Dissecting the organ specificity of insecticide resistance candidate genes in Anopheles gambiae: known and novel candidate genes

Ingham, Victoria ORCID: https://orcid.org/0000-0001-5708-4741, Jones, Christopher ORCID: https://orcid.org/0000-0002-6504-6224, Pignatelli, Patricia, Balabanidou, Vasileia, Vontas, John, Wagstaff, Simon ORCID: https://orcid.org/0000-0003-0577-5537, Moore, Jonathan D and Ranson, Hilary ORCID: https://orcid.org/0000-0003-2332-8247 (2014) 'Dissecting the organ specificity of insecticide resistance candidate genes in Anopheles gambiae: known and novel candidate genes'. BMC Genomics, Vol 15, Issue 1, p. 1018.

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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:	Article
Subjects:	QU Biochemistry > Genetics > QU 470 Genetic structures QX Parasitology > Insects. Other Parasites > QX 515 Anopheles QX Parasitology > Insects. Other Parasites > QX 600 Insect control. Tick control WA Public Health > Preventive Medicine > WA 240 Disinfection. Disinfestation. Pesticides (including diseases caused by)
Faculty: Department:	Biological Sciences > Department of Tropical Disease Biology Biological Sciences > Vector Biology Department
Digital Object Identifer (DOI):	https://doi.org/10.1186/1471-2164-15-1018
Depositing User:	Carmel Bates
Date Deposited:	10 Dec 2014 17:07
Last Modified:	07 Jun 2022 11:09
URI:	https://archive.lstmed.ac.uk/id/eprint/4611

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