Genetic diversity of canine olfactory receptors

<p>Abstract</p> <p>Background</p> <p>Evolution has resulted in large repertoires of olfactory receptor (OR) genes, forming the largest gene families in mammalian genomes. Knowledge of the genetic diversity of olfactory receptors is essential if we are to understand the differences in olfactory sensory capability between individuals. Canine breeds constitute an attractive model system for such investigations.</p> <p>Results</p> <p>We sequenced 109 OR genes considered representative of the whole OR canine repertoire, which consists of more than 800 genes, in a cohort of 48 dogs of six different breeds. SNP frequency showed the overall level of polymorphism to be high. However, the distribution of SNP was highly heterogeneous among OR genes. More than 50% of OR genes were found to harbour a large number of SNP, whereas the rest were devoid of SNP or only slightly polymorphic. Heterogeneity was also observed across breeds, with 25% of the SNP breed-specific. Linkage disequilibrium within OR genes and OR clusters suggested a gene conversion process, consistent with a mean level of polymorphism higher than that observed for introns and intergenic sequences. A large proportion (47%) of SNP induced amino-acid changes and the Ka/Ks ratio calculated for all alleles with a complete ORF indicated a low selective constraint with respect to the high level of redundancy of the olfactory combinatory code and an ongoing pseudogenisation process, which affects dog breeds differently.</p> <p>Conclusion</p> <p>Our demonstration of a high overall level of polymorphism, likely to modify the ligand-binding capacity of receptors distributed differently within the six breeds tested, is the first step towards understanding why Labrador Retrievers and German Shepherd Dogs have a much greater potential for use as sniffer dogs than Pekingese dogs or Greyhounds. Furthermore, the heterogeneity in OR polymorphism observed raises questions as to why, in a context in which most OR genes are highly polymorphic, a subset of these genes is not? This phenomenon may be related to the nature of their ligands and their importance in everyday life.</p

Coat colour in dogs: identification of the Merle locus in the Australian shepherd breed

BACKGROUND: Coat colours in canines have many natural phenotypic variants. Some of the genes and alleles involved also cause genetic developmental defects, which are also observed in humans and mice. We studied the genetic bases of the merle phenotype in dogs to shed light on the pigmentation mechanisms and to identify genes involved in these complex pathways. The merle phenotype includes a lack of eumelanic pigmentation and developmental defects, hearing impairments and microphthalmia. It is similar to that observed in microphthalmia mouse mutants. RESULTS: Taking advantage of the dog as a powerful genetic model and using recently available genomic resources, we investigated the segregation of the merle phenotype in a five-generation pedigree, comprising 96 sampled Australian shepherd dogs. Genetic linkage analysis allowed us to identify a locus for the merle phenotype, spanning 5.5 megabases, at the centromeric tip of canine chromosome 10 (CFA10). This locus was supported by a Lod score of 15.65 at a recombination fraction θ = 0. Linkage analysis in three other breeds revealed that the same region is linked to the merle phenotype. This region, which is orthologous to human chromosome 12 (HSA12 q13-q14), belongs to a conserved ordered segment in the human and mouse genome and comprises several genes potentially involved in pigmentation and development. CONCLUSION: This study has identified the locus for the merle coat colour in dogs to be at the centromeric end of CFA10. Genetic studies on other breeds segregating the merle phenotype should allow the locus to be defined more accurately with the aim of identifying the gene. This work shows the power of the canine system to search for the genetic bases of mammalian pigmentation and developmental pathways

Les réseaux d'approvisionnement [Le domaine de l'économie]

International audienc

Les réseaux d'approvisionnement [Le domaine de l'économie]

International audienc

Les réseaux d'approvisionnement [Le domaine de l'économie]

International audienc

Les réseaux d'approvisionnement [Des activités économiques difficiles à caractériser]

International audienc

Unexpected and spatially structured genetic diversity of the relict population of the endangered corsican land snail Tyrrhenaria ceratina

We thank all the volunteers who realized the sample collection, especially the team of the Centre Permanent d’Initiatives pour l’Environnement d’Ajaccio mobilized for the project “Aio lumacchi, Allez les escargots” funded by Office Français de la Biodiversité and the wardens of the Collectivité Territoriale de Corse. We also thank DREAL Corse for funding support. Handling and sampling of this protected species were performed under permits: Arrêtés préfectoraux 2 A 2018-01-29-001, 2 A 2020-03-25-001 and 2 A-2021-02-25-003.International audiencePopulation genetics of threatened species provides information about evolutionary pressures over those populations and thus may inform conservation management strategies. However, conservation genetics still has a low impact on conservation practices. This study's aim is to integrate genetics in the conservation management of the only-known population of an extremely narrow-range endemic Corsican snail-Tyrrhenaria ceratina-, whose distribution area is restricted to the Ricantu site in Corsica. Using non-invasive DNA samples of 210 individuals, we amplified 13 microsatellites loci to assess the population viability, genetic structure and demographic history of the population, along with the estimation of the historical and contemporary gene flow between identified genetic clusters. We also estimated the dispersal ability of the species. Our results showed a surprisingly high genetic diversity, along with a pattern of isolation by distance (IBD) and a strongly spatialized genetic structure. Furthermore, we underlined a low functional connectivity, along with evidence of a recent decline in the population size, which are both likely due to a historical fragmentation between the sampled areas, caused by anthropization. Overall, this study allows to provide a first insight about the functioning of the population, to guide future conservation actions for the species

