We previously identified Nob1 as a quantitative trait locus for high-fat diet-induced obesity and diabetes in genome-wide scans of outcross populations of obese and lean mouse strains. Additional crossbreeding experiments indicated that Nob1 represents an obesity suppressor from the lean Swiss Jim Lambert (SJL) strain. Here we identify a SJL-specific mutation in the Tbc1d1 gene that results in a truncated protein lacking the TBC Rab-GTPase-activating protein domain. TBC1D1, which has been recently linked to human obesity, is related to the insulin signaling protein AS160 and is predominantly expressed in skeletal muscle. Knockdown of TBC1D1 in skeletal muscle cells increased fatty acid uptake and oxidation, whereas overexpression of TBC1D1 had the opposite effect. Recombinant congenic mice lacking TBC1D1 showed reduced body weight, decreased respiratory quotient, increased fatty acid oxidation and reduced glucose uptake in isolated skeletal muscle. Our data strongly suggest that mutation of Tbc1d1 suppresses high-fat diet-induced obesity by increasing lipid use in skeletal muscle.

The predicted increase in frequency and severity of heat waves due to climate change is expected to alter disease dynamics by reducing hosts' ability to resist infections. This could take place via two different mechanisms: (1) through general reduction in hosts' performance under harsh environmental conditions and/or (2) through altered resource allocation that reduces expression of defense traits in order to maintain other traits. We tested these alternative hypotheses by measuring the effect of an experimental heat wave (25 vs. 15°C) on the constitutive level of immune defense (hemocyte concentration, phenoloxidase [PO]-like activity, antibacterial activity of hemolymph), and life history traits (growth and number of oviposited eggs) of the great pond snail Lymnaea stagnalis. We also manipulated the exposure time to high temperature (1, 3, 5, 7, 9, or 11 days). We found that if the exposure to high temperature lasted <1 week, immune function was not affected. However, when the exposure lasted longer than that, the level of snails' immune function (hemocyte concentration and PO-like activity) was reduced. Snails' growth and reproduction increased within the first week of exposure to high temperature. However, longer exposures did not lead to a further increase in cumulative reproductive output. Our results show that short experimental heat waves do not alter immune function but lead to plastic responses that increase snails' growth and reproduction. Thus, although the relative expression of traits changes, short experimental heat waves do not impair snails' defenses. Negative effects on performance get pronounced when the heat waves are prolonged suggesting that high performance cannot be maintained over long time periods. This ultimately reduces the levels of defense traits.

The glucose transporter GLUT8 cycles between intracellular vesicles and the plasma membrane. Like the insulin-responsive glucose transporter GLUT4, GLUT8 is primarily located in intracellular compartments under basal conditions. Whereas translocation of GLUT4 to the plasma membrane is stimulated by insulin, the distribution of GLUT8 is not affected by insulin treatment in adipose cells. However, blocking endocytosis by co-expression of a dominant-negative dynamin GTPase (K44A) or mutation of the N-terminal dileucine (LL12/13) motif in GLUT8 leads to accumulation of the glucose transporter at the cell surface in a variety of different cell types. Yeast two-hybrid analyses and GST pulldown assays reveal that the LL signal constitutes a binding site for the β2-adaptin subunit of the heterotetrameric AP-2 adaptor complex, implicating this motif in targeting of GLUT8 to clathrin-coated vesicles. Moreover, yeast two-hybrid assays provide evidence that the binding site for the LL motif maps to the appendage domain of β2-adaptin. To analyze the biological significance of the LL/β2 interaction, we utilized RNA interference to specifically knockdown AP-2. Our results show that RNAi-mediated targeting of the μ2 subunit leads to cellular depletion of AP-2, but not AP-1 adaptor complexes in HeLa cells. As a consequence, GLUT8 accumulates at the plasma membrane at comparable levels to those observed in K44A-transfected cells. Conversely, the intracellular localization of mutant GLUT8-LL/AA is restored by replacing the LL motif in GLUT8 with the transferrin receptor-derived μ2-adaptin binding motif YTRF, indicating that for endocytosis both AP-2 binding motifs can substitute for each other. Thus, our data demonstrate that recruitment of GLUT8 to the endocytic machinery occurs via direct interaction of the dileucine motif with β2-adaptin, and that endocytosis might be the main site at which GLUT8 is likely to be regulated.

