Characterization of the adult Aedes aegypti early midgut peritrophic matrix proteome using LC-MS

Whiten, Shavonn Reezale; Ray, W. Keith; Helm, Richard F.; Adelman, Zach N.

doi:10.1371/journal.pone.0194734

Cited by 26 publications

(38 citation statements)

References 55 publications

(95 reference statements)

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“…Two CTLD-S genes (AAEL000533 and AAEL000556) were observed to be specifically expressed in adult females. Indeed, the corresponding proteins were previously identified as specific to the female salivary glands [ 26 ] and were recovered with other salivary components in the guts of newly fed females [ 32 ]. An additional four CTLD-S genes (AAEL025598, AAEL008681, AAEL004679, and AAEL022823) were all highly expressed, specifically in the testes.…”

Section: Resultsmentioning

confidence: 99%

The C-Type Lectin Domain Gene Family in Aedes aegypti and Their Role in Arbovirus Infection

Adelman

Myles

2018

Viruses

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Several medically important flaviviruses that are transmitted by mosquitoes have been shown to bind to the C-type lectin fold that is present in either vertebrate or invertebrate proteins. While in some cases this interaction is part of a neutralizing anti-viral immune response, many reports have implicated this as critical for successful virus entry. Despite the establishment of mosquito C-type lectin domain containing proteins (CTLDcps) as known host factors in assisting the infectious process for flaviviruses, little is known about the structural characteristics of these proteins and their relationships to each other. In this report, we describe the manual annotation and structural characterization of 52 Aedes aegypti CTLDcps. Using existing RNAseq data, we establish that these genes can be subdivided into two classes: those highly conserved with expression primarily in development (embryo/early larvae) and those with no clear orthologs with expression primarily in late larvae/pupae or adults. The latter group contained all CTLDcps that are regulated by the Toll/Imd immune pathways, all known microbiome-regulating CTLDcps, and almost all CTLDcps that are implicated as flavivirus host factors in A. aegypti. Finally, we attempt to synthesize results from multiple conflicting gene expression profiling experiments in terms of how flavivirus infection changes steady-state levels of mRNA encoding CTLDcps.

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Section: Resultsmentioning

confidence: 99%

The C-Type Lectin Domain Gene Family in Aedes aegypti and Their Role in Arbovirus Infection

Adelman

Myles

2018

Viruses

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“…This hypothesis could be tested through a proteomic analysis at various time points post heme exposure, though such an analysis would be limited to the detection of overall changes in protein abundance and thus could not distinguish all potential protein regulatory changes. A proteomic analysis was recently performed in this mosquito targeting the peritrophic matrix and its heme binding proteins [34], the protocol of which could be adapted here to examine midgut epithelial cells instead. Otherwise, gain of function studies could be performed in a model organism such as yeast [47] or C. elegans that has been modified to temporarily remove native heme import functionality [48].…”

Section: Discussionmentioning

confidence: 99%

“…aegypti midgut epithelial cells are directly involved in heme uptake and digestion following blood feeding [33], we decided to examine their transcriptional responses upon heme exposure. As a typical blood meal taken in by the female mosquito is composed of many different components present in vertebrate blood, we attempted to disentangle their effects on the midgut by using a protein-free artificial bloodmeal [with either 0 mM or 10 mM heme; protocol from [34]]. We initially chose 10 mM heme concentration as that would be the maximum amount of heme present in a drop of blood that could be released at once [18], however smaller concentrations as low as 625 μM were also examined.…”

Section: Znmp Uptake In Aedes Aegypti Midgutsmentioning

confidence: 99%

Transcriptomic analyses of Aedes aegypti cultured cells and ex vivo midguts in response to an excess or deficiency of heme: a quest for transcriptionally-regulated heme transporters

