Comparison of NUCLEOCOUNTER, ANDROVISION with Leja chambers and the newly developed ANDROVISION eFlow for sperm concentration analysis in boars.

Exact analysis of sperm concentration in raw and diluted semen is of major importance in swine artificial insemination, as sperm concentration is one of the most important characteristics of an ejaculate determining the value of the ejaculate and the productive life of the boar. The study compares different methods for sperm concentration analysis in raw and diluted boar semen: NUCLEOCOUNTER SP-100, the ANDROVISION with Leja chambers and the new ANDROVISION eFlow system. The Concordance Correlation Coefficient (CCC) between NUCLEOCOUNTER and ANDROVISION eFlow was 0.955 for raw (n = 185 ejaculates) and 0.94 for diluted semen (n = 109 ejaculates). The CCC between NUCLEOCOUNTER and ANDROVISION with Leja chambers was 0.66. A Bland-Altman plot of split-sample measurements of sperm concentration with NUCLEOCOUNTER and ANDROVISION eFlow showed that 95.1% of all measurements lay within ± 1.96 standard deviation. The coefficients of variance were 1.6 ± 1.3%, 3.6 ± 3.6% and 7.3 ± 6.3% for NUCLEOCOUNTER, ANDROVISION eFlow and ANDROVISION with Leja chambers in diluted semen, respectively. NUCLEOCOUNTER and ANDROVISION eFlow are comparable tools to measure the concentration of raw and diluted boar semen. In comparison to ANDROVISION with Leja chambers, concentration analyses of diluted semen using NUCLEOCOUNTER or ANDROVISION eFlow show a higher repeatability within and a higher concordance between the methods.

Lipid content influences division of labour in a clonal ant.

The fat body, a major metabolic hub in insects, is involved in many functions, e.g. energy storage, nutrient sensing and immune response. In social insects, fat appears to play an additional role in division of labour between egg layers and workers, which specialize in non-reproductive tasks inside and outside their nest. For instance, reproductives are more resistant to starvation, and changes in fat content have been associated with the transition from inside to outside work or reproductive activities. However, most studies have been correlative and we still need to unravel the causal interrelationships between fat content and division of both reproductive and non-reproductive labour. Clonal ants, e.g. Platythyrea punctata, are ideal models for studying task partitioning without confounding variation in genotype and morphology. In this study, we examined the range of variation and flexibility of fat content throughout the lifespan of workers, the threshold of corpulence associated with foraging or reproduction and whether low fat content is a cause rather than a consequence of the transition to foraging. We found that lipid stores change with division of labour from corpulent to lean and, in reverted nurses, back to corpulent. In addition, our data show the presence of fat content thresholds that trigger the onset of foraging or egg-laying behaviour. Our study supports the view that mechanisms that regulate reproduction and foraging in solitary insects, in particular the nutritional status of individuals, have been co-opted to regulate division of labour in colonies of social insects.

Dose dependent gene expression is dynamically modulated by the history, physiology and age of yeast cells.

Cells respond to external stimuli with transient gene expression changes in order to adapt to environmental alterations. However, the dose response profile of gene induction upon a given stress depends on many intrinsic and extrinsic factors. Here we show that the accurate quantification of dose dependent gene expression by live cell luciferase reporters reveals fundamental insights into stress signaling. We make the following discoveries applying this non-invasive reporter technology. (1) Signal transduction sensitivities can be compared and we apply this here to salt, oxidative and xenobiotic stress responsive transcription factors. (2) Stress signaling depends on where and how the damage is generated within the cell. Specifically we show that two ROS-generating agents, menadione and hydrogen peroxide, differ in their dependence on mitochondrial respiration. (3) Stress signaling is conditioned by the cells history. We demonstrate here that positive memory or an acquired resistance towards oxidative stress is induced dependent on the nature of the previous stress experience. (4) The metabolic state of the cell impinges on the sensitivity of stress signaling. This is shown here for the shift towards higher stress doses of the response profile for yeast cells moved from complex to synthetic medium. (5) The age of the cell conditions its transcriptional response capacity, which is demonstrated by the changes of the dose response to oxidative stress during both replicative and chronological aging. We conclude that capturing dose dependent gene expression in real time will be of invaluable help to understand stress signaling and its dynamic modulation.

Stress and early experience underlie dominance status and division of labour in a clonal insect.

Cooperation and division of labour are fundamental in the 'major transitions' in evolution. While the factors regulating cell differentiation in multi-cellular organisms are quite well understood, we are just beginning to unveil the mechanisms underlying individual specialization in cooperative groups of animals. Clonal ants allow the study of which factors influence task allocation without confounding variation in genotype and morphology. Here, we subjected larvae and freshly hatched workers of the clonal ant Platythyrea punctata to different rearing conditions and investigated how these manipulations affected division of labour among pairs of oppositely treated, same-aged clonemates. High rearing temperature, physical stress, injury and malnutrition increased the propensity of individuals to become subordinate foragers rather than dominant reproductives. This is reflected in changed gene regulation: early stages of division of labour were associated with different expression of genes involved in nutrient signalling pathways, metabolism and the phenotypic response to environmental stimuli. Many of these genes appear to be capable of responding to a broad range of stressors. They might link environmental stimuli to behavioural and phenotypic changes and could therefore be more broadly involved in caste differentiation in social insects. Our experiments also shed light on the causes of behavioural variation among genetically identical individuals.