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. 2019 Apr 1;20(6):e45842. doi: 10.15252/embr.201845842

Figure 3. BMP activity regulates neural AP patterning.

Figure 3

  1. One‐cell stage embryos were injected animally with mRNAs encoding the noggin (5/20 pg) and/or meis3 (0.5 ng) proteins. AC explants were removed from control and injected embryos at blastula stage 9, and explants were cultured to neurula stage 17. Total RNA was isolated from seven control embryos (lane 2), and from eighteen ACs from each group (lanes 3–8). Neural AP markers were examined by sqRT–PCR: forebrain (xanf1, otx2), hindbrain (krox20, hoxb1, hoxb3), spinal cord (hoxb9, cdx1‐4), and panneural (soxd). Ef1α is the control for quantitating RNA levels. ‐RT–PCR was performed on total RNA isolated from control embryos (lane 1).
  2. One‐cell stage embryos were injected animally with mRNA encoding bmp4 (0.2 ng) protein. Total RNA was isolated from five control embryos at neurula stage 17 (lane 1) and five embryos from the injected group (lanes 2). Neural and mesodermal markers were examined by sqRT–PCR: forebrain (otx2), hindbrain (krox20, hoxb3), spinal cord (hoxb9, hoxc10, cdx4), panneural (nrp1, ncam), and mesodermal (muscle actin—MA). ODC is the control for quantitating RNA levels. ‐RT–PCR was performed on total RNA isolated from control embryos (lane 1).
  3. One‐cell stage embryos were injected animally with mRNA encoding bmp4 (0.2 ng) protein Embryos underwent in situ hybridization. Expression of the hindbrain marker hoxb3 was reduced/eliminated by bmp4 (0.2 ng) in 68% of the embryos (n = 19). Expression of the spinal cord marker cdx4 was slightly increased/normal, in 100% of the embryos (n = 19).