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International audience; Soil erosion in freshwaters induces important changes in lake metabolism. The organic matter and the nutrients supplied by soil inputs can change internal biogeochemical cycles and subsidize the whole food web from basal organisms to top-predators. Since the last two decades, the role of allochthonous organic matter as a basal resource for aquatic food webs in natural and controlled conditions has received a growing attention. We studied the impact of soil on the functioning of pond ecosystems by performing monthly additions of soil in freshwater mesocosms. In addition, the food-web structure was manipulated by addition of omnivorous fish to study interactions between the bottom-up effect of soil addition and the top-down effect of fish. The effects of soil and fish addition on the, the elemental and the biochemical compositions of pelagic compartments and recent sediment, on the biomass of seston and zooplankton and on the sediment rates were studied. Soil inputs had no effect on biomass, stoichiometry and lipid composition of seston and zooplankton but fish growth was enhanced by soil addition. Soil treatment had several (but idiosyncratic) effects on the stoichiometry and on the lipid composition of recent sediment. However, the sedimentation rates and the potential biodegradability of recent sediment were not affected by soil inputs. Fish addition affected chlorophyll-a concentration of the water column, seston biomass, sedimentation rates and stoichiometry of seston, zooplankton and short-term sediment. The lipid composition of recent sediment was also influenced by fish addition. However, fish addition did not change the biodegradability of recent sediment. Finally, we did not observe any significant interaction between soil and fish treatments. Our results suggest that the addition of soil as allochthonous inputs to aquatic ecosystem induced a subsidize of the food web only on fish, probably due to direct foraging on bottom sediment. Nevertheless, this process does not induce an effect on the biodegradability of recent sediments. Therefore, allochthonous inputs appear to have little effects on nutrient cycling. The predominance of the effects of food-web structure over allochthonous inputs and the absence of interaction between these top-down and bottom-up controls suggest that the bottom-up control was minor compared to the top-down one exerted by top-predators.

1. Sediment plays a key role in internal nutrient cycling and eutrophication in lakes. However, studies focusing on the efficiency of the biomanipulation techniques for improving the control of primary producers have rarely examined the effects of changes in food-web structure on the sediment biochemical composition and biodegradability. 2. In a 1-year experiment conducted in large replicated mesocosms, we tested how the absence or presence of a zooplanktivorous fish (roach, Rutilus rutilus) affected the elemental composition and the potential biodegradability of recently deposited sediment in a eutrophic system. The potential biodegradability of these sediments was assessed in laboratory microcosms by measuring the production of CO2 during 44-day incubations. 3. The potential biodegradability of recently deposited sediment from the fish treatment was 60% higher than that from the fishless treatment. This higher biodegradability was corroborated by a higher annual loss of sediment in fish enclosures (36%) than in fishless ones (16%). Annual losses of carbon, nitrogen and organic phosphorous were higher for sediment from fish enclosures. 4. Carbon and nitrogen contents of sediment were higher for the fish treatment. In contrast, the sediment C/N ratio, one of the proxies used to estimate sediment biodegradability, did not differ between treatments. No relationship was observed between elemental composition of sediment and its potential biodegradability. This latter appeared to be more probably dependent on the biochemical composition of the sediment and especially on the content of labile compounds such as proteins, sugars and polyunsaturated fatty acids. The use of sterols as biomarkers revealed an important degradation by microorganisms of 1-year-old sediment from both fish and fishless treatments. 5. Our results revealed that fish biomanipulations might favour clear water states not only through a stronger topdown control on phytoplankton but also through a lower biodegradability of sediment reducing internal nutrient cycling.

International audience; 1. Declines in availability of plant resources to pollinators are a major cause of pollinator loss. The management of plant communities to enhance floral resources is often proposed as a way to sustain pollinator populations. Nectar, the main energetic resource for pollinators, plays a central role in behaviour and composition of pollinator communities. Abiotic and biotic factors are known to influence nectar traits at both the species and community levels, but the impact of plant community composition itself has never been investigated. 2. Below-ground interactions in plant communities can induce changes in plant development through (i) plant-derived litter amendment of the soil and (ii) competition for soil resources between plants. We tested how plant below-ground interactions affect above-ground nectar traits involved in plant attractiveness to pollinators. 3. A short-term pot experiment was carried out with three temperate grassland species Mimulus guttatus, Lamium amplexicaule, and Medicago sativa, showing distinct litter stoichiometry and competitive abilities for soil resources. Litter amendment (none, mono and tri-specific litter) and plant interaction treatments (monocultures, two-and three-species mixtures) were crossed in a factorial design. 4. Litter amendment to the soil led to an increase in total nectar sugar content in L. amplexicaule plants but not in the two other species. We also found that the presence of M. guttatus, a competitive species, reduced the total nectar sugar content in L. amplexicaule through a concomitant decrease in nectar volume per flower and in floral display size, but not in other species. Species-specific responses of nectar traits to variation in soil nitrogen availability were thus observed, suggesting consequences for plant species and community attractiveness to pollinators. However, we did not find evidence that the legume M. sativa affected nectar traits of any neighbouring plants. 5. Synthesis. Our results demonstrate that litter inputs and competition between plants for soil resources can alter nectar traits linked to plant attractiveness to pollinators. This supports the idea that below-ground plant-plant interactions for soil resources can influence above-ground plant-plant interactions for pollination services. This offers promising perspectives in studying the role of below-ground-above-ground interactions on higher trophic levels.

