Temporospatial expression of Dmrt1 in chicken urogenital system (Gallus gallus) using whole mount in situ hybridization.

Doublesex and mab-3-related transcription factor 1 (Dmrt1) is a Z-linked gene that putatively determines the phenotype of gonads in birds. The sex differential expression of Dmrt1 was examined using wholemount in situ hybridization (WISH) in the urogenital systems during embryogenesis. The results revealed that Dmrt1 showed dimorphic expression in chicken gonads, which increased from day 6.5 to day 10.5. The expression of Dmrt1 in male (ZZ) gonads was not twice as much as in female (ZW) gonads, suggesting the existence of other regulatory mechanisms in addition to Z chromosome dosage effect.

Different gene transfer methods at the very early, early, late and whole embryonic stages in chicken.

New technologies in gene transfer combined with experimental embryology make the chicken embryo an excellent model system for gene function studies. The techniques of in ovo electroporation, in vitro culture for ex ovo electroporation and retrovirus-mediated gene transfer have already been fully developed in chicken. Yet to our knowledge, there are no definite descriptions on the features and application scopes of these techniques. The survival rates of different in vitro culture methods were compared and the EGFP expression areas of different gene transfer techniques were explored. It was that the optimal timings of removing embryo for EC culture and Petri dish system was at E1.5 and E2.5, respectively; and optimal timing of injecting retrovirus is at E0. Results indicated that the EC culture, in ovo electroporation, the Petri dish system and retrovirus-mediated method are, respectively, suitable for the very early, early, late and whole embryonic stages in chicken. Comparison of different gene transfer methods and establishment of optimal timings are expected to provide a better choice of the efficient method for a particular experiment.