A cell atlas of the adult Aedes aegypti midgut revealed by single nucleus RNA-Seq
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ABSTRACT:
ORGANISM(S): Aedes aegypti
PROVIDER: GSE245622 | GEO | 2023/10/17
REPOSITORIES: GEO
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Project description:Aedes aegypti mosquito, vector for several viral diseases, undergoes significant physiological changes after a blood meal. Through single-cell RNA sequencing and metabolomics, we unveiled dynamic cellular composition and metabolic adaptations within abdominal midgut and fat body tissues. We revealed high cell diversity, specialized in digestion, metabolism, immunity, and reproduction. While the midgut primarily comprises enterocytes, enteroendocrine, cardia and intestinal stem cells, the fat body consists of not only trophocytes and oenocytes, but also a substantial population of hemocytes and fat body-yolk cells (FYC). The fat body, exhibiting a more complex metabolomic profile than the midgut, played a central role in immune and metabolic gene expression, particularly within trophocytes and FYCs. Additionally, insect-specific viruses were detected at the single-cell level, mainly in the midgut at later stages post-blood meal. These findings offer new vector control strategies by targeting specific abdominal cell populations and metabolic pathways involved after a blood meal.
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Project description:This study seeks to understand the mechanism of midgut escape and identify candidate genes and potential biochemical pathways involved in the midgut infection of chikungunya virus (CHIKV) in vector mosquito Aedes aegypti. We conducted a comparative transcriptomic analysis of midgut samples of female mosquitoes,after feeding a saline meal (SM) or a protein meal (PM) containing CHIKV. Our results allow the conclusion that midgut-expressed genes not involved in blood or protein digestion can be identified by substituting either a bloodmeal or PM for a SM. In presence of orally acquired CHIKV in midguts of SM fed mosquitoes, the majority of the upregulated DE genes belonged to the categories immune system and catalytic activity. These genes included several serine-type endopeptidases, trypsins, collagenases, and M1 zinc metalloproteases, which potentially could be involved in the midgut escape mechanism of CHIKV. One of the serine metalloproteinase genes, AeLT, was further analyzed showing strong (MMP) collagenase activity in vitro. Our results present a set of candidate genes potentially responsible for overcoming the arbovirus midgut escape barrier (MEB) in Ae. aegypti.
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Project description:The female mosquito Aedes aegypti requires amino acids and other nutrients like heme and iron from a blood meal to initiate vitellogenesis. Heme is a pro-oxidant molecule that acts as a nutrient, signaling molecule and in large quantities, as a toxin. Ae. aegypti has developed a few strategies to handle heme toxicity, as during a typical meal ~10mM is released into the midgut lumen. These strategies include heme aggregation to the peritrophic matrix and the degradation of heme by heme oxygenase in the cytosol of the midgut epithelium. However, despite the importance of heme as a nutrient and toxin, the mechanism of entry into the midgut epithelial cells is not currently known. As no heme transport proteins in have been identified in any dipteran, heme fluorescent analog studies were performed to visualize changes in expression caused by heme followed by global expression analyses performed in midgut tissues using NGS-based RNA sequencing with the end goal to identify the gene(s) that encode the membrane bound heme import proteins responsible for heme uptake during blood digestion. Examination of differential expression of mRNA transcripts at the gene level, found 65 significant DE genes at the adjusted p-value cut off of 0.0001, 38 of which are TM containing and only 2 of which showed high expression changes, AAEL019570 (-2.04 log2, ~0.243), unknown function, and AAEL000717 (3.91 log2, ~15.03), a protocadherin. This list was further reduced to 16 genes with potential heme import function and 7 genes with potential heme export function by examination of differential expression, number of TM domains and function relating to transport. As very few highly differentially expressed genes were found in the analysis, heme import may be controlled by a redundant system of multiple transport proteins instead of a single highly expressed one. Alternatively, heme transport in Ae. aegypti could be regulated post-translationally.
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Project description:Oral susceptibility of Aedes aegypti mosquitoes to dengue viruses varies between different Aedes species and strains. However, the midgut-specific transcriptional profile that may produce this variation is presently obscure and was the subject of our investigation. The variation in active expression between dengue-2 susceptible (SUS) and refractory (REF) mosquitoes was investigated during the first critical 96 hours after infection Transcriptional profiles were mined from respective guts using the serial analysis of gene expression technique (SAGE) and libraries constructed from midguts obtained from mosquitoes that received a dengue-2 infected blood meal (DENV-2), a non infected blood meal (naive) or a 5% sucrose meal (SM). Here we report that variation between DENV-2 infected libraries versus respective naïve libraries revealed very few transcripts that were common and statistically significant in DENV-2 infected libraries. In addition, the expression profiles among libraries displayed up regulation of antisense transcripts especially in the SUS strain. A strong proclivity towards strain-specificity in differential expression was observed, which suggested an exclusive transcription that is likely up-regulated after DENV-2 infection
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