Biomedical subjects

T Guillemaud

Publications and source records attributed to T Guillemaud.

9 recordsLinked to original sources

PubMed1998

An overview of the evolution of overproduced esterases in the mosquito Culex pipiens.

Insecticide resistance genes have developed in a wide variety of insects in response to heavy chemical application. Few of these examples of adaptation in response to rapid environmental change have been studied both at the population level and at the gene level. One of these is the evolution of the overproduced esterases that are involved in resistance to organophosphate insecticides in the mosquito Culex pipiens. At the gene level, two genetic mechanisms are involved in esterase overproduction, namely gene amplification and gene regulation. At the population level, the co-occurrence of the same amplified allele in distinct geographic areas is best explained by the importance of passive transportation at the worldwide scale. The long-term monitoring of a population of mosquitoes in southern France has enabled a detailed study to be made of the evolution of resistance genes on a local scale, and has shown that a resistance gene with a lower cost has replaced a former resistance allele with a higher cost.

Alleles↗

PubMed1998

Evaluating gene flow using selected markers: a case study.

The extent to which an organism is locally adapted in an environmental pocket depends on the selection intensities inside and outside the pocket, on migration, and on the size of the pocket. When two or more loci are involved in this local adaptation, measuring their frequency gradients and their linkage disequilbria allows one to disentangle the forces-migration and selection-acting on the system. We apply this method to the case of a local adaptation to organophosphate insecticides in the mosquito Culex pipiens pipiens in southern France. The study of two different resistance loci allowed us to estimate with support limits gene flow as well as selection pressure on insecticide resistance and the fitness costs associated with each locus. These estimates permit us to pinpoint the conditions for the maintenance of this pocket of adaptation as well as the effect of the interaction between the two resistance loci.

Animals↗

PubMed1998

Polymorphisms and fluctuations in copy number of amplified esterase genes in Culex pipiens mosquitoes.

In Culex pipiens mosquitoes, A2 esterase alleles are co-amplified with B2 esterase alleles in response to selection with organophosphate insecticides. In this study the amplified A2 and B2 sequences were compared between twelve strains from four continents by restriction mapping. The restriction maps were almost identical in each strain throughout 22 kb surrounding the genes, suggesting that this represents a constant core sequence. A polymorphism was found in two strains collected from Egypt and Kenya in the mid 1980s. This polymorphism was present in all copies of the amplicon, which suggests that a mechanism of sequence homogenization was operating, i.e. concerted evolution. These two strains were almost certainly descendants from the same population and migration probably occurred along the River Nile. Although the maps were almost identical in each strain, dot blotting demonstrated that amplification levels differed by up to 13-fold between strains. Thus the presence of the A2-B2 haplotype cannot be used to indicate the level of amplification or any particular degree of resistance.

Animals↗

PubMed1997

Contrasting levels of variability between cytoplasmic genomes and incompatibility types in the mosquito Culex pipiens.

Reproductive incompatibilities called cytoplasmic incompatibilities are known to affect a large number of arthropod species and are mediated by Wolbachia, a maternally transmitted microorganism. The crossing relationships between strains of potential hosts define their incompatibility types and it is generally assumed that differences between strains of Wolbachia induce different crossing types. Among all the described host species, the mosquito, Culex pipiens, displays the greatest variability of cytoplasmic incompatibility crossing types. We analysed mitochondrial and bacterial DNA variability in Culex pipiens in order to investigate some possible causes of incompatibility crossing type variability. We sequenced fragments of the ftsZ gene, and the A + T-rich control region of the mtDNA. We also sequenced the second subunit of the mitochondrial cytochrome oxidase (COII) gene, in Culex pipiens and a closely related species, C. torrentium, in order to verify the usefulness of the A + T-rich region for the present purposes. No variability was found in the Wolbachia ftsZ gene fragment, and very limited variation of the mitochondrial marker whatever the compatibility type or the origin of the host. A low variability was found in the A + T-rich region and comparison of divergence of the A + T-rich region and COII gene between C. pipiens and C. torrentium did not reveal any special constraints affecting this region. In contrast to observations in other host species, variability of incompatibility crossing types is not due to multiple infections by distantly related Wolbachia strains.

Animals↗

PubMed1997

Variation of dominance of newly arisen adaptive genes.

