ZOONET: Perspectives on the evolution of animal form. Meeting report

What drives evolution? This was one of the main questions raised at the final ZOONET meeting in Budapest, Hungary, in November 2008. The meeting marked the conclusion of ZOONET, an EU-funded Marie-Curie Research Training Network comprising nine research groups from all over Europe (Max Telford, University College London; Michael Akam, University of Cambridge; Detlev Arendt, EMBL Heidelberg; Maria Ina Arnone, Stazione Zoologica Anton Dohrn Napoli; Michalis Averof, IMBB Heraklion; Graham Budd, Uppsala University; Richard Copley, University of Oxford; Wim Damen, University of Cologne; Ernst Wimmer, University of Göttingen). ZOONET meetings and practical courses held during the past four years provided researchers from diverse backgrounds--bioinformatics, phylogenetics, embryology, palaeontology, and developmental and molecular biology--the opportunity to discuss their work under a common umbrella of evolutionary developmental biology (Evo Devo). The Budapest meeting emphasized in-depth discussions of the key concepts defining Evo Devo, and bringing together ZOONET researchers with external speakers who were invited to present their views on the evolution of animal form. The discussion sessions addressed four main topics: the driving forces of evolution, segmentation, fossils and phylogeny, and the future of Evo Devo

Sexual dimorphism and natural variation within and among species in the Drosophila retinal mosaic

Background Insect compound eyes are composed of ommatidia, which contain photoreceptor cells that are sensitive to different wavelengths of light defined by the specific rhodopsin proteins that they express. The fruit fly Drosophila melanogaster has several different ommatidium types that can be localised to specific retinal regions, such as the dorsal rim area (DRA), or distributed stochastically in a mosaic across the retina, like the ‘pale’ and ‘yellow’ types. Variation in these ommatidia patterns very likely has important implications for the vision of insects and could underlie behavioural and environmental adaptations. However, despite the detailed understanding of ommatidia specification in D. melanogaster, the extent to which the frequency and distribution of the different ommatidium types vary between sexes, strains and species of Drosophila is not known. Results We investigated the frequency and distribution of ommatidium types based on rhodopsin protein expression, and the expression levels of rhodopsin transcripts in the eyes of both sexes of different strains of D. melanogaster, D. simulans and D. mauritiana. We found that while the number of DRA ommatidia was invariant, Rh3 expressing ommatidia were more frequent in the larger eyes of females compared to the males of all species analysed. The frequency and distribution of ommatidium types also differed between strains and species. The D. simulans strain ZOM4 has the highest frequency of Rh3 expressing ommatidia, which is associated with a non-stochastic patch of pale and odd-coupled ommatidia in the dorsal-posterior of their eyes. Conclusions Our results show that there is striking variation in the frequency and distribution of ommatidium types between sexes, strains and species of Drosophila. This suggests that evolutionary changes in the underlying regulatory mechanisms can alter the distribution of ommatidium types to promote or restrict their expression in specific regions of the eye within and between species, and that this could cause differences in vision among these flies

The wnt and delta-notch signalling pathways interact to direct pair-rule gene expression via caudal during segment addition in the spider Parasteatoda tepidariorum

In short germ arthropods, posterior segments are added sequentially from a growth zone or segment addition zone (SAZ) during embryogenesis. Studies in spiders such as the common house spider, Parasteatoda tepidariorum, have provided insights into the gene regulatory network (GRN) that underlies the development of the SAZ, and revealed the involvement of two important signalling pathways. It was shown that Wnt8 maintains a pool of undifferentiated cells in the SAZ, but this ligand is also required for dynamic Delta (Dl) expression associated with the formation of new segments. However, it remains unclear how these pathways interact during SAZ formation and subsequently regulate segment addition. Here we show that Delta-Notch signalling is required for Wnt8 expression in posterior SAZ cells, but represses the expression of this Wnt gene in anterior SAZ cells. We also found that these two signalling pathways are required for the expression of the spider orthologues of the segmentation genes even-skipped (eve) and runt-1 (run-1), at least in part via the transcription factor encoded by caudal (cad). Moreover, it appears that dynamic expression of eve in this spider does not require a feedback loop with run-1, as is found in the pair-rule circuit of the beetle Tribolium. Taken together, our results suggest that the development of posterior segments in Parasteatoda is directed by dynamic interactions between Wnt8 and Delta-Notch signalling that are read out by cad, which is necessary but not sufficient to regulate the expression of the pair-rule genes eve and run-1. Our study therefore provides new insights towards better understanding the evolution and developmental regulation of segmentation in other arthropods including insects

