Complex effects of environment and Wolbachia infections on the life history of Drosophila melanogaster hosts

Wolbachia bacteria are common endosymbionts of many arthropods found in gonads and various somatic tissues. They manipulate host reproduction to enhance their transmission and confer complex effects on fitness-related traits. Some of these effects can serve to increase the survival and transmission efficiency of Wolbachia in the host population. The Wolbachia–Drosophila melanogaster system represents a powerful model to study the evolutionary dynamics of host–microbe interactions and infections. Over the past decades, there has been a replacement of the ancestral wMelCS Wolbachia variant by the more recent wMel variant in worldwide D. melanogaster populations, but the reasons remain unknown. To investigate how environmental change and genetic variation of the symbiont affect host developmental and adult life-history traits, we compared effects of both Wolbachia variants and uninfected controls in wild-caught D. melanogaster strains at three developmental temperatures. While Wolbachia did not influence any developmental life-history traits, we found that both lifespan and fecundity of host females were increased without apparent fitness trade-offs. Interestingly, wMelCS-infected flies were more fecund than uninfected and wMel-infected flies. By contrast, males infected with wMel died sooner, indicating sex-specific effects of infection that are specific to the Wolbachia variant. Our study uncovered complex temperature-specific effects of Wolbachia infections, which suggests that symbiont–host interactions in nature are strongly dependent on the genotypes of both partners and the thermal environment

A simple and effective method for ultrastructural analysis of mitosis in Drosophila S2 cells

© 2016 The AuthorsThe Drosophila S2 tissue culture cells are a widely used system for studies on mitosis. S2 cells are particularly sensitive to gene silencing by RNA interference (RNAi), allowing targeted inactivation of mitotic genes. S2 cells are also well suited for high-resolution light microscopy analysis of mitosis in fixed cells, and can be easily immunostained to detect mitotic components. In addition, S2 cells are amenable to transformation with plasmid encoding fluorescently tagged mitotic proteins, allowing in vivo analysis of their behavior throughout cell division. However, S2 cells have not been widely used for transmission electron microscopy (TEM) analysis, which provides ultrastructural details on the morphology of the mitotic apparatus that cannot be obtained with high-resolution confocal microscopy. Here, we describe a simple method for the ultrastructural analysis of mitosis in Drosophila S2 cells. • Our method, which involves fixation and sectioning of a cell pellet, provides excellent preservation of mitotic structures and allows analysis of a higher number of mitotic divisions per sample, compared to correlative light-electron microscopy.• Dividing cells are randomly oriented within the pellet and are sectioned along different planes, providing all-around information on the structure of the mitotic apparatus

Separation of Flip and Non-Flip parst of Charge Exchange np->pn at energies Tn = 0.5 - 2.0 GeV

The new Delta-Sigma experimental data on the ratio $R_{dp}$ allowed separating the Flip and Non-Flip parts of the differential cross section of $np\to pn$ charge exchange process at the zero angle by the Dean formula. The PSA solutions for the $np\to np$ elastic scattering are transformed to the $np\to pn$ charge exchange representation using unitary transition, and good agreement is obtain.Comment: 7 pages, 2 figure

Long Period Variables in the Large Magellanic Cloud from the EROS-2 survey

Context. The EROS-2 survey has produced a database of millions of time series from stars monitored for more than six years, allowing to classify some of their sources into different variable star types. Among these, Long Period Variables (LPVs), known to follow sequences in the period-luminosity diagram, include long secondary period variables whose variability origin is still a matter of debate. Aims.We use the 856 864 variable stars available from the Large Magellanic Cloud (LMC) in the EROS-2 database to detect, classify and characterize LPVs. Methods. Our method to extract LPVs is based on the statistical Abbe test. It investigates the regularity of the light curve with respect to the survey duration in order to extract candidates with long-term variability. The period search is done by Deeming, Lomb-Scargle and generalized Lomb-Scargle methods, combined with Fourier series fit. Color-magnitude, period-magnitude and period-amplitude diagrams are used to characterize our candidates. Results. We present a catalog of 43 551 LPV candidates for the Large Magellanic Cloud. For each of them, we provide up to five periods, mean magnitude in EROS-2, 2MASS and Spitzer bands, BE-RE color, RE amplitude and spectral type.We use infrared data to make the distinction between RGB, O-rich, C-rich and extreme AGB stars. Properties of our LPV candidates are investigated by analyzing period-luminosity and period-amplitude diagrams.Comment: Accepted for publication in A&

Wolbachia has subtle effects on thermal preference in highly inbred Drosophila melanogaster which vary with life stage and environmental conditions

Abstract Temperature fluctuations are challenging for ectotherms which are not able to regulate body temperature by physiological means and thus have to adjust their thermal environment via behavior. However, little is yet known about whether microbial symbionts influence thermal preference (T p) in ectotherms by modulating their physiology. Several recent studies have demonstrated substantial effects of Wolbachia infections on host T p in different Drosophila species. These data indicate that the direction and strength of thermal preference variation is strongly dependent on host and symbiont genotypes and highly variable among studies. By employing highly controlled experiments, we investigated the impact of several environmental factors including humidity, food quality, light exposure, and experimental setup that may influence T p measurements in adult Drosophila melanogaster flies. Additionally, we assessed the effects of Wolbachia infection on T p of Drosophila at different developmental stages, which has not been done before. We find only subtle effects of Wolbachia on host T p which are strongly affected by experimental variation in adult, but not during juvenile life stages. Our in-depth analyses show that environmental variation has a substantial influence on T p which demonstrates the necessity of careful experimental design and cautious interpretations of T p measurements together with a thorough description of the methods and equipment used to conduct behavioral studies

Reliance of Wolbachia

Wolbachia are gram-negative, obligate, intracellular bacteria carried by a majority of insect species worldwide. Here we use a Wolbachia-infected Drosophila cell line and genome-wide RNA interference (RNAi) screening to identify host factors that influence Wolbachia titer. By screening an RNAi library targeting 15,699 transcribed host genes, we identified 36 candidate genes that dramatically reduced Wolbachia titer and 41 that increased Wolbachia titer. Host gene knockdowns that reduced Wolbachia titer spanned a broad array of biological pathways including genes that influenced mitochondrial function and lipid metabolism. In addition, knockdown of seven genes in the host ubiquitin and proteolysis pathways significantly reduced Wolbachia titer. To test the in vivo relevance of these results, we found that drug and mutant inhibition of proteolysis reduced levels of Wolbachia in the Drosophila oocyte. The presence of Wolbachia in either cell lines or oocytes dramatically alters the distribution and abundance of ubiquitinated proteins. Functional studies revealed that maintenance of Wolbachia titer relies on an intact host Endoplasmic Reticulum (ER)-associated protein degradation pathway (ERAD). Accordingly, electron microscopy studies demonstrated that Wolbachia is intimately associated with the host ER and dramatically alters the morphology of this organelle. Given Wolbachia lack essential amino acid biosynthetic pathways, the reliance of Wolbachia on high rates of host proteolysis via ubiquitination and the ERAD pathways may be a key mechanism for provisioning Wolbachia with amino acids. In addition, the reliance of Wolbachia on the ERAD pathway and disruption of ER morphology suggests a previously unsuspected mechanism for Wolbachia’s potent ability to prevent RNA virus replication