Newly arisen adaptive alleles such as insecticide resistance genes represent a good opportunity to investigate the theories put forth to explain the molecular basis of dominance and its possible evolution. Dominance levels of insecticide resistance conferred by insensitive alleles of the acetylcholinesterase gene were analyzed in five resistant strains of the mosquito Culex pipiens. Dominance levels were found to differ between strains, varying from partial recessivity to complete dominance. This variation was not explained by differences in catalytic properties of the enzyme, since four of the five resistant strains had identical inhibition properties for the insensitive acetylcholinesterase. Among these four laboratory strains and in individuals collected from natural populations, we found a correlation between increased acetylcholinesterase activities and higher dominance levels. We propose a molecular explanation for how variation in acetylcholinesterase activity may result in variation of dominance level. We also conjecture that the four resistant strains did not differ in their amino acid sequence in the catalytically active regions of acetylcholinesterase, but that the expression of the gene was regulated by either neighboring or distant sites, thereby modifying the dominance level. Under this interpretation, dominance levels may evolve in this system, since heritable variation in acetylcholinesterase activity was found.

Acetylcholinesterase↗

PubMed1997

Esterases A5-B5 in organophosphate-resistant Culex pipiens from Italy.

Culex pipiens mosquitos from Lignano city, Udine province, northeast Italy, were found to carry over-produced non-specific esterases A1, A2-B2 and A4-B4 or A5-B5, detected by starch gel electrophoresis, giving multiple resistance to organophosphorus insecticides. In order to differentiate between A4-B4 and A5-B5 esterases, the latter known only from Cyprus whereas the former is widespread in Italy and elsewhere, restriction fragment length polymorphism (RFLP) analysis was performed at the esterase B locus. Both B4 and B5 haplotypes were found. This is the first record of A5-B5 esterase-mediated resistance in continental Europe.

Animals↗

PubMed1996

Testing the unique amplification event and the worldwide migration hypothesis of insecticide resistance genes with sequence data.

In the mosquito Culex pipiens, over-production of esterases is a common insecticide resistance mechanism. Different alleles at the esterase A and B loci are known to occur in natural populations, as shown by enzyme electrophoresis and RFLP studies on the esterase B locus. Here we analyse the variability of the esterase A locus at the nucleotide level in mosquitoes possessing or lacking over-produced esterase A. A surprisingly high level of nucleotide polymorphism is found in coding and noncoding regions, and the extent of polymorphism detected is higher than that previously described for the most polymorphic loci in Drosophila. We also show that eight strains from different localities (Africa, America, Asia) that possess the over-produced esterase A2 share the same nucleotide sequence at the esterase A locus, strongly supporting the evolutionary scenario of a unique event of amplification of this gene followed by dispersal through migration rather than the occurrence of multiple independent mutational events of the gene.

Amino Acid Sequence↗

PubMed1996

Molecular identification of a Wolbachia endosymbiont in a Tetranychus urticae strain (Acari: Tetranychidae).

Wolbachia, a maternally inherited bacterium, is involved in alterations of arthropod sexuality. Reproductive incompatibilities are often observed in miles, but the existence of this microorganism in their cytoplasm has not yet been demonstrated. We identified the presence of Wolbachia in a strain of the spider mite Tetranychus urticae based on the amplification and sequencing of part of the 16S rDNA and ftsZ genes. In order to establish the phylogenetic relationships between Wolbachia found in T. urticae and in other arthropods, we aligned the resulting sequences with already published ones. For both 16S and ftsZ genes the Wolbachia carried by T. urticae clustered together with Wolbachia found in other arthropods.

Animals↗

PubMed1996

Coamplification of esterase A and B genes as a single unit in Culex pipiens mosquitoes.

In Culex pipiens mosquitoes, resistance to organophosphorous insecticides often results from increased detoxification by two types of esterases, A and B, which are closely linked. Overproduction of all esterase B so far investigated (B1, B2, B4, B5 and B6) is from gene amplification. An esterase A gene (esterase A2) has recently been cloned from mosquitoes with the overproduced esterases A2 and B2, and amplification of this gene has also been reported. We describe the cDNA sequences of three additional esterase genes from insecticide-resistant strains of Culex pipiens originating from France and California which show at least 93 per cent homology with the esterase A2 gene sequence. Restriction enzyme mapping shows that the esterase A gene lies within 2.2 kb of the esterase B gene. In mosquitoes with overproduced esterases A2 and B2, the amplification level of esterase A is equal to that of esterase B suggesting that the genes are coamplified. Furthermore, we show that in one strain with an overproduced A esterase (A1), gene amplification cannot account for the increased protein level. This indicates that overproduction of esterases A can be achieved through two different mechanisms: gene amplification and a regulatory mechanism--the nature of which remains to be identified.

Animals↗