scholarly journals Deep Sequencing revealed "Plant Like Transcripts" in mosquito Anopheles culicifacies: an Evolutionary Puzzle

2014 ◽  
Author(s):  
Punita Sharma ◽  
Swati Sharma ◽  
Ashwani Mishra ◽  
Tina Thomas ◽  
Tanwee Das De ◽  
...  
Keyword(s):  
Salivary Gland ◽  
Adult Female ◽  
Blood Meal ◽  
Underlying Mechanism ◽  
Pathogen Transmission ◽  
Anopheles Culicifacies ◽  
Plant Host ◽  
Vector Borne Diseases ◽  
Experimental Validations ◽  
Molecular Relationships

As adult female mosquito's salivary gland facilitate blood meal uptake and pathogen transmission e.g. Plasmodium, virus etc., a plethora of research has been focused to understand the mosquito-vertebrate-pathogen interactions. Despite the fact that mosquito spends longer time over nectar sugar source, the fundamental question "how adult female salivary gland manages molecular and functional relationship during sugar vs. blood meal uptake" remains unanswered. Currently, we are trying to understand these molecular relationships under dual feeding conditions in the salivary glands of the mosquito Anopheles culicifacies. During functional annotation of salivary transcriptome database, unexpectedly we discovered a cluster of salivary transcripts encoding plant like proteins. Our multiple experimental validations confirmed that Plant like transcripts (PLTs) are of mosquito origin and may encode functional proteins. A comprehensive molecular analysis of the PLTs and ongoing metagenomic analysis of salivary microbiome provide first evidence that how mosquito may have been benefited from its association with plant host and microbes. Future understanding of the underlying mechanism of the feeding associated molecular responses may provide new opportunity to control vector borne diseases.

scholarly journals A synergistic transcriptional regulation of olfactory genes derives complex behavioral responses in the mosquito Anopheles culicifacies

2017 ◽  
Author(s):  
Tanwee Das De ◽  
Tina Thomas ◽  
Sonia Verma ◽  
Deepak Singla ◽  
Charu Rawal ◽  
...  
Keyword(s):  
Adult Female ◽  
Blood Meal ◽  
Olfactory System ◽  
Transcriptional Profiling ◽  
Behavioral Responses ◽  
Blood Feeding ◽  
Female Mosquito ◽  
Anopheles Culicifacies ◽  
Host Seeking ◽  
Adult Female Mosquito

AbstractDecoding the molecular basis of host seeking and blood feeding behavioral evolution/adaptation in the adult female mosquito may provide an opportunity to design new molecular strategy to disrupt human-mosquito interactions. However, despite the great progress in the field of mosquito olfaction and chemo-detection, little is known that how the sex-specific specialization of the olfactory system enables adult female mosquitoes to derive and manage complex blood feeding associated behavioral responses. A comprehensive RNAseq analysis of prior and post blood meal olfactory system of An. culicifacies mosquito revealed that a minor but unique change in the nature and regulation of key olfactory genes play a pivotal role in managing diverse behavioral responses. Age dependent transcriptional profiling demonstrated that adult female mosquito’s chemosensory system gradually learned and matured to drive the host-seeking and blood feeding behavior at the age of 5-6 days. A zeitgeber time scale expression analysis of Odorant Binding Proteins (OBPs) unravels unique association with a late evening to midnight peak biting time. Blood meal-induced switching of unique sets of OBP genes and Odorant Receptors (ORs) expression coincides with the change in the innate physiological status of the mosquitoes. Blood meal follows up experiments provide enough evidence that how a synergistic and concurrent action of OBPs-ORs may drive ‘prior and post blood meal’ complex behavioral events. Finally, tissue-specific gene expression analysis and molecular modelling predicted two uncharacterized novel sensory appendages proteins (SAP-1 & SAP2) unique to An. culicifacies mosquito and may play a central role in the host-seeking behavior.SignificanceEvolution and adaptation of blood feeding behavior not only favored the reproductive success of adult female mosquito but also make them an important disease vectors. Immediately after emergence, an environmental exposure may favor the broadly tuned olfactory system of mosquitoes to derive complex behavioral responses. But, how these olfactory derived genetic factors manage female specific ‘pre and post’ blood meal associated complex behavioral responses are not well known. We unraveled synergistic actions of olfactory factors governs an innate to prime learning strategy to facilitate rapid blood meal acquisition and downstream behavioral activities. A species-specific transcriptional profiling and an in-silico analysis predict novel ‘sensory appendages protein’, as a unique target to design disorientation strategy against the mosquito Anopheles culicifacies.

