Dataset Information

Comparative Analysis of Midgut Regeneration Capacity and Resistance to Oral Infection in Three Disease-Vector Mosquitoes.

ABSTRACT: Mosquitoes acquire the pathogens they transmit through ingestion, and the insects' gut constitutes the first line of defense against invading pathogens. Indeed the gut epithelium acts as a physical barrier, activates local antimicrobial peptides production and triggers the systemic immune response. Consequently, gut epithelium is constantly confronted to stress and often suffers cellular damage. We have previously shown that regenerative cells are present in the guts of adult Aedes albopictus, and that chemical damage or bacterial infection leads to the proliferation of these regenerative cells in the midgut. In this study, we extended the analysis of gut cells response to stress to two other important disease vector mosquitoes: Culex pipiens and Anopheles gambiae. We fed mosquitoes on sucrose solutions or on sucrose supplemented with pathogenic bacteria or with damage-inducing chemicals. We also assayed the survival of mosquitoes following the ingestion of pathogenic bacteria. We found that in adult C. pipiens, dividing cells exist in the digestive tract and that these cells proliferate in the midgut after bacterial or chemical damage, similarly to what we previously observed in A. albopictus. In sharp contrast, we did not detect any mitotic cell in the midguts of A. gambiae mosquitoes, neither in normal situation nor after the induction of gut damage. In agreement with this observation, A. gambiae mosquitoes were more sensitive to oral bacterial infections compared to A. albopictus and C. pipiens. This work provides evidence that major differences in gut physiological responses exist between different mosquitoes. The presence of regenerative cells in the mosquito guts and their ability to multiply after gut damage affect the mosquito survival to oral infections, and is also likely to affect its vectorial capacity.

SUBMITTER: Janeh M

PROVIDER: S-EPMC6787257 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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Comparative Analysis of Midgut Regeneration Capacity and Resistance to Oral Infection in Three Disease-Vector Mosquitoes.

Janeh Maria M Osman Dani D Kambris Zakaria Z

Scientific reports 20191010 1

Mosquitoes acquire the pathogens they transmit through ingestion, and the insects' gut constitutes the first line of defense against invading pathogens. Indeed the gut epithelium acts as a physical barrier, activates local antimicrobial peptides production and triggers the systemic immune response. Consequently, gut epithelium is constantly confronted to stress and often suffers cellular damage. We have previously shown that regenerative cells are present in the guts of adult Aedes albopictus, a ...[more]

PMID: 31601867

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Project description:BackgroundSmall RNA regulatory pathways (SRRPs) control key aspects of development and anti-viral defense in metazoans. Members of the Argonaute family of catalytic enzymes degrade target RNAs in each of these pathways. SRRPs include the microRNA, small interfering RNA (siRNA) and PIWI-type gene silencing pathways. Mosquitoes generate viral siRNAs when infected with RNA arboviruses. However, in some mosquitoes, arboviruses survive antiviral RNA interference (RNAi) and are transmitted via mosquito bite to a subsequent host. Increased knowledge of these pathways and functional components should increase understanding of the limitations of anti-viral defense in vector mosquitoes. To do this, we compared the genomic structure of SRRP components across three mosquito species and three major small RNA pathways.ResultsThe Ae. aegypti, An. gambiae and Cx. pipiens genomes encode putative orthologs for all major components of the miRNA, siRNA, and piRNA pathways. Ae. aegypti and Cx. pipiens have undergone expansion of Argonaute and PIWI subfamily genes. Phylogenetic analyses were performed for these protein families. In addition, sequence pattern recognition algorithms MEME, MDScan and Weeder were used to identify upstream regulatory motifs for all SRRP components. Statistical analyses confirmed enrichment of species-specific and pathway-specific cis-elements over the rest of the genome.ConclusionAnalysis of Argonaute and PIWI subfamily genes suggests that the small regulatory RNA pathways of the major arbovirus vectors, Ae. aegypti and Cx. pipiens, are evolving faster than those of the malaria vector An. gambiae and D. melanogaster. Further, protein and genomic features suggest functional differences between subclasses of PIWI proteins and provide a basis for future analyses. Common UCR elements among SRRP components indicate that 1) key components from the miRNA, siRNA, and piRNA pathways contain NF-kappaB-related and Broad complex transcription factor binding sites, 2) purifying selection has occurred to maintain common pathway-specific elements across mosquito species and 3) species-specific differences in upstream elements suggest that there may be differences in regulatory control among mosquito species. Implications for arbovirus vector competence in mosquitoes are discussed.

Project description:Arboviruses are transmitted by specific vectors, and the reasons for this specificity are not fully understood. One contributing factor is the existence of tissue barriers within the vector such as the midgut escape barrier. We used microRNA (miRNA) targeting of Sindbis virus (SINV) to study how replication in midgut cells contributes to overcoming this barrier in the mosquito Aedes aegypti. SINV constructs were designed to be attenuated specifically in midgut cells by inserting binding sites for midgut-specific miRNAs into either the 3' untranslated region (MRE3'miRT) or the structural open reading frame (MRE-ORFmiRT) of the SINV genome. Both miRNA-targeted viruses replicated less efficiently than control viruses in the presence of these miRNAs. When mosquitoes were given infectious blood meals containing miRNA-targeted viruses, only around 20% (MRE3'miRT) or 40% (MRE-ORFmiRT) of mosquitoes developed disseminated infection. In contrast, dissemination occurred in almost all mosquitoes fed control viruses. Deep sequencing of virus populations from individual mosquitoes ruled out selection for mutations in the inserted target sequences as the cause for dissemination in these mosquitoes. In mosquitoes that became infected with miRNA-targeted viruses, titers were equivalent to those of mosquitoes infected with control virus in both the midgut and the carcass, and there was no evidence of a threshold titer necessary for dissemination. Instead, it appeared that if infection was successfully established in the midgut, replication and dissemination were largely normal. Our results support the hypothesis that replication is an important factor in allowing SINV to overcome the midgut escape barrier but hint that other factors are also likely involved. IMPORTANCE When a mosquito ingests an arbovirus during a blood meal, the arbovirus must escape from the midgut of the vector and infect the salivary glands in order to be transmitted to a new host. We used tissue-specific miRNA targeting to examine the requirement for Sindbis virus (SINV) to replicate in midgut epithelium in order to cause disseminated infection in the mosquito Aedes aegypti. Our results indicate that specifically reducing the ability of SINV to replicate in the mosquito midgut reduces its overall ability to establish infection in the mosquito, but if infection is established, replication and dissemination occur normally. These results are consistent with an importance for replication in the midgut epithelium in aiding arboviruses in crossing the midgut barrier.

Dataset Information

Comparative Analysis of Midgut Regeneration Capacity and Resistance to Oral Infection in Three Disease-Vector Mosquitoes.

Publications

Comparative Analysis of Midgut Regeneration Capacity and Resistance to Oral Infection in Three Disease-Vector Mosquitoes.

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