ScRNA-seq of female Anopheles gambiae mosquito midguts at multiple timepoints after a Plasmodium falciparum-infectious blood meal
Ontology highlight
ABSTRACT:
ORGANISM(S): Anopheles gambiae Plasmodium falciparum
PROVIDER: GSE284537 | GEO | 2025/08/18
REPOSITORIES: GEO
Similar Datasets
Project description:A deeper understanding of malaria parasite development inside the Anopheles mosquito may lead to the identification of processes that can be targeted by transmission-blocking interventions. Paraquat (1,1'-dimethyl-4,4'-bipyridylium dichloride) is a potent superoxide-inducing agent that impacts Plasmodium ookinete development, especially at higher concentrations. Compounds like Paraquat can potentially induce an oxidative imbalance in the mosquito midgut during ookinete maturation, essentially super-stressing the parasite leading to the arrested development of ookinetes, the only stage that can invade through a mosquito midgut cell to establish an oocyst infection in the mosquito. The mosquito midgut has evolved to handle the natural production of reactive oxygen and nitrogen species (ROS and RNS, respectively) as a result of feeding on blood. The addition of Paraquat to a bloodmeal is expected to induce a cognate response in the midgut to handle the excess ROS/RNS, and high concentrations of this compound can potentially overwhelm the midgut response leading to mosquito death. While several studies have explored the effect of Paraquat on malaria parasites, a fundamental understanding of the mosquito response to this compound remains unknown. Here, we quantified the mosquito midgut proteomic response to a Paraquat-laced sugar meal to understand the intrinsic midgut response (in the absence of a bloodmeal). We then carried out transcriptomic analysis of the mosquito midgut for several antioxidants of the Trx and GSH pathways to compare concordance or discordance between protein and its transcripts during different oxidative stress conditions. Finally, we determined whether the same Trx and GSH pathways are upregulated following infection with either P. falciparum or P. berghei at 24 hrs post-blood feeding, coinciding with the time point for maximal ookinete traversal of the midgut. We discuss the potential selective action of Paraquat on the parasite and the intrinsic tolerance of the mosquito midgut to Paraquat-mediated oxidative stress.
2022-03-01 | PXD008231 | Pride
Project description:P. falciparum undergoes antigenic variation during its life cycle in the human host. To accomplish this, the malaria parasite has developed mechanisms that ensure the expression of a single member of the var gene family to produce one of the PfEMP1 variant proteins. Here we carry out RNA-seq and ChIP-seq analyses of histone modifications to explore transcriptional and epigenomic changes taking place between the transmissible stages of the parasite i.e. gametocytes present in human blood and sporozoites present in the mosquito salivary glands. We find that most var genes are expressed at low levels in gametocytes but a single var gene is selected during the oocyst stage in the mosquito. Clonal expression of a specific var gene is accompanied by the establishment of specific histone modification patterns in the active and inactive genes. These modifications are epigenetically transmitted from the oocyst to the infective sporozoite stage, where expression of the active var gene is amplified by the transcription of an antisense lncRNA. The findings suggest a critical role for the mosquito immune system in determining the choice of var gene activation prior to transmission to the human host.
2017-01-18 | GSE68667 | GEO
Project description:Transmission of malaria is dependent on the successful completion of the Plasmodium lifecycle in the Anopheles vector. Major obstacles are encountered in the midgut tissue, where most parasites are killed by the mosquito’s immune system. In the present study, DNA microarray analyses have been used to compare Anopheles gambiae responses to invasion of the midgut epithelium by the ookinete stage of the human pathogen Plasmodium falciparum and the rodent experimental model pathogen P. berghei. Invasion by P. berghei had a more profound impact on the mosquito transcriptome, including a variety of functional gene classes, while P. falciparum elicited a broader immune response at the gene transcript level. Ingestion of human malaria-infected blood lacking invasive ookinetes also induced a variety of immune genes, including several anti-Plasmodium factors. By comparing gene expression patterns between carcassess or guts of mosquitoes that fed on a P. falciparum or P. berghei wt and mosquitoes that fed on invasion incapable strains we gain information on the A. gambiae transcriptional responses to the invading ookinete at 24 hours after feeding. By comparing gene expression patterns between carcassess or guts of mosquitoes that fed on a P. falciparum ookinete invasion incapable strain and mosquitoes that fed on non-infected blood we gain information on the A. gambiae transcriptional responses to malaria infected blood in absence of ookinete invasion at 24 hours after feeding. By comparing gene expression patterns between mosquitoes at 4 hours after being injected with either E. coli or S. aureus and mosquitoes injected with sterile PBS we gain information on the mosquito's transcriptional response to these bacterial challenges.