BackgroundOn-going global climate change poses a serious threat for natural populations unless they are able to evolutionarily adapt to changing environmental conditions (e.g. increasing average temperatures, occurrence of extreme weather events). A prerequisite for evolutionary change is within-population heritable genetic variation in traits subject to selection. In relation to climate change, mainly phenological traits as well as heat and desiccation resistance have been examined for such variation. Therefore, it is important to investigate adaptive potential under climate change conditions across a broader range of traits. This is especially true for life-history traits and defences against natural enemies (e.g. parasites) since they influence organisms’ fitness both directly and through species interactions. We examined the adaptive potential of fitness-related traits and their responses to heat waves in a population of a freshwater snail, Lymnaea stagnalis. We estimated family-level variation and covariation in life history (size, reproduction) and constitutive immune defence traits [haemocyte concentration, phenoloxidase (PO)-like activity, antibacterial activity of haemolymph] in snails experimentally exposed to typical (15 °C) and heat wave (25 °C) temperatures. We also assessed variation in the reaction norms of these traits between the treatments.ResultsWe found that at the heat wave temperature, snails were larger and reproduced more, while their immune defence was reduced. Snails showed high family-level variation in all examined traits within both temperature treatments. The only negative genetic correlation (between reproduction and antibacterial activity) appeared at the high temperature. However, we found no family-level variation in the responses of most examined traits to the experimental heat wave (i.e. largely parallel reaction norms between the treatments). Only the reduction of PO-like activity when exposed to the high temperature showed family-level variation, suggesting that the cost of heat waves may be lower for some families and could evolve under selection.ConclusionOur results suggest that there is genetic potential for adaptation within both thermal environments and that trait evolution may not be strongly affected by trade-offs between them. However, rare differences in thermal reaction norms across families indicate limited evolutionary potential in the responses of snails to changing temperatures during extreme weather events.Electronic supplementary materialThe online version of this article (doi:10.1186/s12862-017-0988-x) contains supplementary material, which is available to authorized users.

Lymnaea stagnalis is a holarctic freshwater snail inhabiting shallow littoral zones of stagnant waters such as lakes and ponds. It is an SUMMARY Understanding the outcomes of host-parasite interactions in nature is in high demand as parasites and pathogens are important for several ecological and evolutionary processes. Ecological immunology (ecoimmunology) has a key role in reaching this goal because immune defence is the main physiological barrier against infections. To date, ecoimmunological studies largely lean on measuring constitutive immune defences (components of defence that are always active). However, understanding the role of inducible components of immune function is important as the immune system is largely an inducible defence. Measuring such defences can be complicated as different parasites may activate different immune cascades, and expression of different immune traits may not be independent. We examined the suitability of different immune activation techniques for the freshwater snail Lymnaea stagnalis. By experimentally challenging snails with different immune elicitors [injection with snail saline (i.e. wounding), lyophilized Escherichia coli cells, lyophilized Micrococcus lysodeikticus cells, healthy snail gonad, and trematodeinfected snail gonad; maintenance in microorganism-enriched water] and measuring phenoloxidase-like and antibacterial activity of their haemolymph, we found increased immune activity against some immune elicitors, but also decreased activity. Our findings suggest potentially complicated relationships among immune traits, and propose suitable techniques for ecological studies in this study system.

