Thesaurus-based disambiguation of gene symbols

Background  

Massive text mining of the biological literature holds great promise of relating disparate information and discovering new knowledge. However, disambiguation of gene symbols is a major bottleneck.     

Functional studies of signaling pathways in peri-implantation development of the mouse embryo by RNAi

Background

Studies of gene function in the mouse have relied mainly on gene targeting via homologous recombination. However, this approach is difficult to apply in specific windows of time, and to simultaneously knock-down multiple genes. Here we report an efficient method for dsRNA-mediated gene silencing in late cleavage-stage mouse embryos that permits examination of phenotypes at post-implantation stages.

Results

We show that introduction of Bmp4 dsRNA into intact blastocysts by electroporation recapitulates the genetic Bmp4 null phenotype at gastrulation. It also reveals a novel role for Bmp4 in the regulation the anterior visceral endoderm specific gene expression and its positioning. We also show that RNAi can be used to simultaneously target several genes. When applied to the three murine isoforms of Dishevelled, it leads to earlier defects than previously observed in double knock-outs. These include severe delays in post-implantation development and defects in the anterior midline and neural folds at headfold stages.

Conclusion

Our results indicate that the BMP4 signalling pathway contributes to the development of the anterior visceral endoderm, and reveal an early functional redundancy between the products of the murine Dishevelled genes. The proposed approach constitutes a powerful tool to screen the functions of genes that govern the development of the mouse embryo.     

Development of the Canadian beef reference herd for gene mapping studies

A project to map quantitative trait loci (QTL), in beef cattle using a full-sib design was initiated using six Bos taurus breeds. Embryo transfer was used in a large scale, short timeframe experiment to develop this herd for gene mapping. Full-sib families allowed for genetic information to be followed through both the sire and the dam and for both parents to be slaughtered so that carcass quality data could also be obtained from both of them at close to typical slaughter ages. Repeatability of response to superovulation was significant among the 3 flushes per female. Response to superovulation was negatively correlated with backfat of the donor. Crossbred embryos were found to have higher survival than purebred embryos.     

Behavioural and physiological consequences of acute social defeat in growing gilts: effects of the social environment

Endocrine, behavioural and immunologic processes, together with body growth, were evaluated in gilts that were defeated at 10 weeks of age in resident-intruder tests. Immediately after defeat, gilts were either separated from or reunited with a familiar conspecific (litter-mate; always a barrow). Gilts were assigned to one of four treatments: (a) DI: defeat, followed by isolation (separation from original litter-mate; n=8); (b) I: no defeat, isolation (control group; n=9); (c) DP; defeat, followed by pair-housing (reunion with original litter-mate; n=8); and (d) P: no defeat, pair-housing (control group; n=8). The following general conclusions were derived: (1) social defeat caused pronounced short-term elevations in hypothalamic-pituitary-adrenal (HPA) and sympathetic-adrenal medullary activities, and of prolactin levels. Moreover, as soon as 1h after defeat, percentages of blood lymphocytes and neutrophilic granulocytes were, respectively, decreased and increased; (2) social defeat had some long-lasting influence on behaviour and physiology, but isolation predominantly determined responses in the longer term. Defeat, as well as isolation, resulted in increased cardiovascular activities compared to P controls, as observed in a novel object test (NOT: +7 days) and an aversion test (AVT: +14 days). Moreover, defeated as well as isolated gilts did not habituate to a repeated novel environment test (NET: -7, +2 and +7 days) in terms of frequencies of vocalising, whereas P controls did. Isolation, through the separation from any other pig, was responsible for the other observed long-term characteristics, which developed progressively. Isolated gilts showed high mobilities and high cortisol responses in the repeated NET (+7 days), not being habituated. This contrasted the reactions of pair-housed gilts, which were much reduced. In addition to their high cardiovascular activities in the NOT and the AVT, isolated gilts also displayed higher heart rates in the repeated NET and during human presence following the NOT, compared to pair-housed gilts. Finally, isolated gilts were more inhibited to approach a novel object (in the NOT) than pair-housed pigs; and (3) stress responses of defeated gilts were modulated by the subsequent social environment. Stimulation of the HPA-axis (plasma- and salivary cortisol) was prolonged in those defeated gilts which were isolated (observed in the first hour). Changes in leucocyte subsets were still observed after 3 days in DI, but were 'normalised' within 1 day in DP gilts. Two days after defeat, habituation to the repeated NET in terms of mobility and salivary cortisol responses occurred in control and DP gilts, but not in DI gilts. We argue that these effects of the social environment shortly after defeat were related to a stress-reducing effect of a stable social relationship, i.e. social support.     

Variability in vocal and behavioural responses to visual isolation between full-sibling families of beef calves

In order to assess the value of vocal response as an indicator of welfare in cattle, it is necessary to investigate factors which influence vocal behaviour of individuals, that are independent of the specific environmental conditions to which they are exposed. The effect of parental differences, sex, age and weight on vocal responses of calves to visual isolation were examined. The relationship between vocal response and the amount of body movement during isolation were also evaluated.

