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Transcriptomics of phagocytosis and melanization in Aedes aegypti.

ABSTRACT: As is the case in vertebrates, successful mosquito immune responses depend on a complex and carefully orchestrated network of processes in order to defend the host from a foreign invader and to simultaneously maintain a homeostatic environment. Although commonalities exist between immune response mechanisms across phyla, invertebrates (e.g., mosquitoes) lack anticipatory, adaptive immune responses. However, a number of immune response elements, both cellular and humoral, exist to combat infection. Organisms that successfully invade the mosquito body cavity (hemocoel) potentially are subjected to a variety of mosquito defenses including: melanotic encapsulation, the production of antimicrobial peptides (AMPs) or reactive oxygen and nitrogen intermediates, or to engulfment by phagocytic cells. These arrays are used to temporally compare and contrast transcriptome profiles associated with these responses, as manifest in the cells (hemocytes) that circulate in the mosquito body cavity. Hemocytes have documented association with phagocytosis and melanization reactions in mosquitoes, and these responses, to a variety of Gram positive and Gram negative bacteria, have been characterized. Inoculated Escherichia coli elicit a strong phagocytic response, and Micrococcus luteus are rapidly melanized. For transcriptome profiling, inoculated moquitoes are compared to naïve (uninoculated) mosquitoes. Data obtained will begin to clarify the interactive role of hemocyte genes and gene products in orchestrating phagocytic, melanization, and AMP responses against invading bacteria. Keywords: time course

ORGANISM(S): Aedes aegypti

PROVIDER: GSE4279 | GEO | 2007/07/01

SECONDARY ACCESSION(S): PRJNA94947

REPOSITORIES: GEO

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Project description:Background: The mosquito Anopheles gambiae is a major vector of human malaria. Increasing evidence indicates that blood cells (hemocytes) comprise an essential arm of the mosquito innate immune response against both bacteria and malaria parasites. To further characterize the role of hemocytes in mosquito immunity, we undertook the first genome-wide transcriptomic analyses of adult female An. gambiae hemocytes following infection by two species of bacteria and a malaria parasite. Results: We identified 4047 genes expressed in hemocytes, using An. gambiae genome-wide microarrays. While 279 transcripts were significantly enriched in hemocytes relative to whole adult female mosquitoes, 959 transcripts exhibited immune challenge-related regulation. The global transcriptomic responses of hemocytes to challenge with different species of bacteria and/or different stages of malaria parasite infection revealed discrete, minimally overlapping, pathogen-specific signatures of infection-responsive gene expression; 105 of these represented putative immunity-related genes including anti-Plasmodium factors. Of particular interest was the specific co-regulation of various members of the Imd and JNK immune signaling pathways during malaria parasite invasion of the mosquito midgut epithelium. Conclusion: Our genome-wide transcriptomic analysis of adult mosquito hemocytes reveals pathogen-specific signatures of gene regulation and identifies several novel candidate genes for future functional studies. In order to identify hemocyte-specific and immune-responsive transcripts, we first compared transcripts expressed in hemocytes from one day old sugar-fed mosquitoes to transcripts detected in whole mosquitoes of the same age and feeding status. This resulted in identification of the hemocyte-enriched transcriptome. We then compared hemocytes from 1 day old mosquitoes, 1 hour after immune challenge with heat-killed Escherichia coli or Micrococcus luteus, to control female mosquitoes injected with sterile PBS to determine the bacteria challenge responsive transcriptomes. We used heat-killed bacteria in these assays, because our primary interest was in identifying the bacterial responsive transcriptome and to avoid the potentially confounding effects of altered gene expression due to the lethal effects of a systemic infection associated with injection of living bacteria. Lastly, we compared hemocytes from mosquitoes at 24 hours and 19 days after ingestion of a blood meal infected with Plasmodium berghei to mosquitoes of the same age fed a non-infected blood meal to determine the ookinete and sporozoite infection responsive transcriptomes, respectively. This design resulted in a total of five experimental treatments. The following samples are not included in this submission: Hemo E coli vs. hemo unchallenged A Hemo E coli vs. hemo unchallenged B Hemo m luteus vs. hemo unchallenged A Hemo m luteus vs. hemo unchallenged B

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Transcriptomics of phagocytosis and melanization in Aedes aegypti.

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