scholarly journals Deep sequencing revealed molecular signature of horizontal gene transfer of plant like transcripts in the mosquito Anopheles culicifacies: an evolutionary puzzle

F1000Research ◽  
2015 ◽  
Vol 4 ◽  
pp. 1523 ◽  
Author(s):  
Punita Sharma ◽  
Tanwee Das De ◽  
Swati Sharma ◽  
Ashwani Kumar Mishra ◽  
Tina Thomas ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Horizontal Gene Transfer ◽  
Functional Role ◽  
Underlying Mechanism ◽  
Molecular Signature ◽  
Anopheles Culicifacies ◽  
Genomic Tools ◽  
Plant Genes ◽  
Experimental Validations

In prokaryotes, horizontal gene transfer (HGT) has been regarded as an important evolutionary drive to acquire and retain beneficial genes for their survival in diverse ecologies. However, in eukaryotes, the functional role of HGTs remains questionable, although current genomic tools are providing increased evidence of acquisition of novel traits within non-mating metazoan species. Here, we provide another transcriptomic evidence for the acquisition of massive plant genes in the mosquito,Anopheles culicifacies. Our multiple experimental validations including genomic PCR, RT-PCR, real-time PCR, immuno-blotting and immuno-florescence microscopy, confirmed that plant like transcripts (PLTs) are of mosquito origin and may encode functional proteins. A comprehensive molecular analysis of the PLTs and ongoing metagenomic analysis of salivary microbiome provide initial clues that mosquitoes may have survival benefits through the acquisition of nuclear as well as chloroplast encoded plant genes. Our findings of PLTs further support the similar questionable observation of HGTs in other higher organisms, which is still a controversial and debatable issue in the community of evolutionists. We believe future understanding of the underlying mechanism of the feeding associated molecular responses may shed new insights in the functional role of PLTs in the mosquito.

scholarly journals MicroRNA-8 targets the Wingless signaling pathway in the female mosquito fat body to regulate reproductive processes

2015 ◽  
Vol 112 (5) ◽  
pp. 1440-1445 ◽  
Author(s):  
Keira J. Lucas ◽  
Sourav Roy ◽  
Jisu Ha ◽  
Amanda L. Gervaise ◽  
Vladimir A. Kokoza ◽  
...  
Keyword(s):  
Blood Meal ◽  
Molecular Mechanisms ◽  
Fat Body ◽  
Control Strategies ◽  
Functional Characterization ◽  
Target Prediction ◽  
Underlying Mechanism ◽  
Female Mosquito ◽  
Reproductive Events

Female mosquitoes require a blood meal for reproduction, and this blood meal provides the underlying mechanism for the spread of many important vector-borne diseases in humans. A deeper understanding of the molecular mechanisms linked to mosquito blood meal processes and reproductive events is of particular importance for devising innovative vector control strategies. We found that the conserved microRNA miR-8 is an essential regulator of mosquito reproductive events. Two strategies to inhibit miR-8 function in vivo were used for functional characterization: systemic antagomir depletion and spatiotemporal inhibition using the miRNA sponge transgenic method in combination with the yeast transcriptional activator gal4 protein/upstream activating sequence system. Depletion of miR-8 in the female mosquito results in defects related to egg development and deposition. We used a multialgorithm approach for miRNA target prediction in mosquito 3′ UTRs and experimentally verified secreted wingless-interacting molecule (swim) as an authentic target of miR-8. Our findings demonstrate that miR-8 controls the activity of the long-range Wingless (Wg) signaling by regulating Swim expression in the female fat body. We discovered that the miR-8/Wg axis is critical for the proper secretion of lipophorin and vitellogenin by the fat body and subsequent accumulation of these yolk protein precursors by developing oocytes.

scholarly journals Salivary AsHPX12 influence pre-blood meal associated behavioral properties in the mosquito Anopheles stephensi