A comprehensive reference transcriptome resource for the common house spider Parasteatoda tepidariorum

Parasteatoda tepidariorum is an increasingly popular model for the study of spider development and the evolution of development more broadly. However, fully understanding the regulation and evolution of P. tepidariorum development in comparison to other animals requires a genomic perspective. Although research on P. tepidariorum has provided major new insights, gene analysis to date has been limited to candidate gene approaches. Furthermore, the few available EST collections are based on embryonic transcripts, which have not been systematically annotated and are unlikely to contain transcripts specific to post-embryonic stages of development. We therefore generated cDNA from pooled embryos representing all described embryonic stages, as well as post-embryonic stages including nymphs, larvae and adults, and using Illumina HiSeq technology obtained a total of 625,076,514 100-bp paired end reads. We combined these data with 24,360 ESTs available in GenBank, and 1,040,006 reads newly generated from 454 pyrosequencing of a mixed-stage embryo cDNA library. The combined sequence data were assembled using a custom de novo assembly strategy designed to optimize assembly product length, number of predicted transcripts, and proportion of raw reads incorporated into the assembly. The de novo assembly generated 446,427 contigs with an N50 of 1,875 bp. These sequences obtained 62,799 unique BLAST hits against the NCBI non-redundant protein data base, including putative orthologs to 8,917 Drosophila melanogaster genes based on best reciprocal BLAST hit identity compared with the D. melanogaster proteome. Finally, we explored the utility of the transcriptome for RNA-Seq studies, and showed that this resource can be used as a mapping scaffold to detect differential gene expression in different cDNA libraries. This resource will therefore provide a platform for future genomic, gene expression and functional approaches using P. tepidariorum

Genetic and developmental analysis of differences in eye and face morphology between Drosophila simulans and Drosophila mauritiana

Eye and head morphology vary considerably among insects and even between closely related species of Drosophila. Species of the D. melanogaster subgroup, and other Drosophila species, exhibit a negative correlation between eye size and face width (FW); for example, D. mauritiana generally has bigger eyes composed of larger ommatidia and conversely a narrower face than its sibling species. To better understand the evolution of eye and head morphology, we investigated the genetic and developmental basis of differences in eye size and FW between male D. mauritiana and D. simulans. QTL mapping of eye size and FW showed that the major loci responsible for the interspecific variation in these traits are localized to different genomic regions. Introgression of the largest effect QTL underlying the difference in eye size resulted in flies with larger eyes but no significant difference in FW. Moreover, introgression of a QTL region on the third chromosome that contributes to the FW difference between these species affected FW, but not eye size. We also observed that this difference in FW is detectable earlier in the development of the eye-antennal disc than the difference in the size of the retinal field. Our results suggest that different loci that act at different developmental stages underlie changes in eye size and FW. Therefore, while there is a negative correlation between these traits in Drosophila, we show genetically that they also have the potential to evolve independently and this may help to explain the evolution of these traits in other insects

Genetics and mechanics combine to guide the embryonic gut

During embryonic development, generating the correct 3D body form, a process called morphogenesis, requires extensive tissue remodelling. Sheets of cells fold and alter their geometry, undergoing changes equivalent to the paper-folding intricacies of origami. In an early embryo, the cells that will form muscle tissue (termed the mesoderm) and gut tissue (the endoderm) move inwards, and the cells of the outer layer form the skin. Writing in Nature, Bailles et al.1 report a previously unknown aspect of how cells internalize, as revealed by studies of the fruit fly Drosophila melanogaster