Mechanism of action of sprG1-encoded type I toxins in Staphylococcus aureus: from membrane alterations to mesosome-like structures formation and bacterial lysis

International audiencesprG1/ SprF1 is a type I toxin-antitoxin system located on Staphylococcus aureus prophage. It has previously been shown that the two toxins, SprG1 31 and SprG1 44 , encoded by the sprG1 gene, are two membrane-associated peptides structured in a single α-helix. Overexpression of these two peptides leads to growth inhibition and even S. aureus death. In this study, we investigated the involvement of each peptide in this toxicity, the sequence requirements necessary for SprG1 31 toxicity, and the mechanism of action of these two peptides. Our findings show that both peptides, when expressed individually, are able to stop growth, with higher toxicity observed for SprG1 31 . The combination of a hydrophobic domain and a charged domain located only at the C-terminus is necessary for this toxicity, likely to retain the orientation of the transmembrane domain. A net cationic charge for SprG1 31 is not essential to induce a growth defect in S. aureus . Furthermore, we established a chronology of toxic events following overexpression to gain insights into the mode of action of SprG1 44 and SprG1 31 . We demonstrated that mesosome-like structures are already formed when membrane is depolarized, about 20 min after peptides induction. This membrane depolarization occurs concomitantly with a depletion of intracellular ATP, leading to S. aureus growth arrest. Moreover, we hypothesized that SprG1 44 and SprG1 31 do not form large pores in the S. aureus membrane, as ATP is not excreted into the extracellular medium, and membrane permeabilization is delayed relative to membrane depolarization. The next challenge is to identify the conditions under which SprG1 44 and SprG1 31 are naturally expressed, and to uncover their potential roles during staphylococcal growth, colonization, and infection

A multiscale analysis of landscape resistance reveals genetic isolates in an endangered forest-specialist species the Barbary macaque (Macaca sylvanus).

International audienceIn forest-specialist mammals, forest loss may induce resistance to animal movement and reduce gene flow between populations, and thereby increase genetic erosion and extinction risks for populations. Understanding how landscape features affect gene flow is of critical importance for conservation. Using landscape genetic tools at multiple spatial scales, we assessed the effects of landscape heterogeneity (in particular the presence of wide open or rural habitats) on gene flow in an endangered forest-specialist species – the Barbary macaque (Macaca sylvanus) –, in its major forest site in Morocco. We genotyped 248 individuals from 23 macaque groups using 11 microsatellite loci. We modelled different scenarios of isolation by landscape resistance. We further tested the relationships between genetic distance and isolation by resistance, after controlling for the effect of isolation by distance. Our results revealed a significant genetic structure and a disruption of gene flow even in geographic proximity. Whatever the spatial scale, remoteness from the forest edge beyond 1km acted as a barrier to macaque movements. In addition, at a fine scale, human-dominated areas were also detected as a barrier. The detection of private alleles in each population suggests an ongoing process of isolation. The preservation of the Barbary macaque implies 1) strictly avoiding all silvicultural practices (in particular clear-cutting of holm oak forests) that could contribute to increase distances between forest patches, 2) restoring corridors between forests, 3) and preserving key small forest patches as potential stepping stones facilitating macaque dispersal

Paraffin-embedded tissue is less accurate than frozen section analysis for determining VHL mutational status in sporadic renal cell carcinoma.

International audienceINTRODUCTION: Literature controversies exist regarding the prognostic value of VHL mutations. The objective was to compare paraffin-embedded and frozen section specimens for VHL mutations detection and to evaluate the reliability of DNA analysis in formalin-fixed tissues. METHODS: Seventy-six patients with clear cell renal cell carcinoma (RCC) previously assessed for VHL status from frozen samples were included. Seventy-three tumor samples were known to be mutated for VHL. DNA was extracted and an electrophoresis was performed to determine DNA quality. The whole coding sequence was synthesized by double PCR amplification followed by sequencing. Sequencing results were compared with those previously determined from frozen samples. RESULTS: DNA could be extracted from the 76 paraffin samples. DNA quality was highly degraded and significantly less amplified by PCR in 34.2%, resulting in no sequence available for analysis in 57.7% and discordance with frozen samples in 42.3% of the cases respectively. VHL mutations were found in 52.1% of the whole paraffin samples whereas 98% were mutated; 72% could be sequenced, resulting in 69.1% of VHL mutations in this subset. Only half of observed mutations were fully consistent with frozen analysis in the 3 exons. Neomutations were found in 10.5% and 28.9% of known mutations in frozen samples were not detected in paraffin blocks. Only DNA quality significantly influenced PCR amplification and sequencing. CONCLUSION: Tumoral DNA extraction and VHL mutation analysis can be performed from formalin-fixed paraffin-embedded (FFPE) tissue in RCC. But mutations identified tissues are not strictly concordant with those from frozen analysis and therefore results obtained from FFPE samples should be interpreted with care