One generalist or several specialist species? Wide host range and diverse manipulations of the hosts' web-building behaviour in the true spider parasitoid Zatypota kauros (Hymenoptera: Ichneumonidae)

Parasitoid wasps of the Polysphincta genus-group are highly specialised on their spider hosts, and most of them are known to manipulate their hosts into building a special web in which the parasitoid pupates. Trophic niche and the plasticity of host use were investigated in the koinobiont parasitoid Zatypota kauros Gauld from Queensland, Australia. We found that Z. kauros attacks spider hosts from different families, each differing widely in their web-building behaviours, which makes it unique in the breadth of its host range. Molecular analyses revealed that the taxon Z. kauros contains three divergent mitochondrial lineages. Lineage A was associated exclusively with spiders of the genus Anelosimus (Theridiidae), which builds tangle webs; lineage B was associated with the genus Cyrtophora (Araneidae), which weaves tent webs; and lineage C was associated with a broad range of hosts, including spiders of both the families Araneidae and Theridiidae. Unique host manipulations could be observed in the web-building behaviours of the different host groups. Nevertheless, nuclear data from two ribosomal genes and three introns did not add any support to the existence of different evolutionary lineages, nor did they coincide with the different host groups. The partial correspondence of mitochondrial lineage and host use, together with an apparent mito-nuclear conflict might indicate maternal effects or very recent and/or incomplete speciation in this taxon. Given their wide host range and intriguing interactions with their hosts, the Z. kauros complex represents a promising system for studying parasitoid specialisation and its potential impact on speciation

Identification of AGR2 Gene-Specific Expression Patterns Associated with Epithelial-Mesenchymal Transition

The TGF-β signaling pathway is involved in numerous cellular processes, and its deregulation may result in cancer development. One of the key processes in tumor progression and metastasis is epithelial to mesenchymal transition (EMT), in which TGF-β signaling plays important roles. Recently, AGR2 was identified as a crucial component of the cellular machinery responsible for maintaining the epithelial phenotype, thereby interfering with the induction of mesenchymal phenotype cells by TGF-β effects in cancer. Here, we performed transcriptomic profiling of A549 lung cancer cells with CRISPR-Cas9 mediated AGR2 knockout with and without TGF-β treatment. We identified significant changes in transcripts associated with focal adhesion and eicosanoid production, in particular arachidonic acid metabolism. Changes in transcripts associated with the focal adhesion pathway were validated by RT-qPCR of COL4A1, COL4A2, FLNA, VAV3, VEGFA, and VINC mRNAs. In addition, immunofluorescence showed the formation of stress fibers and vinculin foci in cells without AGR2 and in response to TGF-β treatment, with synergistic effects observed. These findings imply that both AGR2 downregulation and TGF-β have a role in focal adhesion formation and cancer cell migration and invasion. Transcripts associated with arachidonic acid metabolism were downregulated after both AGR2 knockout and TGF-β treatment and were validated by RT-qPCR of GPX2, PTGS2, and PLA2G4A. Since PGE2 is a product of arachidonic acid metabolism, its lowered concentration in media from AGR2-knockout cells was confirmed by ELISA. Together, our results demonstrate that AGR2 downregulation and TGF-β have an essential role in focal adhesion formation; moreover, we have identified AGR2 as an important component of the arachidonic acid metabolic pathway