Spectral Unmixing: Analysis of Performance in the Olfactory Bulb In Vivo

Background: The generation of transgenic mice expressing combinations of fluorescent proteins has greatly aided the reporting of activity and identification of specific neuronal populations. Methods capable of separating multiple overlapping fluorescence emission spectra, deep in the living brain, with high sensitivity and temporal resolution are therefore required. Here, we investigate to what extent spectral unmixing addresses these issues. Methodology/Principal Findings: Using fluorescence resonance energy transfer (FRET)-based reporters, and two-photon laser scanning microscopy with synchronous multichannel detection, we report that spectral unmixing consistently improved FRET signal amplitude, both in vitro and in vivo. Our approach allows us to detect odor-evoked FRET transients 180-250 mm deep in the brain, the first demonstration of in vivo spectral imaging and unmixing of FRET signals at depths greater than a few tens of micrometer. Furthermore, we determine the reporter efficiency threshold for which FRET detection is improved by spectral unmixing. Conclusions/Significance: Our method allows the detection of small spectral variations in depth in the living brain, which is essential for imaging efficiently transgenic animals expressing combination of multiple fluorescent proteins

Divergent selection predating the Last Glacial Maximum mainly acted on macro-phenotypes in Norway spruce

The current distribution and population structure of many species were, to a large extent, shaped by cycles of isolation in glacial refugia and subsequent population expansions. Isolation in and postglacial expansion through heterogeneous environments led to either neutral or adaptive divergence. Norway spruce is no exception, and its current distribution is the consequence of a constant interplay between evolutionary and demographic processes. We investigated population differentiation and adaptation of Norway spruce for juvenile growth, diameter of the stem, wood density, and tracheid traits at breast height. Data from 4461 phenotyped and genotyped Norway spruce from 396 half-sib families in two progeny tests were used to test for divergent selection in the framework of Q(ST) vs. F-ST. We show that the macroscopic resultant trait (stem diameter), unlike its microscopic components (tracheid dimensions) and juvenile growth, was under divergent selection that predated the Last Glacial Maximum. Altogether, the current variation in these phenotypic traits in Norway spruce is better explained by local adaptation to ancestral environments than to current ones, where populations were partly preadapted, mainly through growth-related traits

Teasing apart the joint effect of demography and natural selection in the birth of a contact zone

Vast population movements induced by recurrent climatic cycles have shaped the genetic structure of plant species. During glacial periods species were confined to low-latitude refugia from which they recolonized higher latitudes as the climate improved. This multipronged recolonization led to many lineages that later met and formed large contact zones. We utilize genomic data from 5000 Picea abies trees to test for the presence of natural selection during recolonization and establishment of a contact zone in Scandinavia. Scandinavian P. abies is today made up of a southern genetic cluster originating from the Baltics, and a northern one originating from Northern Russia. The contact zone delineating them closely matches the limit between two major climatic regions. We show that natural selection contributed to its establishment and maintenance. First, an isolation-with-migration model with genome-wide linked selection fits the data better than a purely neutral one. Second, many loci show signatures of selection or are associated with environmental variables. These loci, regrouped in clusters on chromosomes, are often related to phenology. Altogether, our results illustrate how climatic cycles, recolonization and selection can establish strong local adaptation along contact zones and affect the genetic architecture of adaptive traits

Porcine Reproductive and Respiratory Syndrome Virus Type 1.3 Lena Triggers Conventional Dendritic Cells 1 Activation and T Helper 1 Immune Response Without Infecting Dendritic Cells

Porcine Reproductive and Respiratory Syndrome virus (PRRSV) is an arterivirus responsible for highly contagious infection and huge economic losses in pig industry. Two species, PRRSV-1 and PRRSV-2 are distinguished, PRRSV-1 being more prevalent in Europe. PRRSV-1 can further be divided in subtypes. PRRSV-1.3 such as Lena are more pathogenic than PRRSV-1.1 such as Lelystad or Flanders13. PRRSV-1.3 viruses trigger a higher Th1 response than PRRSV-1.1, although the role of the cellular immune response in PRRSV clearance remains ill defined. The pathogenicity as well as the T cell response inductions may be differentially impacted according to the capacity of the virus strain to infect and/or activate DCs. However, the interactions of PRRSV with in vivo-differentiated-DC subtypes such as conventional DC1 (cDC1), cDC2, and monocyte-derived DCs (moDC) have not been thoroughly investigated. Here, DC subpopulations from Lena in vivo infected pigs were analyzed for viral genome detection. This experiment demonstrates that cDC1, cDC2, and moDC are not infected in vivo by Lena. Analysis of DC cytokines production revealed that cDC1 are clearly activated in vivo by Lena. In vitro comparison of 3 Europeans strains revealed no infection of the cDC1 and cDC2 and no or little infection of moDC with Lena, whereas the two PRRSV-1.1 strains infect none of the 3 DC subtypes. In vitro investigation of T helper polarization and cytokines production demonstrate that Lena induces a higher Th1 polarization and IFNγ secretion than FL13 and LV. Altogether, this work suggests an activation of cDC1 by Lena associated with a Th1 immune response polarization

