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Dissecting the organ specificity of insecticide resistance candidate genes in Anopheles gambiae : known and novel candidate genes
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Ingham, Victoria A., Jones, Christopher M., Pignatelli, Patricia, Balabanidou, Vasileia, Vontas, John, Wagstaff, Simon C., Moore, Jonathan D. and Ranson, Hilary (2014) Dissecting the organ specificity of insecticide resistance candidate genes in Anopheles gambiae : known and novel candidate genes. BMC Genomics, Volume 15 (Number 1). Article number 1018. doi:10.1186/1471-2164-15-1018 ISSN 1471-2164.
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Official URL: http://dx.doi.org/10.1186/1471-2164-15-1018
Abstract
Background
The elevated expression of enzymes with insecticide metabolism activity can lead to high levels of insecticide resistance in the malaria vector, Anopheles gambiae. In this study, adult female mosquitoes from an insecticide susceptible and resistant strain were dissected into four different body parts. RNA from each of these samples was used in microarray analysis to determine the enrichment patterns of the key detoxification gene families within the mosquito and to identify additional candidate insecticide resistance genes that may have been overlooked in previous experiments on whole organisms.
Results
A general enrichment in the transcription of genes from the four major detoxification gene families (carboxylesterases, glutathione transferases, UDP glucornyltransferases and cytochrome P450s) was observed in the midgut and malpighian tubules. Yet the subset of P450 genes that have previously been implicated in insecticide resistance in An gambiae, show a surprisingly varied profile of tissue enrichment, confirmed by qPCR and, for three candidates, by immunostaining. A stringent selection process was used to define a list of 105 genes that are significantly (p ≤0.001) over expressed in body parts from the resistant versus susceptible strain. Over half of these, including all the cytochrome P450s on this list, were identified in previous whole organism comparisons between the strains, but several new candidates were detected, notably from comparisons of the transcriptomes from dissected abdomen integuments.
Conclusions
The use of RNA extracted from the whole organism to identify candidate insecticide resistance genes has a risk of missing candidates if key genes responsible for the phenotype have restricted expression within the body and/or are over expression only in certain tissues. However, as transcription of genes implicated in metabolic resistance to insecticides is not enriched in any one single organ, comparison of the transcriptome of individual dissected body parts cannot be recommended as a preferred means to identify new candidate insecticide resistant genes. Instead the rich data set on in vivo sites of transcription should be consulted when designing follow up qPCR validation steps, or for screening known candidates in field populations.
Item Type: | Journal Article | ||||||||
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Subjects: | Q Science > QR Microbiology | ||||||||
Divisions: | Faculty of Science, Engineering and Medicine > Research Centres > Warwick Systems Biology Centre | ||||||||
Library of Congress Subject Headings (LCSH): | Anopheles gambiae -- Insecticide resistance, Insecticide resistance -- Genetic aspects | ||||||||
Journal or Publication Title: | BMC Genomics | ||||||||
Publisher: | BioMed Central Ltd. | ||||||||
ISSN: | 1471-2164 | ||||||||
Official Date: | 25 November 2014 | ||||||||
Dates: |
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Volume: | Volume 15 | ||||||||
Number: | Number 1 | ||||||||
Article Number: | Article number 1018 | ||||||||
DOI: | 10.1186/1471-2164-15-1018 | ||||||||
Status: | Peer Reviewed | ||||||||
Publication Status: | Published | ||||||||
Access rights to Published version: | Open Access (Creative Commons) | ||||||||
Date of first compliant deposit: | 28 December 2015 | ||||||||
Date of first compliant Open Access: | 28 December 2015 | ||||||||
Funder: | Seventh Framework Programme (European Commission) (FP7) | ||||||||
Grant number: | 265660 (FP7) | ||||||||
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