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- Bonaventure Omondi, Aman, et al.
(författare)
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Age-dependent regulation of host seeking in Anopheles coluzzii
- 2019
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Ingår i: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322. ; 9
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Tidskriftsartikel (refereegranskat)abstract
- Behavioural attraction of the malaria vector Anopheles coluzzii to human host odour increases during adult maturation. We have previously demonstrated that the onset of host seeking in An. coluzzii coincides with an increased sensitivity of the CO2-sensitive neurons and abundance of chemosensory receptor gene transcripts in the maxillary palp. In this study, we extend our analysis to the antenna. Functional characterisation of the near-complete repertoire of odorant receptors (Ors) expressed in this tissue, to fractioned human odour, reveals a subset of salient human odorants to be detected by Ors at physiological relevant concentrations. When presented as a blend in their ratio of natural emission, these odorants elicit attraction by host-seeking mosquitoes, emphasising that Ors alone can mediate this behaviour. However, the same blend inhibits attraction in teneral mosquitoes. This switch in behavioural response indicates a change in valence during adult maturation. Quantitative analysis of Or transcript abundance and in vivo electrophysiological analysis reveal that the olfactory system of female An. coluzzii undergoes concerted changes that correlate with the onset of host seeking. We conclude that changes in Or abundance modulate peripheral olfactory coding, resulting in ecologically relevant behavioural effects.
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| 3. |
- Bonaventure Omondi, Aman, et al.
(författare)
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Evaluation of reference genes for insect olfaction studies
- 2015
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Ingår i: Parasites and Vectors. - : Springer Science and Business Media LLC. - 1756-3305. ; 8
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Tidskriftsartikel (refereegranskat)abstract
- Background: Quantitative reverse transcription PCR (qRT-PCR) is a robust and accessible method to assay gene expression and to infer gene regulation. Being a chain of procedures, this technique is subject to systematic error due to biological and technical limitations mainly set by the starting material and downstream procedures. Thus, rigorous data normalization is critical to grant reliability and repeatability of gene expression quantification by qRT-PCR. A number of 'housekeeping genes', involved in basic cellular functions, have been commonly used as internal controls for this normalization process. However, these genes could themselves be regulated and must therefore be tested a priori.Methods: We evaluated eight potential reference genes for their stability as internal controls for RT-qPCR studies of olfactory gene expression in the antennae of Rhodnius prolixus, a Chagas disease vector. The set of genes included were: a-tubulin; beta-actin; Glyceraldehyde-3-phosphate dehydrogenase; Eukaryotic initiation factor 1A; Glutathione-S-transferase; Serine protease; Succinate dehydrogenase; and Glucose-6-phosphate dehydrogenase. Five experimental conditions, including changes in age, developmental stage and feeding status were tested in both sexes.Results: We show that the evaluation of candidate reference genes is necessary for each combination of sex, tissue and physiological condition analyzed in order to avoid inconsistent results and conclusions. Although, Normfinder and geNorm software yielded different results between males and females, five genes (SDH, Tub, GAPDH, Act and G6PDH) appeared in the first positions in all rankings obtained. By using gene expression data of a single olfactory coreceptor gene as an example, we demonstrated the extent of changes expected using different internal standards.Conclusions: This work underlines the need for a rigorous selection of internal standards to grant the reliability of normalization processes in qRT-PCR studies. Furthermore, we show that particular physiological or developmental conditions require independent evaluation of a diverse set of potential reference genes.
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| 4. |
- Bonaventure Omondi, Aman, et al.
(författare)
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Functional development of carbon dioxide detection in the maxillary palp of Anopheles gambiae
- 2015
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Ingår i: Journal of Experimental Biology. - : The Company of Biologists. - 0022-0949 .- 1477-9145. ; 218, s. 2482-2488
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Tidskriftsartikel (refereegranskat)abstract
- Olfactory information drives several behaviours critical for the survival and persistence of insect pests and vectors. Insect behaviour is variable, linked to their biological needs, and regulated by physiological dynamics. For mosquitoes, CO2 is an important cue that signifies the presence of a host, and which elicits activation and attraction. To investigate the genetic basis of olfactory modulation in mosquitoes, we assayed changes in CO2 detection from receptor gene expression through physiological function to behaviour, associated with the onset of host seeking in the malaria vector, Anopheles gambiae. The gene encoding a subunit of the CO2 receptor, AgGr22, was found to be significantly up-regulated in host-seeking females, consistent with a significant increase in sensitivity of CO2-responsive neurons (cpA) housed in capitate peg sensilla of the maxillary palp. In addition, the odorant receptor AgOr28, which is expressed in cpC neurons, was significantly up-regulated. In contrast, AgOr8, which is expressed in cpB neurons, was not affected by this change in physiological state, in agreement with results for the obligate co-receptor Orco. Moreover, the sensitivity of the cpB neuron to (R)-1-octen-3-ol, a well-known mammalian kairomone, did not change in response to the onset of host seeking. The concentration of CO2 flux influenced both the propensity of A. gambiae to take off into the wind and the speed with which this activation occurred. Female A. gambiae mosquitoes responded to CO2 whether mature for host seeking or not, but onset of host seeking enhanced sensitivity and speed of activation at relevant doses of CO2.