2020 ◽  
Author(s):  
Seena Kumari ◽  
Tanwee Das De ◽  
Charu Chauhan ◽  
Jyoti Rani ◽  
Sanjay Tevatiya ◽  
...  
Keyword(s):  
Salivary Gland ◽  
Blood Meal ◽  
Anopheles Stephensi ◽  
Blood Feeding ◽  
Vital Role ◽  
Host Attraction ◽  
Altered Expression ◽  
Rnaseq Data ◽  
Transcriptional Modulation ◽  
Physiological Homeostasis

AbstractIn the adult female mosquito, successful blood meal acquisition is accomplished by salivary glands, which releases a cocktail of proteins to counteract vertebrate host’s immune-homeostasis. However, the biological relevance of many salivary proteins remains unknown. Here, we characterize a salivary specific Heme peroxidase family member HPX12, originally identified from Plasmodium vivax infected salivary RNAseq data of the mosquito Anopheles stephensi. We demonstrate that dsRNA silencing mediated mRNA depletion of salivary AsHPX12 (80-90%), causes enhanced host attraction but reduced blood-meal acquisition abilities, by increasing probing propensity (31%), as well as probing time (100–200s, P<0.0001) as compared to control (35-90s) mosquitoes group. Altered expression of the salivary secretory and antennal proteins may account for an unusual fast release of salivary cocktail proteins, but the slowing acquisition of blood meal, possibly due to salivary homeostasis disruption of AsHPX12 silenced mosquitoes. A parallel transcriptional modulation in response to blood feeding and P. vivax infection, further establish a possible functional correlation of AsHPX12 role in salivary immune-physiology and Plasmodium sporozoites survival/transmission. We propose that salivary HPX12 may have a vital role in the management of ‘pre- and post’-blood meal associated physiological-homeostasis and parasite transmission.Graphical abstractFigure 1:Schematic representation of mosquito’s blood meal acquisition and upshot on blood-feeding after silencing of salivary gland HPX-12. (A) After landing over host skin, mosquito mouthparts (proboscis) actively engaged to search, probe, and pierce the skin followed by a rapid release of the pre-synthesized salivary cocktail, which counteracts the host homeostasis, inflammation, and immune responses, during blood meal uptake. (B) Silencing of HPX-12 disrupts salivary gland homeostasis, enhancing mosquito attraction, possibly by up-regulating odorant-binding proteins genes-OBP-7,10 and OBP-20 expression in the Olfactory System. However, HPX-12 disruption may also cause significant effects on pre-blood meal associated probing abilities, which may be due to fast down-regulation of salivary cocktail proteins such as Anopheline, Apyrase, D7L proteins.

Pathology of Coelomomyces stegomyiae in adult Aedes aegypti ovaries

1988 ◽  
Vol 66 (5) ◽  
pp. 877-884 ◽  
Author(s):  
Christopher J. Lucarotti ◽  
Marina B. Klein
Keyword(s):  
Plasma Membrane ◽  
Aedes Aegypti ◽  
Adult Female ◽  
Blood Meal ◽  
Egg Development ◽  
Yolk Granules ◽  
Receptor Mediated Endocytosis ◽  
Fungal Hyphae ◽  
Tunica Propria ◽  
Coelomomyces Stegomyiae

Coelomomyces stegomyiae (Chytridiomycetes, Blastocladiales) infection in adult female Aedes aegypti (Diptera, Culicidae) is located primarily in the ovaries. Fungal hyphae do not penetrate the germaria or follicles but instead lie between the tunica propria and epithelial sheath within each ovariole and between the epithelial sheath and the peritoneal sheath of the ovary. Aedes aegypti is an anautogenous mosquito requiring a blood meal for egg development; similarly, fungal hyphae in infected ovaries will not differentiate to form resting sporangia until after the mosquito has taken a blood meal. The fungus restricts receptor-mediated endocytosis of vitellogenin by the plasma membrane of the oocyte so that few, if any, vitellin yolk granules form. Thick-walled resting sporangia have formed 72 h after the blood meal has been taken and these will be oviposited by the females in place of the aborted eggs.