BackgroundExtreme weather events such as summer heat waves become more frequent owing to global climate change and are predicted to alter disease dynamics. This is because high temperatures can reduce host immune function. Predicting the impact of climate change on host-parasite interactions is, however, difficult as temperature may also affect parasite infective stages and other host characteristics determining the outcome of interaction.MethodsTwo experiments were conducted to investigate these phenomena in a Lymnaea stagnalis–Echinoparyphium aconiatum (Trematoda) interaction. In the first experiment, the effects of exposure of snails to experimental heat waves [maintenance at 25°C vs. 15°C (control)] with different durations (3 days, 7 days) on the infection success of parasite cercariae was examined. In the second experiment, the infection success was examined under similar conditions, while controlling for the possible temperature effects on cercariae and at least partly also for host physiological changes that take place rapidly compared to alterations in immune function (exposure to cercariae at intermediate 20°C).ResultsIn the first experiment, increased infection success at 25°C was found independently of the duration of the heat wave. In the second experiment, increased infection success was found only in snails maintained at 25°C for 7 days, a treatment in which snail immune defence is known to be impaired.ConclusionsThese results suggest that the effects of host resistance in determining overall parasite infection success can be overridden by effects of temperature on parasite transmission stages and/or alterations in other host traits than immune defence.

Jyv€ askyl€ a, Jyv€ askyl€ a, Finland Summary 1. Environmental factors impairing physiological condition of organisms are assumed to predispose them to parasite infections. This is because host immune function is typically condition-dependent. However, poor physiological condition has been reported to reduce host susceptibility to parasites in various systems. 2. We examined whether such an effect can be due to altered exposure of hosts to active parasite transmission stages by investigating chemo-orientation of free-swimming cercariae larvae of a parasite Echinoparyphium aconiatum towards its snail host Lymnaea stagnalis. 3. We used both long-term and short-term feeding treatments to manipulate the body condition and physiological traits related to food processing in experimental (i.e. target) snails and measured the preference of cercariae towards snail-conditioned water (SCW) over clean water. 4. We found that chemo-orientation of cercariae depended on the nutritional status of target snails. High physiological condition (long-term feeding) attracted parasites, but cercariae did not show preference towards SCW from individuals in poor physiological condition (long-term starvation). Food processing (short-term feeding treatments) did not affect chemo-orientation. 5. Our results suggest that host condition-dependent chemo-orientation of parasite larvae is a likely mechanism explaining the reduced susceptibility of snails to infection due to food limitation in our study system. In general, the use of condition-dependent host cues can be highly beneficial for parasites as it increases their transmission to high-quality hosts. Furthermore, evolving counter adaptations to such a transmission strategy can be very difficult for hosts.

Global climate change imposes a serious threat to natural populations of many species. Estimates of the effects of climate change‐mediated environmental stresses are, however, often based only on their direct effects on organisms, and neglect the potential transgenerational (e.g. maternal) effects. We tested whether high temperature (i.e. an experimental heatwave), which is known to reduce the performance of adult Lymnaea stagnalis snails, affects the produced offspring (eggs and hatchlings) through maternal effects, and how strong these effects are compared with the effects of direct exposure of offspring to high temperature. We examined the effect of maternal thermal environment (15°C versus 25°C) on per offspring investment (egg size), and the role of both maternal and offspring thermal environments (15°C versus 25°C) on hatching success and developmental time of eggs, offspring survival after hatching, and hatchling size at the age of 5 weeks. Exposure of mothers to high temperature reduced the size of oviposited eggs, increased their hatching success, and also made the onset of hatching earlier. However, high maternal temperature reduced the survival and the final size of hatched juveniles. Direct exposure of offspring to high temperature reduced their survival (both eggs and hatchlings) but increased the developmental rate and growth of those individuals that survived. Interestingly, the magnitude of maternal effects on hatching success of eggs and hatchling survival were similar to the direct effects of high temperature. Our results indicate that heatwaves can affect natural populations through transgenerational maternal effects and that the magnitude of those effects can be equally strong to the direct effects of temperature, although this depends on the trait considered. These findings highlight the importance of considering the transgenerational effects of climate warming when estimating its effects in the wild.