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| 5. |
- Bonaventure Omondi, Aman, et al.
(författare)
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Molecular basis of peripheral olfactory plasticity in Rhodnius prolixus, a Chagas disease vector
- 2015
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Ingår i: Frontiers in Ecology and Evolution. - : Frontiers Media SA. - 2296-701X. ; 3
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Tidskriftsartikel (refereegranskat)abstract
- Olfaction is fundamental for most animals and critical for different aspects of triatomine biology, including host-seeking, reproduction, avoidance of predators, and aggregation in shelters. Ethological and physiological aspects of these olfactory-mediated behaviors are well-understood, but their molecular bases are still largely unknown. Here we investigated changes in the molecular mechanisms at the peripheral olfactory level in response to different physiological and developmental conditions. For this, the antennal expression levels of the odorant (Orco) and ionotropic (IR8a,IR25a, andIR76b) coreceptor genes were determined inRhodnius prolixusby means of quantitative real-time PCR (qRT-PCR) analysis. Gene expression changes were analyzed to test the effect of feeding and imaginal molt for both sexes. Moreover, we analyzed whether expression of these genes changed during the early life of adult bugs. Under these conditions bugs display distinct behavioral responses to diverse chemical stimuli. A significantly decreased expression was induced by blood feeding on all coreceptor genes. The expression of all genes was significantly increased following the imaginal molt. These results show that olfactory coreceptor genes have their expression altered as a response to physiological or developmental changes. Our study suggests that olfactory coreceptor genes confer adaptability to the peripheral olfactory function, probably underlying the known plasticity of triatomine olfactory-mediated behavior. Read Full Text
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| 6. |
- Christ, Peter, et al.
(författare)
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Feeding-induced changes in allatostatin-A and short neuropeptide F in the antennal lobes affect odor-mediated host seeking in the yellow fever mosquito, Aedes aegypti
- 2017
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Ingår i: PLoS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 12
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Tidskriftsartikel (refereegranskat)abstract
- Aedes aegypti is a model species in which the endogenous regulation of odor-mediated host seeking behavior has received some attention. Sugar feeding and host seeking in female A. aegypti are transiently inhibited following a blood meal. This inhibition is partially mediated by short neuropeptide F (sNPF). The paired antennal lobes (ALs), as the first processing centers for olfactory information, has been shown to play a significant role in the neuropeptidergic regulation of odor-mediated behaviors in insects. The expression of sNPF, along with other peptides in the ALs of A. aegypti, indicate parallel neuromodulatory systems that may affect olfactory processing. To identify neuropeptides involved in regulating the odor-mediated host seeking behavior in A. aegypti, we use a semi-quantitative neuropeptidomic analysis of single ALs to analyze changes in the levels of five individual neuropeptides in response to different feeding regimes. Our results show that the level of sNPF-2, allatostatin-A-5 (AstA-5) and neuropeptide-like precursor-1-5 (NPLP-1-5), but not of tachykinin- related-peptides and SIFamide (SIFa), in the AL of female mosquitoes, changes 24 h and 48 h post-blood meal, and are dependent on prior access to sugar. To assess the role of these neuropeptides in modulating host seeking behavior, when systemically injected individually, sNPF-2 and AstA-5 significantly reduced host seeking behavior. However, only the injection of the binary mixture of the two neuropeptides lead to a host seeking inhibition similar to that observed in blood fed females. We conclude that modulation of the odor mediated host seeking behavior of A. aegypti is likely regulated by a dual neuropeptidergic pathway acting in concert in the ALs.
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| 7. |
- Christ, Peter, et al.
