Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Sci Rep
2015 Jan 15;5:7795. doi: 10.1038/srep07795.
Show Gene links
Show Anatomy links
Functional characterization of aquaporins and aquaglyceroporins of the yellow fever mosquito, Aedes aegypti.
Drake LL, Rodriguez SD, Hansen IA.
???displayArticle.abstract???
After taking vertebrate blood, female mosquitoes quickly shed excess water and ions while retaining and concentrating the mostly proteinaceous nutrients. Aquaporins (AQPs) are an evolutionary conserved family of membrane transporter proteins that regulate the flow of water and in some cases glycerol and other small molecules across cellular membranes. In a previous study, we found six putative AQP genes in the genome of the yellow fever mosquito, Ae. aegypti, and demonstrated the involvement of three of them in the blood meal-induced diuresis. Here we characterized AQP expression in different tissues before and after a blood meal, explored the substrate specificity of AQPs expressed in the Malpighian tubules and performed RNAi-mediated knockdown and tested for changes in mosquito desiccation resistance. We found that AQPs are generally down-regulated 24 hrs after a blood meal. Ae. aegypti AQP 1 strictly transports water, AQP 2 and 5 demonstrate limited solute transport, but primarily function as water transporters. AQP 4 is an aquaglyceroporin with multiple substrates. Knockdown of AQPs expressed in the MTs increased survival of Ae. aegypti under dry conditions. We conclude that Malpighian tubules of adult female yellow fever mosquitoes utilize three distinct AQPs and one aquaglyceroporin in their osmoregulatory functions.
Figure 1. Aedes aegypti AQP expression in various parts of the alimentary canal.Expression was assayed using qPCR. The data represent relative quantification of Aedes AQPs which were normalized by qPCR analysis of ribosomal protein S7 (rpS7) mRNA levels in the cDNA samples. Values are means ± S.E. (error bars) of triplicate biological samples. The Y-axis shows expression ratios in arbitrary units with the lowest expressing tissue set as 1. Means separated by Tukey–Kramer HSD (p < 0.05). Means which share the same letter are not significantly different. RNA was isolated from organs/body parts of adult female mosquitoes unfed (light shaded columns) and 24 hrs after a blood meal (dark shaded columns). C-crop, FG-foregut, MG-midgut, HG-hindgut, R-rectum, MT – Malpighian tubules, OV – ovaries.
Figure 2. Water permeability analysis of Ae.
aegypti AQPs expressed in Xenopus
laevis oocytes.(a) Day 3 post-injection with AQP cRNA, Western blot analyses of oocyte lysates using Myc-tag antibody (the AQP tetramer band is shown). The blots were cropped, and the full-length blots are presented in the supplementary information (see Supplementary Fig. S1). (b) Effect of heterologous expression of Aedes AQPs on water permeability of Xenopus oocytes. The light-shaded columns represent oocytes that were pre- treated with HgCl2. Each AQP group n = 9, and their mean separated by Tukey–Kramer HSD (p < 0.05). Means which share the same letter are not significantly different.
Figure 3. Various solute transport capabilities of Ae.
aegypti AQPs expressed in Xenopus laevis oocytes.Solute uptake was reported as an oocyte swelling rate [d(V/V0)/dt] and measured in μm/sec. There were six solutes tested, in each AQP group n = 5. The molecular structure of each solute is indicated above each graph. The means were separated by the Tukey-Kramer HSD (p < 0.05). Means which share the same letter are not significantly different.
Figure 4. RNAi-mediated knockdown of Ae.
aegypti AQPs demonstrate desiccation resistance.Effect of RNAi-mediated AQP knockdown on Aedes survival under desiccation conditions. AQP 1, 4 and 5 knockdown mosquitoes have a significant increase in survival compared to the eGFP control group. Kaplan-Meier and log-rank tests were performed using Graphpad Prism software.
Agre,
Aquaporin water channels--from atomic structure to clinical medicine.
2002, Pubmed
Agre,
Aquaporin water channels--from atomic structure to clinical medicine.
2002,
Pubmed Becker,
The regulation of trehalose metabolism in insects.
1996,
Pubmed Benoit,
Aquaporins are critical for provision of water during lactation and intrauterine progeny hydration to maintain tsetse fly reproductive success.
2014,
Pubmed Bradley,
Physiology of osmoregulation in mosquitoes.
1987,
Pubmed Carpenter,
SLC7 amino acid transporters of the yellow fever mosquito Aedes aegypti and their role in fat body TOR signaling and reproduction.
2012,
Pubmed Drake,
The Aquaporin gene family of the yellow fever mosquito, Aedes aegypti.