LONGEVITY OF BLACK FLIES IN CAPTIVITY

1953 ◽  
Vol 31 (3) ◽  
pp. 304-312 ◽  
Author(s):  
Douglas M. Davies
Keyword(s):  
Adult Female ◽  
Blood Meal ◽  
Blood Parasite ◽  
Black Flies ◽  
Warm Weather ◽  
In Captivity

Adult female black flies were maintained in captivity for as long as 63 days, 34 living for at least 20 days. The flies were kept in longevity tubes in an unhealed room during the spring and summer, and survived longest when sugar was provided as granules and water by a wick. Flies of undetermined age, netted away from the stream, outlived those that emerged one day before the experiment. Females lived longer than males. Flies that partook of a partial blood meal were more viable than those that partook of a full meal. Flies that fed on ducks highly infected with the blood parasite, Leucocytozoon, were less viable than those fed on uninfected ducks, but in most cases flies, collected in the woods, that were not made to feed on ducks outlived those that fed on ducks. On the average, mortality increased during warm weather and decreased during cooler weather.

scholarly journals Immunogenity of Protein Extract from Salivary Gland of Anopheles aconitus in Malaria Endemic Area

2017 ◽  
Vol 18 (1) ◽  
pp. 25
Author(s):  
Mahful Septiawan ◽  
Budayatin Budayatin ◽  
Hidayat Teguh Wiyono ◽  
Kartika Senjarini
Keyword(s):  
Salivary Gland ◽  
Endemic Area ◽  
Protein Profile ◽  
Pathogen Transmission ◽  
Dot Blot ◽  
Protein Extract ◽  
Salivary Gland Extract ◽  
Anopheles Mosquitoes ◽  
Sds Page ◽  
Human Sera

Although malaria had ever been virtually eradicated from Indonesia but currently malaria is recognized as a serious re-emerging threat to public health. This disease is caused by malaria parasite which is transmitted to human host by Anopheles mosquitoes as main vector. It has been widely observed that saliva of mosquito that transmits disease contains several factors that could enhance pathogen infection. Therefore, it should be possible to control pathogen transmission by vaccinating the host against the molecule(s) in saliva that potentiate the infection. However, immunogenic specific component in mosquitoes vectors of Malaria has not yet been identified so far. The objective of this study are to analyze protein profile of SDS-PAGE and to know the immunogity the protein extract of salivary gland from potential vector of Malaria i.e. An. aconitus We used immunogenic reaction between salivary gland extract of these vectors against pool of human sera which were collected from endemic area. The reaction conducted by the dot-blot analyze. SDS-PAGE studies showed 15 major polypeptide bands of 284, 100, 84, 75, 66, 57, 53, 48, 45, 38, 33, 29, 15, 14, and 11 kDa. The dot-blot studies showed that the protein extract of salivary gland from An. aconitus are immunogenic.

scholarly journals Novel CRISPR/Cas9 gene drive constructs inDrosophilareveal insights into mechanisms of resistance allele formation and drive efficiency in genetically diverse populations

2017 ◽  
Author(s):  
Jackson Champer ◽  
Riona Reeves ◽  
Suh Yeon Oh ◽  
Chen Liu ◽  
Jingxian Liu ◽  
...  
Keyword(s):  
Natural Populations ◽  
Model Organism ◽  
Mendelian Inheritance ◽  
Pathogen Transmission ◽  
Resistance Allele ◽  
Gene Drive ◽  
Vector Borne Diseases ◽  
Evolution Of Resistance ◽  
Resistance Rates ◽  
Vector Capacity

ABSTRACTA functioning gene drive system could fundamentally change our strategies for the control of vector-borne diseases by facilitating rapid dissemination of transgenes that prevent pathogen transmission or reduce vector capacity. CRISPR/Cas9 gene drive promises such a mechanism, which works by converting cells that are heterozygous for the drive construct into homozygotes, thereby enabling super-Mendelian inheritance. Though CRISPR gene drive activity has already been demonstrated, a key obstacle for current systems is their propensity to generate resistance alleles. In this study, we developed two CRISPR gene drive constructs based on thenanosandvasapromoters that allowed us to illuminate the different mechanisms by which resistance alleles are formed in the model organismDrosophila melanogaster.We observed resistance allele formation at high rates both prior to fertilization in the germline and post-fertilization in the embryo due to maternally deposited Cas9. Assessment of drive activity in genetically diverse backgrounds further revealed substantial differences in conversion efficiency and resistance rates. Our results demonstrate that the evolution of resistance will likely impose a severe limitation to the effectiveness of current CRISPR gene drive approaches, especially when applied to diverse natural populations.

