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FIG. 1. Effects of Xwnt3a on anteroposterior neural pattern in Keller explants. (A, B, and C) Whole-mount in situ hybridization of the
panneural RNP clone, pNPG152; (D, E, and F) Whole-mount in situ hybridization of the cement gland marker, XAG-1. (A and D) Control
albino embryos, stage 19 (A) and stage 25 (D). (B and E) Control Keller explants from stage 10– 10.25 embryos previously injected with
bovine prolactin mRNA (B, 15 of 15 sandwiches were RNP-positive, see arrows; E, 12 of 12 sandwiches were XAG-1-positive, see arrow).
(C and F) Keller explants from stage 10– 10.25 embryos injected with Xwnt3a mRNA (C, 22 of 22 sandwiches were RNP-positive, see
arrows; F, 1 of 12 sandwiches was XAG-1-positive). Keller sandwiches isolated from Xwnt3a-injected embryos exhibited reproducibly
shortened neural ectoderm when compared to Keller sandwiches isolated from prolactin-injected embryos. The Keller explants shown in
(B and C) are from pigmented embryos. (G) RT–PCR analysis of Keller explants using the following set of anteroposterior markers, depicted
with anterior to the top, and increasingly posterior genes below: XAG-1, cement gland; XANF-2, anterior pituitary gland; Otx-A, forebrain;
En-2, midbrain–hindbrain boundary; Krox-20, rhombomeres 3 and 5; Xlhbox-6, posterior spinal cord; NCAM, panneural; muscle actin,
axial mesoderm; Ef1-a, RNA loading control; /RT, plus reverse transcriptase, 0RT, minus reverse transcriptase. |