2007-08-15 | E-GEOD-8789 | biostudies-arrayexpress
Project description:A general understanding of redox homeostasis in An. gambiae midgut epithelial cells under different oxidative conditions during P. falciparum ookinete invasion is missing. We used an ex vivo midgut culture model to understand AgTrx-1 protein expression pattern in An. gambiae midguts under oxidative stress of the ROS inducer, tert-butyl hydroperoxide (tBHP). We then quantified the midgut proteomic expression profile to understand organ-level regulation of the response to oxidative stress. An. gambiae midguts under low (50M) and high (200M) tBHP concentrations were enriched in ribosomal proteins (RPs), consistent with cells undergoing ribosomal/nucleolar stress. Nevertheless, these midguts were also enriched in peptidases, integral membrane proteins, and cytochrome P450s indicating cells that are undergoing protein processing and folding through post-translational modification (PTMs), and detoxification of toxic compounds through active efflux. This response to oxidative stress was strikingly dissimilar to that observed from previous studies with another dipteran, Drosophila following oxidative stress induction. Our data indicates that the response to tBHP induced oxidative stress is mild and suggests that the associated antioxidant response is similar to what is observed during midgut invasion of P. falciparum okinete. Furthermore, our data shows that ribosomal/nucleolar stress is crucial in the regulation of oxidative stress in An. gambiae midguts. Given this importance, its possible to develop mechanisms to perturb this ribosomal/nucleolar stress response in An. gambiae resulting into unmanageable levels of oxidative stress exposure to Plasmodium parasite in the midgut, yet still allowing the mosquito to survive the perturbation. Unmanageable levels of oxidative stress could result to harmful effect to the Plasmodium and eventually its death resulting into halting of its transmission.
2018-11-21 | PXD008251 | Pride
Project description:cGMP signal-activated ookinete gliding is essential for mosquito midgut infection of Plasmodium in malaria transmission. During ookinete development, cGMP synthesizer GCβ polarizes to a unique localization “ookinete extrados site” (OES) until ookinete maturation and activates cGMP signaling for initiating parasite motility. However, the mechanism underlying GCβ translocation from cytosol to OES remains elusive. Here, we used protein proximity labeling to search the GCβ-interacting proteins in ookinetes of the rodent malaria parasite P. yoelii, and found the top hit Sir2A, a NAD+-dependent sirtuin family deacetylase. Sir2A interacts with GCβ throughout ookinete development. In mature ookinetes, Sir2A co-localizes with GCβ at OES in a mutually dependent manner. Parasites lacking Sir2A lose GCβ localization at OES, ookinete gliding, and mosquito infection, phenocopying GCβ deficiency. GCβ is acetylated at gametocytes but is deacetylated by Sir2A for OES localization at mature ookinetes. We further demonstrated that the level of NAD+, an essential co-substrate for sirtuin, increases during the ookinete development. The NAD+ at its maximal level until ookinete maturation promotes Sir2A-catalyzed GCβ deacetylation, ensuring GCβ localization at OES. This study highlights the spatiotemporal coordination of cytosolic NAD+ level and NAD+-dependent Sir2A in regulating GCβ deacetylation and dynamic localization for Plasmodium ookinete gliding.
