expression levels are normalized
to EF1a mRNA. (Q) Expression of head-organizer
markers in anterior endodermal cells is inhibited
by ADMP. Four-cell stage embryos were injected with
ADMP MO (80 ng) or control MO (CoMO, 80 ng). At
stage 11, embryos were fixed, removed of the blastocele
roof to reveal the underlying endoderm (for Q
and R), and processed for in situ hybridization for the
indicated markers. Number of embryos (n) was ≥20,
derived from two independent experiments; for each
marker the frequency of the shown expression was
>70%. For additional markers, refer to Fig. S1 G.
(U, Left) Thirty-twoell-stage embryos shown from
animal pole. Embryos were injected with the Smad1-
responsive reporter ID1-lux in dorsal 1blastomeres.
Targeting to anterior endoderm was confirmed at
gastrula by coinjected GFP mRNA (Lower Left picture). (Right) Luciferase was determined on extracts from stage 10.5 embryos coinjected with control or
ADMP MO. Image published in: Inui M et al. (2012) Copyright © 2012. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View |