02736nas a2200265   4500008004600000245010700046210006900153300001400222490000800236520184900244653001602093653003202109653001202141653002302153653001602176653002402192653001802216653004502234653003602279653003902315100002302354700002102377700002202398856005002420       Submitted                   eng d     00aDesiccation and rehydration elicit distinct heat shock protein transcript responses in flesh fly pupae0 aDesiccation and rehydration elicit distinct heat shock protein t  a963 - 9710 v2073 aHeat shock proteins (Hsps) are a ubiquitous component of the cellular response to stress in both prokaryotic and eukaryotic organisms, but their role and function during desiccation stress in terrestrial arthropods has received limited attention. Molecular responses to rehydration are arguably as important as those to desiccation in maintaining cellular integrity and enzyme activity, but the role of Hsps during stress recovery is poorly understood and has never been addressed with respect to rehydration in insects. This study identifies distinct differences in the Hsp response to desiccation and rehydration in the flesh fly Sarcophaga crassipalpis, as well as differences in the desiccation responses of diapausing and nondiapausing pupae. In nondiapausing pupae, the expression of two inducible Hsps (Hsp23 and Hsp70) is upregulated by desiccation, but the water loss threshold for Hsp expression changes at different rates of dehydration. Continued desiccation results in the prolonged expression of both Hsp23 and Hsp70, which may contribute to the delayed adult eclosion noted in samples desiccated for more than 3 days at &lt;5% relative humidity/25 degrees C. In diapausing pupae, hsp23 and hsp70 transcripts are already highly expressed and are not further upregulated by desiccation stress. Both of the constitutive Hsps investigated, Hsp90 and Hsc70, were unresponsive to desiccation in both nondiapausing and diapausing pupae. However, both Hsp90 and Hsc70 were upregulated upon rehydration in nondiapausing and diapausing pupae. These results indicate distinct roles for the different Hsps during desiccation stress and rehydration/stress recovery. The response to desiccation recovery (rehydration) is similar to the Hsp response to cold recovery identified in S. crassipalpis: Hsp90 and Hsc70 are upregulated in both cases.10a*Body Water10a*Gene Expression Regulation10aAnimals10aBlotting, Northern10aDesiccation10aDiptera/*physiology10aFluid Therapy10aHeat-Shock Proteins/genetics/*metabolism10aHibernation/genetics/physiology10aRNA, Messenger/genetics/metabolism1 aHayward, S., A. L.1 aRinehart, J., P.1 aDenlinger, D., L.  uhttps://sarcophagidae.myspecies.info/node/76402248nas a2200265   4500008004600000245010000046210006900146300001300215490000700228520133300235653003201568653001201600653004701612653003201659653004301691653003801734653005201772653003001824100001501854700001701869700001601886700001401902700001601916856005001932       Submitted                   eng d     00aGene expression responses in larvae of the fleshfly Sarcophaga bullata after immune stimulation0 aGene expression responses in larvae of the fleshfly Sarcophaga b  a98 - 1060 v553 aInsect larvae develop in decaying organic matter and their defence against various microorganisms must therefore be highly efficient. In the present study, we explored the transcriptional kinetics and induction levels of eight genes in Sarcophaga bullata larvae after infection or aseptic injury. Using real-time PCR, we studied the time-dependent immune response of larvae of the fleshfly S. bullata. We compared the mRNA levels of eight selected genes in induced and non-induced larvae. The third-instar larvae of S. bullata were induced by injecting a bacterial suspension of Escherichia coli, Staphylococcus aureus or Pseudomonas aeruginosa, or by simple aseptic injury with an entomological pin. We used intact larvae as a control for basal mRNA expression. Total RNA was isolated from the whole body, fat body and haemocytes. We determined the mRNA levels of genes encoding sapecin, transferrin, prophenoloxidase 1 and 2, storage-binding protein, cathe psin L, sarcocystatin, and 26/29 kDa protease. We found that there was massive up-regulation of genes encoding the fleshfly peptide sapecin, as well as the protein transferrin. We also detected down-regulation of, or no change in, the expression of genes that encode prophenoloxidase 1 and 2, storage-binding protein, cathepsin L, sarcocystatin, and 26/29 kDa protease.10a*Gene Expression Regulation10aAnimals10aDiptera/*genetics/*immunology/microbiology10aEscherichia coli/immunology10aLarva/genetics/immunology/microbiology10aPseudomonas aeruginosa/immunology10aReverse Transcriptase Polymerase Chain Reaction10aStaphylococcus/immunology1 aMasova, A.1 aSindelka, R.1 aKubista, M.1 aKindl, J.1 aJiracek, J.  uhttps://sarcophagidae.myspecies.info/node/292