2024-09-26 | PXD056305 |
Project description:Catalase is a potent antioxidant, likely involved in post-blood meal homeostasis in mosquitoes. This enzyme breaks down H2O2, preventing the formation of the hydroxyl radical (HO*). Quiescins are newly classified sulfhydryl oxidases that bear a thioredoxin motif at the N-terminal and an ERV1-like portion at the C-terminal. These proteins have a major role in generating disulfides in intra- or extracellular environments, and thus participate in redox reactions. In the search for molecules to serve as targets for novel anti-mosquito strategies, we have silenced a catalase and a putative quiescin/sulfhydryl oxidase (QSOX), from the African malaria vector Anopheles gambiae, through RNA interference (RNAi) experiments. We observed that the survival of catalase- and QSOX-silenced insects was reduced over controls following blood digestion, most likely due to the compromised ability of mosquitoes to scavenge and/or prevent damage caused by blood meal-derived oxidative stress. The higher mortality effect was more accentuated in catalase-silenced mosquitoes, where catalase activity was reduced to low levels. Lipid peroxidation was higher in QSOX-silenced mosquitoes suggesting the involvement of this protein in redox homeostasis following a blood meal. This study points to the potential of molecules involved in antioxidant response and redox metabolism to serve as targets of novel anti-mosquito strategies and offers a screening methodology for finding targetable mosquito molecules.
| S-EPMC2673501 | biostudies-literature
Project description:Malaria inflicts the highest rate of morbidity and mortality among the vector-borne diseases. The dramatic bottleneck of parasite numbers that occurs in the gut of the obligatory mosquito vector provides a promising target for novel control strategies. Using single-cell transcriptomics, we analyzed Plasmodium falciparum development in the mosquito gut, from unfertilized female gametes through the first 20 hours post blood feeding, including the zygote and ookinete stages. This study revealed the temporal gene expression of the ApiAP2 family of transcription factors, and of parasite stress genes in response to the harsh environment of the mosquito midgut. Further, employing structural protein prediction analyses we found several upregulated genes predicted to encode intrinsically disordered proteins (IDPs), a category of proteins known for their importance in regulation of transcription, translation and protein-protein interactions. IDPs are known for their antigenic properties and may serve as suitable targets for antibody or peptide-based transmission suppression strategies. In total, this study uncovers the P. falciparum transcriptome from early-to-late parasite development in the mosquito midgut, inside its natural vector, which provides an important resource for future malaria transmission-blocking initiatives. Single-cell data can be visualized interactively via https://mubasher-mohammed.shinyapps.io/shinyapp/ In-house bash, R code scripts and data that were implemented in this study are available on GitHub https://github.com/ANKARKLEVLAB/Single-cell-P.falciparum-midgut .
2023-02-16 | GSE222586 | GEO
Project description:There were three primary objectives to the overall experiment. I. Describe the transcriptome of the oocyst through its development beginning with the freshly excreted, unsporulated oocyst at “Day 0” to a fully sporulated and mature oocyst at “Day 10” and including a mid-sporulation timepoint at “Day 4”. II. Compare the transcriptomes of in vitro vs. in vivo derived bradyzoites. III. Compare expression data from three life stages (oocyst, bradyzoite and tachyzoite) from the same parasite isolate, M4, which has been been characterized as a Type II strain.
2011-10-01 | GSE32427 | GEO
Project description:The Anopheles gambiae midgut harbors bacteria that proliferate upon a blood feed. We used microarrays to examine the midgut gene expression response at early stages (3hours) after an artifitial meal containing heat killed bacteria.
2010-02-13 | GSE20204 | GEO
Project description:Malaria parasites have a complex life cycle comprising development in two hosts, the vertebrate and the vector mosquito. In the gut of the mosquito, the parasite develops into the oocyst, which is settled beneath the epithelium and attached to the basal lamina of the gut until the maturation of the cyst and its rupture concomitant with the release of the sporozoites, the infectious form of the parasite. The oocyst represents the longest stage of the parasite life cycle but it is poorly understood, mainly because of the difficulties to separate the oocysts from the mosquito midgut tissue but also the lack of a robust method to reproduce this stage in vitro. Here we describe a simple and reproducible protocol for purification of oocysts from mosquitoes. Midguts were dissected from infected mosquitoes and treated with trypsin which resulted in the degradation of the basal lamina and the release of the oocysts from the midgut tissue. The results obtained showed that the isolated oocysts were free of the mosquito protein E-cadherin. Purified oocysts were alive as judged by a strong GFP signal at least up to 2 h after treatment and furthermore sporozoites that had developed in the cyst were able to glide. Our new method will allow the study of the oocyst composition, formation and development in more details leading to advances in knowledge of this Plasmodium stage.
| S-EPMC7190746 | biostudies-literature