A Genome-Wide Association Study Points out the Causal Implication of SOX9 in the Sex-Reversal Phenotype in XX Pigs

Among farm animals, pigs are known to show XX sex-reversal. In such cases the individuals are genetically female but exhibit a hermaphroditism, or a male phenotype. While the frequency of this congenital disease is quite low (less than 1%), the economic losses are significant for pig breeders. These losses result from sterility, urogenital infections and the carcasses being downgraded because of the risk of boar taint. It has been clearly demonstrated that the SRY gene is not involved in most cases of sex-reversal in pigs, and that autosomal recessive mutations remain to be discovered. A whole-genome scan analysis was performed in the French Large-White population to identify candidate genes: 38 families comprising the two non-affected parents and 1 to 11 sex-reversed full-sib piglets were genotyped with the PorcineSNP60 BeadChip. A Transmission Disequilibrium Test revealed a highly significant candidate region on SSC12 (most significant p-value<4.65.10^-10) containing the SOX9 gene. SOX9, one of the master genes involved in testis differentiation, was sequenced together with one of its main regulatory region Tesco. However, no causal mutations could be identified in either of the two sequenced regions. Further haplotype analyses did not identify a shared homozygous segment between the affected pigs, suggesting either a lack of power due to the SNP properties of the chip, or a second causative locus. Together with information from humans and mice, this study in pigs adds to the field of knowledge, which will lead to characterization of novel molecular mechanisms regulating sexual differentiation and dysregulation in cases of sex reversal.     

The Protein Kinase TPL2 Is Essential for ERK1/ERK2 Activation and Cytokine Gene Expression in Airway Epithelial Cells Exposed to Pathogen-Associated Molecular Patterns (PAMPs)

The epithelium forms a physical barrier important to the detection of pathogens. P. aeruginosa infections are frequently encountered in Cystic Fibrosis lungs, lead to ERK1/ERK2 activation and contribute to tissue destruction. We report here that in bronchial airway epithelial cells (BEAS-2B), diffusible material from P. aeruginosa and TLR2, TLR3 and TLR5 ligands activates ERK1/ERK2 via the protein kinase TPL2 and not the growth factor receptor EGFR. Activation of TPL2 by these agonists in airway epithelial cells requires the protein kinases TAK1 and IKKβ in accordance with the previously reported model of activation of TPL2 in macrophages. Inhibition of TPL2 activity with a pharmacological inhibitor (Compound 1) not only prevented ERK1/ERK2 activation but also decreased cytokine synthesis in response to pathogen-associated molecular patterns. These results suggest that inhibition of the protein kinase TPL2 is an attractive strategy to decrease inflammation in the lungs when it is not warranted.     

Population expansion and breeding success of Bearded Vultures Gypaetus barbatus in the French Pyrenees: results from long‐term population monitoring

Based on monitoring of Bearded Vultures over 24 years in the French Pyrenees, we assessed factors explaining temporal and spatial variations in numbers and breeding performance. The number of territorial pairs increased throughout the study period from 16 in 1994 to 44 in 2017. No significant negative trends in mean productivity (fledglings per territorial pair) were detected with increasing population size. Colonization probability increased significantly with breeding population size the previous year and with the regular provision of supplementary food in the territory the winter when colonization occurred. Colonization of new territories simultaneously increased the distribution range and local densities, but we found no effect of number of near neighbours on productivity. Pairs having bred less than 5 years together had a much lower probability of laying clutches, and higher lay rates were observed inside or close to protected areas after accounting for pair‐bond length, so productivity of territories inside protected areas was significantly higher. Nest success decreased with advanced lay date and increased with winter food abundance. Nesting failures in the study area were frequently associated with harsh weather. Additionally, disturbance by human activities was the second most important identified cause of breeding failure. The probability of failing due to disturbance was higher in western areas (where breeding areas are more accessible to humans), outside protected areas, and has increased with time. After a failure due to disturbance, there was a significantly higher probability of not producing a clutch the following year as compared with pairs that had not failed or had failed due to other causes, indicating deferred effects of disturbance. Our results show the benefits of conservation management actions, such as implementation of protected areas or designed supplementary food programmes in winter, to help range expansion. On the other hand, we did not find a significant effect of winter supplementary food on productivity. Management of feeding sites should be adapted to more specific planning, being used only in areas where natural food availability is scarce, avoiding its use close to breeding sites when juveniles disperse, and targeted mainly to help range expansion. Our results also highlight the importance of maintaining or enhancing good populations of wild ungulates and regulating human activities around nesting sites of this threatened species.     

TPM analyses reveal that FtsK contributes both to the assembly and the activation of the XerCD-dif recombination synapse

Circular chromosomes can form dimers during replication and failure to resolve those into monomers prevents chromosome segregation, which leads to cell death. Dimer resolution is catalysed by a highly conserved site-specific recombination system, called XerCD-dif in Escherichia coli. Recombination is activated by the DNA translocase FtsK, which is associated with the division septum, and is thought to contribute to the assembly of the XerCD-dif synapse. In our study, direct observation of the assembly of the XerCD-dif synapse, which had previously eluded other methods, was made possible by the use of Tethered Particle Motion, a single molecule approach. We show that XerC, XerD and two dif sites suffice for the assembly of XerCD-dif synapses in absence of FtsK, but lead to inactive XerCD-dif synapses. We also show that the presence of the γ domain of FtsK increases the rate of synapse formation and convert them into active synapses where recombination occurs. Our results represent the first direct observation of the formation of the XerCD-dif recombination synapse and its activation by FtsK.     

