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Drosophila melanogaster strain:Canton-S

ABSTRACT: Perception of parasitoid wasps accelerates sexual behavior of Drosophila

PROVIDER: PRJNA642090 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13601374_1.fastq.gz	Fastqsanger.gz
	SRR13601374_2.fastq.gz	Fastqsanger.gz
	SRR13601375_1.fastq.gz	Fastqsanger.gz
	SRR13601375_2.fastq.gz	Fastqsanger.gz
	SRR13601376_1.fastq.gz	Fastqsanger.gz

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Project description:The innate immune response of insects relies on several humoral and cellular mechanisms that require the activation of circulating proteases in the hemolymph to be functional. Here, we analyzed the gelatinase and caseinase activities of Drosophila larval hemolymph under normal and pathogenic conditions (bacterial lipopolysaccharides or endoparasitoid Leptopilina boulardi) using in gel zymography. Gelatinase activity was more intense than caseinase activity and qualitative and quantitative variations were observed between D. melanogaster strains and Drosophila species. Mass spectrometry identified a large number of serine proteases in gel bands equivalent to the major gelatinase and caseinase bands and of these, the most abundant and redundant were Tequila and members of the Jonah and Trypsin protease families. However, hemolymph from Tequila null mutant larvae showed no obvious changes in zymographic bands. Nor did we observe any significant changes in hemolymph gelatinases activity 24 h after injection of bacterial lipopolysaccharides or after oviposition by endoparasitoid wasps. These data confirmed that many serine proteases are present in Drosophila larval hemolymph but those with gelatinase and caseinase activity may not change drastically during the immune response.

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Drosophila melanogaster lamellocytes proteome

Project description:In Drosophila melanogaster larval hemolymph, under normal conditions, plasmatocytes and crystal cells represent respectively ~95% and ~5% of hemocytes, while lamellocytes, the third larval cell type, are absent since they are only induced after parasitoid wasp oviposition, their role being the encapsulation-melanization response to eliminate the wasp egg. However, even after induction lamellocytes number remains low, making difficult biochemical studies. Here using the D. melanogaster hopTum-l mutant that constitutively produces a high number of hemocytes, we set up a method to purify lamellocytes and analyzed their major proteins by 2D gel electrophoresis and their biotinylated plasma membrane surface proteins by 1D SDS-PAGE after affinity purification. Mass spectrometry allowed to identify 430 proteins from the 2D spots and 344 from affinity purified proteins, totalizing 639 unique proteins. Known lamellocyte markers such as PPO3 and the integrin myospheroid are among the major proteins and affinity purification led to the detection of other integrins and a large array of integrins associated proteins involved in cell-cell junction formation and function. Overall newly identified proteins indicated that these cells are highly adapted to the encapsulation process but may have also several different physiological functions. This study provides the basis for new lamellocyte studies in vivo and in vitro, and develop markers to search whether different populations coexist, establish their origins and decipher their respective roles in drosophila physiology and immunity.

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Transcriptomic analysis of a symbiotic poxvirus in parasitoid wasps and fly hosts

Project description:Parasitoid wasps of the species Diachasmimorpha longicaudata are associated with a heritable poxvirus, known as DlEPV, that is stored in the venom gland of adult female wasps and transferred to tephritid fly hosts of the wasps during oviposition. We conducted a RNA-seq differential expression analysis to gain insight on how DlEPV can replicate in both wasps and their fly hosts but only cause pathogenic effects during replication in flies. Our analysis revealed that 91.2% (176 of 193) of DlEPV genes showed significant differential expression during peak virus replication in wasp venom glands compared to parasitized flies. Over 80% of DlEPV replication genes were significantly upregulated in wasps, while 79% of DlEPV putative virulence genes were significantly upregulated in fly hosts. These data therefore support a dichotomy of viral function, where virus replication is promoted in wasp tissue and virulence in host tissue. Such a division of viral activity could represent an important adaptation to maintain a stable symbiosis between this virus and its associated parasitoid.

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