2010,
Pubmed Drake,
RNAi-mediated gene knockdown and in vivo diuresis assay in adult female Aedes aegypti mosquitoes.
2012,
Pubmed Duchesne,
Mosquito (Aedes aegypti ) aquaporin, present in tracheolar cells, transports water, not glycerol, and forms orthogonal arrays in Xenopus oocyte membranes.
2003,
Pubmed
,
Xenbase Esquivel,
Transcriptomic evidence for a dramatic functional transition of the malpighian tubules after a blood meal in the Asian tiger mosquito Aedes albopictus.
2014,
Pubmed Gonen,
The structure of aquaporins.
2006,
Pubmed
,
Xenbase Hansen,
AaCAT1 of the yellow fever mosquito, Aedes aegypti: a novel histidine-specific amino acid transporter from the SLC7 family.
2011,
Pubmed
,
Xenbase Hays,
A novel protein produced by the vitellogenic fat body and accumulated in mosquito oocytes.
1990,
Pubmed Ikeguchi,
Water transport in aquaporins: molecular dynamics simulations.
2009,
Pubmed Jensen,
The mechanism of glycerol conduction in aquaglyceroporins.
2001,
Pubmed Jung,
Molecular structure of the water channel through aquaporin CHIP. The hourglass model.
1994,
Pubmed
,
Xenbase Jung,
Molecular characterization of an aquaporin cDNA from brain: candidate osmoreceptor and regulator of water balance.
1994,
Pubmed
,
Xenbase Kikawada,
Trehalose transporter 1, a facilitated and high-capacity trehalose transporter, allows exogenous trehalose uptake into cells.
2007,
Pubmed
,
Xenbase King,
From structure to disease: the evolving tale of aquaporin biology.
2004,
Pubmed Leparc-Goffart,
Chikungunya in the Americas.
2014,
Pubmed Li,
Plant aquaporins: roles in plant physiology.
2014,
Pubmed Liu,
Aquaporin water channel AgAQP1 in the malaria vector mosquito Anopheles gambiae during blood feeding and humidity adaptation.
2011,
Pubmed
,
Xenbase Liu,
Impact of trehalose transporter knockdown on Anopheles gambiae stress adaptation and susceptibility to Plasmodium falciparum infection.
2013,
Pubmed Marchette,
Isolation of Zika virus from Aedes aegypti mosquitoes in Malaysia.
1969,
Pubmed Musa-Aziz,
Using fluorometry and ion-sensitive microelectrodes to study the functional expression of heterologously-expressed ion channels and transporters in Xenopus oocytes.
2010,
Pubmed
,
Xenbase Philip,
Aquaporins play a role in desiccation and freeze tolerance in larvae of the goldenrod gall fly, Eurosta solidaginis.
2008,
Pubmed Pietrantonio,
Cloning of an aquaporin-like cDNA and in situ hybridization in adults of the mosquito Aedes aegypti (Diptera: Culicidae).
2000,
Pubmed Rao,
Similarity of the product of the Drosophila neurogenic gene big brain to transmembrane channel proteins.
1990,
Pubmed Robert Michaud,
Metabolomics reveals unique and shared metabolic changes in response to heat shock, freezing and desiccation in the Antarctic midge, Belgica antarctica.
2008,
Pubmed Spring,
The role of aquaporins in excretion in insects.
2009,
Pubmed Staniscuaski,
Expression analysis and molecular characterization of aquaporins in Rhodnius prolixus.
2013,
Pubmed Tatsumi,
Drosophila big brain does not act as a water channel, but mediates cell adhesion.
2009,
Pubmed Thompson,
Glucogenic blood sugar formation in an insect Manduca sexta L.: asymmetric synthesis of trehalose from 13C enriched pyruvate.
2003,
Pubmed Törnroth-Horsefield,
Structural insights into eukaryotic aquaporin regulation.
2010,
Pubmed Tsujimoto,
Organ-specific splice variants of aquaporin water channel AgAQP1 in the malaria vector Anopheles gambiae.
2013,
Pubmed Verkman,
Aquaporins in clinical medicine.
2012,
Pubmed Verkman,
Aquaporins: important but elusive drug targets.
2014,
Pubmed Wallace,
Distinct transport selectivity of two structural subclasses of the nodulin-like intrinsic protein family of plant aquaglyceroporin channels.
2005,
Pubmed
,
Xenbase Yoder,
Stress-induced accumulation of glycerol in the flesh fly, Sarcophaga bullata: evidence indicating anti-desiccant and cryoprotectant functions of this polyol and a role for the brain in coordinating the response.
2006,
Pubmed