In skeletal muscle, sarcoplasmic reticulum (SR) Ca2+ depletion is suspected to trigger a calcium entry across the plasma membrane and recent studies also suggest that the opening of channels spontaneously active at rest and possibly involved in Duchenne dystrophy may be regulated by SR Ca2+ depletion. Here we simultaneously used the cell-attached- and whole-cell- voltage clamp techniques as well as intracellular Ca2+ measurements on single isolated mouse skeletal muscle fibres to unravel any possible change in membrane conductance that would depend upon SR Ca2+ release and/or SR Ca2+ depletion. Delayed rectifier K+ single channel activity was routinely detected during whole-cell depolarizing pulses. In addition the activity of channels carrying unitary inward currents of ~1.5 pA at -80 mV was detected in 17 out of 127 and in 21 out of 59 patches in control and mdx dystrophic fibres, respectively. In both populations of fibres, large whole-cell depolarizing pulses did not reproducibly increase this channel activity. This was also true when, on the long run, repeated application of the whole-cell pulses led to exhaustion of the Ca2+ transient. SR Ca2+ depletion produced by the SR Ca2+ pump inhibitor cyclopiazonic acid (CPA) also failed to induce any increase in the resting whole-cell conductance and in the inward single channel activity. Overall results indicate that voltage-activated SR Ca2+ release and/or SR Ca2+ depletion are not sufficient to activate the opening of channels carrying inward currents at negative voltages and challenge the physiological relevance of a store-operated membrane conductance in adult skeletal muscle

In skeletal muscle, nitric oxide (NO) is commonly referred to as a modulator of the activity of the ryanodine receptor (RyR) calcium release channel. However the reported effects of NO on isolated SR preparations and single RyR activity are diverse, and how NO affects SR calcium release and intracellular calcium homeostasis under physiological conditions remains poorly documented and hardly predictable. Here, we studied the effects of NO donors on membrane current and intracellular [Ca2+] in single skeletal muscle fibres from mouse, under voltage-clamp conditions. When fibres were chronically exposed to millimolar levels of sodium nitroprusside (SNP) and challenged by short membrane depolarizations, there was a progressive increase in the resting [Ca2+] level. This effect was use-dependent with the slope of rise in resting [Ca2+] being increased twice when the depolarizing pulse level was raised from -20 to +10 mV. Analysis of the decay of the [Ca2+] transients suggested that cytoplasmic Ca2+ removal processes were largely unaffected by the presence of SNP. Also the functional properties of the dihydropyridine receptor were very similar under control conditions and in the presence of SNP. The resting [Ca2+] elevation due to SNP was accompanied by a depression of the peak calcium release elicited by pulses to +10 mV. The effects of SNP could be reproduced by the chemically distinct NO donor NOC-12. They could be reversed upon exposure of the fibres to the thiol reducing agent Dithiothreitol. Results suggest that large levels of NO produce a redox-sensitive continuous leak of Ca2+ from the SR, through a limited number of release channels that do not close once they are activated by membrane depolarization. This SR Ca2+ leak and the resulting increase in resting [Ca2+] may be important in mediating the effects of excess NO on voltage activated calcium release.

About 30 genes are predicted to encode degenerin/epithelial sodium channels (DEG/ENaCs) in Caenorhabditis elegans but the gating mode of these channels has not been determined. Using the whole-cell configuration of the patch-clamp technique in acutely dissected C. elegans, we investigated the effects of H+ as a potential activating factor of DEG/ENaCs on electrical properties of body wall muscle cells. Under current-clamp conditions, decreasing external pH from 7.2 to 6.1 led to a reversible depolarization of muscle cells associated with a decrease in input resistance which was partially inhibited by amiloride. Under voltage-clamp conditions, extracellular acidification activated an inward desensitizing current at -60 mV. In the absence of external Ca2+, H+ -gated channels were found to be slightly more permeable to Na+ than to K+ and were blocked by amiloride with a K0.5 of 31 microm at -60 mV. An inward current could be also activated by protons in a GABA receptor null mutant in the presence of D-tubocurare and in an unc-105 null mutant. These results demonstrate that ion channels sharing common properties with mammalian acid-sensing ion channels (ASICs) are functional in C. elegans muscle which should prove useful for understanding proton sensing in animals.

Degenerins have emerged from genetic studies in Caenorhabditis elegans as candidate mechanically gated amiloride-sensitive ion channels for transducing mechanical stimuli into cellular responses. In C. elegans muscle, the existence of a genetic interaction between the unc-105 degenerin gene and let-2, a gene encoding an α2(IV) collagen, raised the possibility that UNC-105 may function as a mechanically gated channel in a stretch receptor complex. However, to date, ion channel activity of UNC-105 has only been recorded in a gain-of-function mutant form in heterologous expression systems. In this study we investigated the in situ properties of UNC-105 using the whole cell configuration of the patch clamp technique on body wall muscle cells from acutely dissected C. elegans. Amiloride was found to be without effect on membrane potential of wild-type muscle cells, suggesting that the UNC-105 degenerin is electrically silent in resting muscle. Hypo-osmotic shocks induced a reversible depolarization of muscle cells but which was not affected by amiloride. Deformation of the cells by applying tension to the filamentous complex on which muscle cells remained attached or by ejecting external solution under pressure failed to induce any change of membrane potential. In gain-of-function unc-105(n506) mutant cells, an amiloride-sensitive inward Na+ current was found to be constitutively active, leading to maintained muscle depolarization. An associated mutation in the α2(IV) collagen LET-2 led to the closure of the mutant UNC-105(n506) channel while a collagenase treatment of these double mutant cells caused it to re-open, giving evidence for a functional interaction between LET-2 collagen and mutant UNC-105 channel.

International audience