By Alexander Johan Nederbragt
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Embryos were obtained as described previously (van den Biggelaar 1977). Embryo fixation and in situ hybridisation Embryos were fixed and stored and in situ hybridisation was performed as described in chapter 2. Digoxigenin labelled RNA probes were synthesized from PCR products using the DIG RNA labelling kit (Roche). For Pv-otx, the Pv-otxFw2/Pv-otxRv2 PCR fragment was used. For Pv-otp, the 794 bp PCR fragment was used. 52 orthodenticle and orthopedia Results Cloning of Pv-otx and Pv-otp An 84 bp fragment of the homeodomain of a putative otx ortholog was obtained by performing PCR on genomic DNA of Patella vulgata with degenerated primers based on the homeobox of known otx genes.
Apparently, additional genes are involved in lophotrochozoan mesoderm differentiation and this raises questions concerning the overall homology of the mesoderm. N. was supported by the Earth and Life Sciences foundation (ALW), which is subsidized by the Netherlands Organisation for Scientific Research (NWO), the Netherlands. OL was supported by “Programme Génome” from Centre National de la Recherche Scientifique, Université Paris-Sud, and the “Ministère de l’Education Nationale, de la Recherche et de l’Enseignement Supérieur”, France.
Later in Drosophila development, the gene is important for muscle formation (Baylies and Bate 1996; Anant et al. 1998; Cripps and Olson 1998). In other animals, twist genes do not function in early mesoderm formation, but function later in development in mesodermal differentiation, particularly in myogenesis 34 twist (Hopwood et al. 1989; Wolf et al. 1991; Gitelman 1997; Corsi et al. 2000). The data discussed above suggest that a common feature of the role of twist genes in deuterostomes and ecdysozoans is mesodermal differentiation.