Feline low-grade alimentary lymphoma: an emerging entity and a potential animal model for human disease

Background: Low-grade alimentary lymphoma (LGAL) is characterised by the infiltration of neoplastic T-lymphocytes, typically in the small intestine. The incidence of LGAL has increased over the last ten years and it is now the most frequent digestive neoplasia in cats and comprises 60 to 75% of gastrointestinal lymphoma cases. Given that LGAL shares common clinical, paraclinical and ultrasonographic features with inflammatory bowel diseases, establishing a diagnosis is challenging. A review was designed to summarise current knowledge of the pathogenesis, diagnosis, prognosis and treatment of feline LGAL. Electronic searches of PubMed and Science Direct were carried out without date or language restrictions. Results: A total of 176 peer-reviewed documents were identified and most of which were published in the last twenty years. 130 studies were found from the veterinary literature and 46 from the human medicine literature. Heterogeneity of study designs and outcome measures made meta-analysis inappropriate. The pathophysiology of feline LGAL still needs to be elucidated, not least the putative roles of infectious agents, environmental factors as well as genetic events. The most common therapeutic strategy is combination treatment with prednisolone and chlorambucil, and prolonged remission can often be achieved. Developments in immunohistochemical analysis and clonality testing have improved the confidence of clinicians in obtaining a correct diagnosis between LGAL and IBD. The condition shares similarities with some diseases in humans, especially human indolent T-cell lymphoproliferative disorder of the gastrointestinal tract. Conclusions: The pathophysiology of feline LGAL still needs to be elucidated and prospective studies as well as standardisation of therapeutic strategies are needed. A combination of conventional histopathology and immunohistochemistry remains the current gold-standard test, but clinicians should be cautious about reclassifying cats previously diagnosed with IBD to lymphoma on the basis of clonality testing. Importantly, feline LGAL could be considered to be a potential animal model for indolent digestive T-cell lymphoproliferative disorder, a rare condition in human medicine

Multilocus Approach of the genome in the population genetics models

La génétique des populations est l’étude de l’évolution des fréquences alléliques au sein d’une population et de l’influence des pressions évolutives sur ces fréquences. Au sein de cette discipline, des modèles de population et des mesures génétiques sont développés pour pouvoir expliquer et prédire les données génétiques. Toutefois, au fur et à mesure des avancées technologiques, de nouveaux types de données sont disponibles, et il devient primordial de développer de nouveaux modèles et de nouvelles mesures pour pouvoir expliquer ces nouvelles données génétiques, plus denses et plus riches en marqueurs génétiques grâce à l’avènement de techniques comme la Next Generation Sequencing. Pour ce faire, nous proposons dans cette thèse de développer de nouvelles mesures avec une approche dite multilocus, qui considère le génome comme un tout plutôt que comme un agglomérat de locus indépendants. Dans un premier temps, nous avons tenté de construire une base théorique de l’approche multilocus en génétique des populations. Ensuite, nous avons illustré une telle approche à travers l’étude de l’identité par descendance, des graphes de recombinaison ancestraux et des autocorrélogrammes dans les modèles de génétique des populations. À travers ces différentes études de cas, nous avons tenté d’identifier les principaux enjeux et questions que soulève la génétique des populations multilocus.Population genetics is the study of the evolution of allelic frequencies within a population and the influence of evolutionary pressures on these frequencies. Within this field, one could develop population models and measures to explain and predict genetic data. However, as technologie evolves new types of data are available, and it becomes essential to develop new models and new measures to reflect these new genetic marker data, increasingly richer and denser thanks to the advent of new techniques such as the Next Generation Sequencing. To this end, we propose in this thesis to develop new measures with the so-called multilocus approach, which considers the genome as a whole rather than an agglomerate of independent loci. We have first tried to build a theoretical basis for the multilocus approach in population genetics. Then, we have illustrated this multilocus approach with the case studies of identity by descent, ancestral recombination graphs and autocorrelograms in population genetics models. Through these different studies, we tried to identify the main issues and questions that the multilocus population genetics raises