(författare)
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Functional characterization of mosquito short neuropeptide F receptors
- 2018
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Ingår i: Peptides. - : Elsevier BV. - 0196-9781 .- 1873-5169. ; 103, s. 31-39
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Tidskriftsartikel (refereegranskat)abstract
- Mosquito blood feeding transiently inhibits sugar-and host seeking through neuropeptide signaling. Short neuropeptide F (sNPF) is one of the neuromodulators involved in this regulation. Here, we identified the genes for the sNPF precursor and the sNPF receptor in the southern house mosquito, Culex quinquefasciatus. Comparative analyses are made with the genes of the sNPF precursor and receptor from two other important vectors, Aedes aegypti and Anopheles coluzzii. We functionally characterized the receptors in all three species using endogenous neuropeptides, and quantified their transcript expression following a blood meal and a sugar meal. Our analysis reveals several Cx. quinquefasciatus-specific duplications of the sNPF-3 isoform on the sNPF precursor, which are not reflected in the precursors of the other two species. In contrast, the structure of the sNPF receptors is highly conserved within mosquitoes, and a putative ligand binding region is proposed and discussed. Reflecting the high structural conservation, the sNPF receptor sensitivity to endogenous sNPF iso-forms is conserved across mosquito species. Using quantitative real time PCR, we demonstrate that transcript abundance of the sNPF receptor and precursor is regulated following feeding, only in Cx. quinquefasciatus. We discuss our findings in relation to previous work on sNPF signaling and its role in feeding regulation.
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| 8. |
- Christ, Peter, et al.
(författare)
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Functional characterization of the dual allatostatin-A receptors in mosquitoes
- 2018
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Ingår i: Peptides. - : Elsevier BV. - 0196-9781 .- 1873-5169. ; 99, s. 44-55
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Tidskriftsartikel (refereegranskat)abstract
- The neuropeptide allatostatin-A (AstA) and its cognate receptors (AstARs) are involved in the modulation of feeding behavior, which in hematophagous insects includes the regulation of the disease vector-related behaviors, host seeking and blood feeding. In mosquitoes and other dipterans, there are two copies of AstAR, contrasting with the single copy found in other insects. In this study, we identified and cloned the dual AstAR system of two important disease vectors Aedes aegypti and Culex quinquefasciatus, and compared them with those previously described, including those in Anopheles coluzzii and Drosophila melanogaster. Phylogenetic analysis of the AstARs revealed that the mosquito AstAR1s has retained a similar amino acid sequence as the AstARs from non-dipteran insect species. Intron analysis revealed that the number of introns accumulated in the AstAR2s is similar to that in other insects, and that introns are conserved within the receptor types, but that only the final two introns are conserved across AstAR1s and 2s. We functionally characterized the dual AstARs in An. coluzzii, Ae. aegypti and Cx. quinquefasciatus by stably expressing the receptors in a Chinese hamster oocyte cell line (CHO) also stably expressing a promiscuous G-protein (G16), and challenged them with the endogenous isoforms of AstA from the three mosquito species. In the culicine mosquitoes, Ae. aegypti and Cx. quinquefasciatus, the AstARs demonstrated differential sensitivity to AstA, with the AstAR2s displaying a higher sensitivity than the AstAR1s, suggesting a divergence of functional roles for these AstARs. In contrast, both An. coluzzii AstARs demonstrated a similar sensitivity to the AstA ligands. We discuss our findings in the light of AstA acting as a regulator of blood feeding in mosquitoes. A better understanding of the regulation of host seeking and blood feeding in vector mosquitoes will lead to the rational development of novel approaches for vector control.
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| 9. |
- Claesson, Rickard, et al.
(författare)
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HbA1c as a predictor of diabetes after gestational diabetes mellitus
- 2017
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Ingår i: Primary Care Diabetes. - : Elsevier BV. - 1751-9918. ; 11:1, s. 46-51
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Tidskriftsartikel (refereegranskat)abstract
- Aim We wanted to investigate third-trimester HbA1c as a predictor of diabetes after gestational diabetes mellitus (GDM). Methods Women with GDM were followed up prospectively for five years from pregnancy to detect the development of diabetes. The ability of HbA1c to predict diabetes was evaluated with receiver-operating characteristic (ROC) curves and logistic regression analysis. Results By five years, 73 of 196 women had been diagnosed with diabetes. An optimal cut-off point for HbA1c of 36 mmol/mol (5.4%) could predict diabetes with 45% sensitivity and 92% specificity. For HbA1c ≥39 mmol/mol (≥5.7%), sensitivity, specificity, and positive predictive value were 30%, 97%, and 91%, respectively. In logistic regression analysis, adjusting for the diagnostic glucose concentration during pregnancy, HbA1c levels in the upper quartile (≥36 mmol/mol) were associated with a 5.5-fold increased risk of diabetes. Conclusion Third-trimester HbA1c levels in the pre-diabetes range revealed women with post-partum diabetes with high specificity and high positive predictive value. HbA1c testing could be used as a strategy to select high-risk women for lifestyle interventions aimed at prevention of diabetes starting during pregnancy. The results should encourage further validation in other populations using new diagnostic criteria for GDM.
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