Eggleston

Adelman

2020

BMC Genomics

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Background: Aedes aegypti is the principle vector of many arboviruses, including dengue virus and Zika virus, which are transmitted when an infected female mosquito takes a blood meal in order to initiate vitellogenesis. During blood digestion,~10 mM heme-iron is ingested into the midgut lumen. While heme acts as both a nutrient and signaling molecule during blood digestion, it can also be highly toxic if left unchaperoned. Both signaling by, and degradation of, heme are intracellular processes, occurring in the nucleus and cytoplasm, respectively. However, the precise mechanism of heme uptake into the midgut epithelium is not currently known. Results: We used next generation RNA sequencing with the goal to identify genes that code for membrane bound heme import protein(s) responsible for heme uptake into the midgut epithelium. Heme deprivation increased uptake of a heme fluorescent analog in cultured cells, while treatment of midguts with an excess of heme decreased uptake, confirming physiological changes were occurring in these heme-sensitive cells/tissues prior to sequencing. A list of candidate genes was assembled for each of the experimental sample sets, which included Aag2 and A20 cultured cells as well as midgut tissue, based on the results of a differential expression analysis, soft cluster analysis and number of predicted transmembrane domains. Lastly, the functions related to heme transport were examined through RNAi knockdown. Conclusions: Despite a large number of transmembrane domain containing genes differentially expressed in response to heme, very few were highly differentially expressed in any of the datasets examined. RNAi knockdown of a subset of candidates resulted in subtle changes in heme uptake, but minimal overall disruption to blood digestion/egg production. These results could indicate that heme import in Ae. aegypti may be controlled by a redundant system of multiple distinct transport proteins. Alternatively, heme membrane bound transport in Ae. aegypti could be regulated post-translationally.

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“…6 a-b). As salivary gland proteins can be ingested and potentially impact blood digestion or nutrient absorption [ 38 ], both the fecundity and fertility of SGS1 ko mosquitoes were also evaluated. Both SGS1 ko and LVP females produced the same number of eggs (Fig.…”

Section: Resultsmentioning

confidence: 99%

Aedes aegypti SGS1 is critical for Plasmodium gallinaceum infection of both the mosquito midgut and salivary glands

Kojin

Martín-Martín

Araújo

et al. 2021

Malar J

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Background The invasion of the mosquito salivary glands by Plasmodium sporozoites is a critical step that defines the success of malaria transmission and a detailed understanding of the molecules responsible for salivary gland invasion could be leveraged towards control of vector-borne pathogens. Antibodies directed against the mosquito salivary gland protein SGS1 have been shown to reduce Plasmodium gallinaceum sporozoite invasion of Aedes aegypti salivary glands, but the specific role of this protein in sporozoite invasion and in other stages of the Plasmodium life cycle remains unknown. Methods RNA interference and CRISPR/Cas9 were used to evaluate the role of A. aegypti SGS1 in the P. gallinaceum life cycle. Results Knockdown and knockout of SGS1 disrupted sporozoite invasion of the salivary gland. Interestingly, mosquitoes lacking SGS1 also displayed fewer oocysts. Proteomic analyses confirmed the abolishment of SGS1 in the salivary gland of SGS1 knockout mosquitoes and revealed that the C-terminus of the protein is absent in the salivary gland of control mosquitoes. In silico analyses indicated that SGS1 contains two potential internal cleavage sites and thus might generate three proteins. Conclusion SGS1 facilitates, but is not essential for, invasion of A. aegypti salivary glands by P. gallinaceum and has a dual role as a facilitator of parasite development in the mosquito midgut. SGS1 could, therefore, be part of a strategy to decrease malaria transmission by the mosquito vector, for example in a transgenic mosquito that blocks its interaction with the parasite.

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Characterization of the adult Aedes aegypti early midgut peritrophic matrix proteome using LC-MS

Cited by 26 publications

References 55 publications

The C-Type Lectin Domain Gene Family in Aedes aegypti and Their Role in Arbovirus Infection

The C-Type Lectin Domain Gene Family in Aedes aegypti and Their Role in Arbovirus Infection

Transcriptomic analyses of Aedes aegypti cultured cells and ex vivo midguts in response to an excess or deficiency of heme: a quest for transcriptionally-regulated heme transporters

Aedes aegypti SGS1 is critical for Plasmodium gallinaceum infection of both the mosquito midgut and salivary glands

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