Approche multilocus du génome dans les modèles de génétique des populations

Population genetics is the study of the evolution of allelic frequencies within a population and the influence of evolutionary pressures on these frequencies. Within this field, one could develop population models and measures to explain and predict genetic data. However, as technologie evolves new types of data are available, and it becomes essential to develop new models and new measures to reflect these new genetic marker data, increasingly richer and denser thanks to the advent of new techniques such as the Next Generation Sequencing. To this end, we propose in this thesis to develop new measures with the so-called multilocus approach, which considers the genome as a whole rather than an agglomerate of independent loci. We have first tried to build a theoretical basis for the multilocus approach in population genetics. Then, we have illustrated this multilocus approach with the case studies of identity by descent, ancestral recombination graphs and autocorrelograms in population genetics models. Through these different studies, we tried to identify the main issues and questions that the multilocus population genetics raises.La génétique des populations est l’étude de l’évolution des fréquences alléliques au sein d’une population et de l’influence des pressions évolutives sur ces fréquences. Au sein de cette discipline, des modèles de population et des mesures génétiques sont développés pour pouvoir expliquer et prédire les données génétiques. Toutefois, au fur et à mesure des avancées technologiques, de nouveaux types de données sont disponibles, et il devient primordial de développer de nouveaux modèles et de nouvelles mesures pour pouvoir expliquer ces nouvelles données génétiques, plus denses et plus riches en marqueurs génétiques grâce à l’avènement de techniques comme la Next Generation Sequencing. Pour ce faire, nous proposons dans cette thèse de développer de nouvelles mesures avec une approche dite multilocus, qui considère le génome comme un tout plutôt que comme un agglomérat de locus indépendants. Dans un premier temps, nous avons tenté de construire une base théorique de l’approche multilocus en génétique des populations. Ensuite, nous avons illustré une telle approche à travers l’étude de l’identité par descendance, des graphes de recombinaison ancestraux et des autocorrélogrammes dans les modèles de génétique des populations. À travers ces différentes études de cas, nous avons tenté d’identifier les principaux enjeux et questions que soulève la génétique des populations multilocus

Multilocus identity by descent in population genetics: models and predictions

<p>Please find the pseudo dataset (in "data.tgz") of the evolution of Identity-by-descent in a Wright-Fisher population model, with drift as the only evolutionary pressure, with a constant population size, and panmictic without selfing. The individuals are diploids and the initial population are unrelated and non inbred, meaning that chromosome in the initial population are pairwisely different.</p> <p>In "data.tgz", one could find stored all the IBD blocks at specific generations (1,000,000 replicates, over 500 generations, population size of 20 diploid individuals). For each file, the first column is the number of the replicate, the second the number of the individual in the replicate and the third the length of the IBD block. Each line is an IBD block.</p> <p>This dataset was generated with the joint program "ibd-static", with the command line "./ibd -i 20 -r 1000000 -g $g -sa", where$g is the number of generations ranging from 1 to 500.</p> <p>The output file could be summarised in several mean values, what is done by "mfile.R", and the resulting file is "xmean".</p

Blocks of chromosomes identical by descent in a population: Models and predictions.

With the highly dense genomic data available nowadays, ignoring linkage between genes would result in a huge loss of information. One way to prevent such a loss is to focus on the blocks of chromosomes shared identical by descent (IBD) in populations. The development of the theoretical framework modelling IBD processes is essential to support the advent of new tools such as haplotype phasing, imputation, inferring population structure and demographic history, mapping loci or detecting signatures of selection. This article aims to present the relevant models used in this context, and specify the underlying definitions of identity by descent that are yet to be gathered at one place. In light of this, we derived a general expression for the expected IBD block length, for any population model at any generation after founding

How to reach a higher selection plateau with optimal contribution selection and compensatory mating?

In breeding programs, balancing short-term genetic gain and loss of diversity per generation is essential to sustain a long-term genetic response. Depending on the dynamic of the species, the acceptable trade-off will be different. One of the most common and successful tools to achieve this management is the Optimal Contribution Selection (OCS), which readily mathematically formulate the trade-off between genetic gain and coancestry. However, OCS only accounts for the next generation gain and diversity, which can lead to suboptimality given the uncertainties of random mating and segregation. In this paper, we have extended the OCS by conveniently integrating a way to promote certain parental pairs, so that this method can account for the next t+2 generation. In the study case of Populus nigra , fully phenotyped and SNP array genotyped, we have shown that (i) a non negligible part of the long-term success of a breeding strategy depends on the implemented mating strategy, and (ii) favoring a compensatory mating can accelerate the selection without compromising the future diversity