Pax6-dependence of Six3, Eya1 and Dach1 expression during lens and nasal placode induction
Section snippets
Results and discussion
The Drosophila eye imaginal disc provides a useful model to study the genetic regulatory networks that operate during organogenesis. The demonstration that Drosophila and mouse Pax6 genes are equally capable of inducing ectopic eyes in fly imaginal discs suggests fundamental conservation of the genetic mechanisms controlling vertebrate and insect eye development (Czerny et al., 1999, Quiring et al., 1994, Zuker, 1994). Moreover, loss-of-function mutations of Pax6 across species present
Embryo collection and genotyping
Embryos were staged by counting somites (Theiler, 1989). Dach1, Eya1 and Sey genotyping was performed as described (Davis et al., 2001, Xu et al., 1999, Xu et al., 1997). Primers specific for Eya2 (Eya2TR (5′-TTAGCTTCAGGTAGCTGCTC-3′) and Eya2T71 (5′-ACACATCTTGTCC-CAGAACGCT-3′)) amplify a 230 bp (base pair) product from the wild type allele, while primers Eya2TR and Neo220f (5′-AAGGGCCAGCT-CATTCCTCCCACT-3′) produce a 230 bp amplicon from the mutant allele.
Immunofluorescence
Immunofluorescence was performed as
Acknowledgements
The Pax6 antibody developed by A. Kawakami was obtained from the Developmental Studies Hybridoma Bank (The University of Iowa). We thank Drs Philip Stashenko and Xin Zhang for critical reading of this manuscript. This work was supported by NIH EY10123 (R.M.) and NIH EY12162 (G.O.).
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