Seventeen full-sibling families of beef calves (N=130) were created by breeding five sires with 13 superovulated dams. Embryos were transferred into unrelated cows, which reared the calves to weaning age. Vocal response of calves and body movement were measured on four occasions, while they were visually isolated for 1 min on a single animal scale platform. The first two observations were made on the day of weaning (mean age 166 days) and the following day. At the third and fourth observations their average ages were 278 and 350 days, respectively. On the four occasions 33.3, 34.8, 27.8 and 38.6% of calves, respectively, vocalized. Both sire and family had a significant influence on the number of vocalizations and acoustic properties of the sounds. Electronically-measured movement while on the scale was also influenced by sire and family, but there was no relationship between vocal responses and body movement. There was a tendency (P=0.08) for older and therefore heavier calves to produce longer vocalizations. In the final observing session, age and weight were positively correlated with fundamental frequency and the sound pressure level at the frequency of greatest intensity, and negatively correlated with the loudest harmonic. At the last two observations, significantly more heifer calves than bull calves vocalized. There was little difference between the acoustic properties of heifers’ and bulls’ vocalizations.

The considerable individual variability in vocal response within a group of identically-treated calves is partly due to parental genetic influences that are independent of rearing and early experience. Other endogenous characteristics such as sex, age and weight also influence vocal response. But there is no clear relationship between individual temperament (as assessed by the amount of body movement during isolation) and characteristics of vocal response.     

Surfactant Bilayers Maintain Transmembrane Protein Activity

In vitro studies of membrane proteins are of interest only if their structure and function are significantly preserved. One approach is to insert them into the lipid bilayers of highly viscous cubic phases rendering the insertion and manipulation of proteins difficult. Less viscous lipid sponge phases are sometimes used, but their relatively narrow domain of existence can be easily disrupted by protein insertion. We present here a sponge phase consisting of nonionic surfactant bilayers. Its extended domain of existence and its low viscosity allow easy insertion and manipulation of membrane proteins. We show for the first time, to our knowledge, that transmembrane proteins, such as bacteriorhodopsin, sarcoplasmic reticulum Ca²⁺ATPase (SERCA1a), and its associated enzymes, are fully active in a surfactant phase.     

A likelihood‐based approach for assessment of extra‐pair paternity and conspecific brood parasitism in natural populations

Genotypes are frequently used to assess alternative reproductive strategies such as extra‐pair paternity and conspecific brood parasitism in wild populations. However, such analyses are vulnerable to genotyping error or molecular artefacts that can bias results. For example, when using multilocus microsatellite data, a mismatch at a single locus, suggesting the offspring was not directly related to its putative parents, can occur quite commonly even when the offspring is truly related. Some recent studies have advocated an ad‐hoc rule that offspring must differ at more than one locus in order to conclude that they are not directly related. While this reduces the frequency with which true offspring are identified as not directly related young, it also introduces bias in the opposite direction, wherein not directly related young are categorized as true offspring. More importantly, it ignores the additional information on allele frequencies which would reduce overall bias. In this study, we present a novel technique for assessing extra‐pair paternity and conspecific brood parasitism using a likelihood‐based approach in a new version of program cervus . We test the suitability of the technique by applying it to a simulated data set and then present an example to demonstrate its influence on the estimation of alternative reproductive strategies.     

Forecasting fish distribution along stream networks: brown trout (Salmo trutta) in Europe

Aim

Species inhabiting fresh waters are severely affected by climate change and other anthropogenic stressors. Effective management and conservation plans require advances in the accuracy and reliability of species distribution forecasts. Here, we forecast distribution shifts of Salmo trutta based on environmental predictors and examine the effect of using different statistical techniques and varying geographical extents on the performance and extrapolation of the models obtained.

Location

Watercourses of Ebro, Elbe and Danube river basins (c. 1,041,000 km²; Mediterranean and temperate climates, Europe).

Methods

The occurrence of S. trutta and variables of climate, land cover and stream topography were assigned to stream reaches. Data obtained were used to build correlative species distribution models (SDMs) and forecasts for future decades (2020s, 2050s and 2080s) under the A1b emissions scenario, using four statistical techniques (generalised linear models, generalised additive models, random forest, and multivariate adaptive regression).

Results

The SDMs showed an excellent performance. Climate was a better predictor than stream topography, while land cover characteristics were not necessary to improve performance. Forecasts predict the distribution of S. trutta to become increasingly restricted over time. The geographical extent of data had a weak impact on model performance and gain/loss values, but better species response curves were generated using data from all three basins collectively. By 2080, 64% of the stream reaches sampled will be unsuitable habitats for S. trutta, with Elbe basin being the most affected, and virtually no new habitats will be gained in any basin.

Main conclusions

More reliable predictions are obtained when the geographical data used for modelling approximate the environmental range where the species is present. Future research incorporating both correlative and mechanistic approaches may increase robustness and accuracy of predictions.