scholarly journals Regulation of midgut cell proliferation impacts Aedes aegypti susceptibility to dengue virus

2018 ◽  
Author(s):  
Mabel L. Taracena ◽  
Vanessa Bottino-Rojas ◽  
Octavio A.C. Talyuli ◽  
Ana Beatriz Walter-Nuno ◽  
José Henrique M. Oliveira ◽  
...  
Keyword(s):  
Virus Infection ◽  
Aedes Aegypti ◽  
Dengue Virus ◽  
Denv Infection ◽  
Pathogen Transmission ◽  
Intestinal Lumen ◽  
Midgut Cell ◽  
Vector Borne Diseases ◽  
Cellular Processes ◽  
Vector Borne

AbstractAedes aegypti is the vector of some of the most important vector-borne diseases like Dengue, Chikungunya, Zika and Yellow fever, affecting millions of people worldwide. The cellular processes that follow a blood meal in the mosquito midgut are directly associated with pathogen transmission. We studied the homeostatic response of the midgut against oxidative stress, as well as bacterial and dengue virus (DENV) infections, focusing on the proliferative ability of the intestinal stem cells (ISC). Inhibition of the peritrophic matrix (PM) formation led to an increase in ROS production by the epithelial cells in response to contact with the resident microbiota, suggesting that maintenance of low levels of ROS in the intestinal lumen is key to keep ISCs division in balance. We show that dengue virus infection induces midgut cell division in both DENV susceptible (Rockefeller) and refractory (Orlando) mosquito strains. However, the susceptible strain delays the activation of the regeneration process compared with the refractory strain. Impairment of the Delta/Notch signaling, by silencing the Notch ligand Delta using RNAi, significantly increased the susceptibility of the refractory strains to DENV infection of the midgut. We propose that this cell replenishment is essential to control viral infection in the mosquito. Our study demonstrates that the intestinal epithelium of the blood fed mosquito is able to respond and defend against different challenges, including virus infection. In addition, we provide unprecedented evidence that the activation of a cellular regenerative program in the midgut is important for the determination of the mosquito vectorial competence.

scholarly journals The salivary catechol oxidase/peroxidase activities of the mosquito Anopheles albimanus

1993 ◽  
Vol 179 (1) ◽  
pp. 273-287
Author(s):  
J. M. Ribeiro ◽  
R. H. Nussenzveig
Keyword(s):  
Salivary Gland ◽  
Salivary Glands ◽  
Adult Female ◽  
Peroxidase Activity ◽  
Blood Feeding ◽  
Biologically Active ◽  
Relative Molecular Mass ◽  
Catechol Oxidase ◽  
Anopheles Albimanus ◽  
Tissue Destruction

Salivary gland homogenates from adult female Anopheles albimanus mosquitoes relaxed aortic rings preconstricted with noradrenaline (NA). This relaxation is slow and is due to destruction of NA. Incubation of NA with the homogenate yielded a product with a spectrum consistent with the corresponding adrenochrome. Oxidation of NA was enhanced by a superoxide generation system and inhibited by the combined action of superoxide dismutase and catalase. Additionally, peroxidase activity on both synthetic (o-dianisidine) and biologically active (serotonin) substrates was also present in the salivary gland homogenates, this latter activity requiring hydrogen peroxide. Noradrenaline oxidation, serotonin and o-dianisidine peroxidation and vasodilation all co-elute with a heme protein of relative molecular mass 50,000, as determined by molecular sieving chromatography. Peroxidase activity was localized in the posterior (female-specific) lobes of salivary glands and was also detected in nitrocellulose membranes probed by hungry mosquitoes. Protein and peroxidase activities were significantly lower in salivary glands of mosquitoes after probing and feeding on blood. It is suggested that adult female Anopheles albimanus mosquitoes contain a salivary heme peroxidase that functions during blood finding and blood feeding by destroying hemostatically active biogenic amines released by the vertebrate host during tissue destruction.

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