Identification and distribution of a GABA receptor mutation conferring dieldrin resistance in the malaria vector Anopheles funestus in Africa

Growing problems of pyrethroid resistance in Anopheles funestus have intensified efforts to identify alternative insecticides. Many agrochemicals target the GABA receptors, but cross-resistance from dieldrin resistance may preclude their introduction.Dieldrin resistance was detected in An. funestus populations from West (Burkina Faso) and central (Cameroon) Africa, but populations from East (Uganda) and Southern Africa (Mozambique and Malawi) were fully susceptible to this insecticide. Partial sequencing of the dieldrin target site, the ??-aminobutyric acid (GABA) receptor, identified two amino acid substitutions, A296S and V327I. The A296S mutation has been associated with dieldrin resistance in other species. The V327I mutations was detected in the resistant sample from Burkina Faso and Cameroon and consistently associated with the A296S substitution. The full-length of the An. funestus GABA-receptor gene, amplified by RT-PCR, generated a sequence of 1674 bp encoding 557 amino acid of the protein in An. funestus with 98% similarity to that of Anopheles gambiae. Two diagnostic assays were developed to genotype the A296S mutation (pyrosequencing and PCR-RFLP), and use of these assays revealed high frequency of the resistant allele in Burkina Faso (60%) and Cameroon (82%), moderate level in Benin (16%) while low frequency or absence of the mutation was observed respectively in Uganda (7.5%) or 0% in Malawi and Mozambique.The distribution of the Rdl^R mutation in An. funestus populations in Africa suggests extensive barriers to gene flow between populations from different regions.     

Tasting novel foods and selecting nutrient content in a highly successful ecological invader,the common myna

Invasion success is dependent on the ability of a species to discover and exploit novel food resources. Within this context, individuals must be willing to taste novel foods. They must also be capable of evaluating the nutritional content of new foods, and selecting their relative intake in order to fulfil their nutritional needs. Whereas the former capacity is well studied, little is known about the latter capacity. First, using the common myna as a model avian invader species, we quantified the willingness of mynas to taste novel foods relative to familiar ones. Mynas readily tasted high protein (HP) novel foods and consumed them in higher quantities compared to a familiar food. Data showed that at three different levels – mixes, ingredients and macronutrients – intake could not be explained by a random model. In experiment 2, we confirmed that mynas were making their selection based on protein (P) content rather than a selection for novelty per se. When given the choice of three equally unfamiliar foods, mynas again ate disproportionately from the high protein relative to high lipid and high carbohydrate foods. Analysis revealed that mynas consumed amounts of protein that were closer to the ones in their natural diet. Finally, in experiment 3, we measured inter‐individual variation in innovation and exploration propensities, and examined associations with inter‐individual variation in consumption of specific macronutrients. This analysis revealed that individuals that selected HP pellets were more exploratory and individuals that selected HC pellets were quicker to solve the innovative foraging task. These findings indicate that not only the willingness to taste novel foods, but also the capacity to evaluate their nutritional content, might be central to the myna's substantial ecological success.     

Influence of Media Components and pH on Somatic Embryo Induction in Three Genotypes of Soybean

The influence of media components on the initiation of somatic embryogenesis in three genotypes of soybean was investigated. The following genotypes were used: Iroquois, Macon, and Savoy. Media modifications included sucrose concentration, type and concentration of auxin at two pH levels, and pH level independently. Immature cotyledons were used as the source of explant. Cotyledons were placed on a medium containing MS salts, B5 vitamins, sucrose, and auxin. Gelrite (0.2%) was used as the solidifying agent. Sucrose concentrations of 1, 2, 3, 4.5, or 6% were used. The auxins used included 2,4-dichlorophenoxyacetic acid (2,4-D) and naphthaleneacetic acid (NAA) with each at concentrations of 45.2, 90.4, 135.7, 180.9, and 226.2 M. The pH of each the media was adjusted to either 5.7 or 7.0 with 1 N NaOH. In an additional experiment, the effect of the two pH levels, 5.7 and 7.0, was investigated independently. Overall, the frequency of somatic embryogenesis significantly varied among the different genotypes used in this study, with Iroquois showing the highest response. Frequency of somatic embryogenesis also varied in response to the different treatments used, including sucrose and auxin. The highest initiation (91.7%) and mean number of somatic embryos per responding explant (14.9) of Iroquois was observed in a medium containing 2% sucrose. The highest initiation (97.1%) and mean number of somatic embryos per responding explant (19.5) was observed in Iroquois on 135.7 M 2,4-D and Savoy on 135.7 M 2,4-D, respectively, for the auxin by pH level experiment. No significant differences were observed among the two pH treatments used.     

The mechanism of γ-Secretase dysfunction in familial Alzheimer disease

The mechanisms by which mutations in the presenilins (PSEN) or the amyloid precursor protein (APP) genes cause familial Alzheimer disease (FAD) are controversial. FAD mutations increase the release of amyloid β (Aβ)42 relative to Aβ40 by an unknown, possibly gain‐of‐toxic‐function, mechanism. However, many PSEN mutations paradoxically impair γ‐secretase and ‘loss‐of‐function’ mechanisms have also been postulated. Here, we use kinetic studies to demonstrate that FAD mutations affect Aβ generation via three different mechanisms, resulting in qualitative changes in the Aβ profiles, which are not limited to Aβ42. Loss of ε‐cleavage function is not generally observed among FAD mutants. On the other hand, γ‐secretase inhibitors used in the clinic appear to block the initial ε‐cleavage step, but unexpectedly affect more selectively Notch than APP processing, while modulators act as activators of the carboxypeptidase‐like (γ) activity. Overall, we provide a coherent explanation for the effect of different FAD mutations, demonstrating the importance of qualitative rather than quantitative changes in the Aβ products, and suggest fundamental improvements for current drug development efforts.