Electroantennogram of the American cockroach: effect of oxygen and an electrical model

1. With a view to clarifying the role of oxygen in the olfactory reception of insects, the intact antenna of the male American cockroach (Periplaneta americana) was studied under normal conditions and during reversible anoxia, external or tracheal, using an electroantennogram (EAG) elicited by a sex pheromone stimulation. As a first step, the anatomical and electrical characteristics of the antenna were investigated. 2. Based on the microanatomical study, a cockroach antenna was characterized by structural parameters reflecting the haemolymph and gas exchange in the antenna. Valves in the wall of a haemolymphatic vessel were discovered. 3. The resting DC background voltage (DCBV) and the EAG were continuously recorded and proved to be more strongly affected by tracheal anoxia than by anoxia from the outside. 4. A minimal electrical model of the antenna based on the Thurm-Kaissling-De Kramer equivalent circuit of a sensillum was shown to be valid to explain the origin of the DCBV and EAG as well as their changes after oxygen cutoff and resumption. 5. Two formal opposing processes, excitation and desensitization, probably related to the receptor mechanisms, have been used to interpret the kinetics of the EAG and the effect of anoxia on EAG parameters. The desensitization is thought to be more sensitive to oxygen lack than the excitation.     

Resource use of specialist butterflies in agricultural landscapes: conservation lessons from the butterfly Phengaris (Maculinea) nausithous

Most of the European grassland butterfly species are dependent on species rich grasslands shaped by low intensity farming. Conservation of these specialist species in agricultural landscapes relies on knowledge of their essential resources and the spatial distribution of these resources. In The Netherlands, the dusky large blue Phengaris (Maculinea) nausithous butterflies were extinct until their reintroduction in 1990. In addition, a spontaneous recolonization of road verges in an agricultural landscape occurred in 2001 in the southern part of The Netherlands. We analyzed the use of the essential resources, both host plants and host ants, of the latter population during the summers of 2003 and 2005. First we tested whether the distribution of the butterflies during several years could be explained by both the presence of host plants as well as host ants, as we expected that the resource that limits the distribution of this species can differ between locations and over time. We found that oviposition site selection was related to the most abundant resource. While in 2003, site selection was best explained by the presence of the host ant Myrmica scabrinodis, in 2005 it was more strongly related to flowerhead availability of the host plant. We secondly compared the net displacement of individuals between the road verge population and the reintroduced population in the Moerputten meadows, since we expected that movement of individuals depends on the structure of their habitat. On the road verges, butterflies moved significantly shorter distances than on meadows, which limits the butterflies in finding their essential resources. Finally we analyzed the availability of the two essential resources in the surroundings of the road verge population. Given the short net displacement distances and the adverse landscape features for long-distance dispersal, this landscape analysis suggests that the Phengaris population at the Posterholt site is trapped on the recently recolonized road verges. These results highlight the importance of assessing the availability of essential resources across different years and locations relative to the movement of the butterflies, and the necessity to careful manage these resources for the conservation of specialist species in agricultural landscapes, such as this butterfly species.     

Mechanism of the down regulation of photosynthesis by blue light in the Cyanobacterium synechocystis sp. PCC 6803

Exposure to blue light has previously been shown to induce the reversible quenching of fluorescence in cyanobacteria, indicative of a photoprotective mechanism responsible for the down regulation of photosynthesis. We have investigated the molecular mechanism behind fluorescence quenching by characterizing changes in excitation energy transfer through the phycobilin pigments of the phycobilisome to chlorophyll with steady-state and time-resolved fluorescence excitation and emission spectroscopy. Quenching was investigated in both a photosystem II-less mutant, and DCMU-poisoned wild-type Synechocystis sp. PCC 6803. The action spectra for blue-light-induced quenching was identical in both cell types and was dominated by a band in the blue region, peaking at 480 nm. Fluorescence quenching and its dark recovery was inhibited by the protein cross-linking agent glutaraldehyde, which could maintain cells in either the quenched or the unquenched state. We found that high phosphate concentrations that inhibit phycobilisome mobility and the regulation of energy transfer by the light-state transition did not affect blue-light-induced fluorescence quenching. Both room temperature and 77 K fluorescence emission spectra revealed that fluorescence quenching was associated with phycobilin emission. Quenching was characterized by a decrease in the emission of allophycocyanin and long wavelength phycobilisome terminal emitters relative to that of phycocyanin. A global analysis of the room-temperature fluorescence decay kinetics revealed that phycocyanin and photosystem I decay components were unaffected by quenching, whereas the decay components originating from allophycocyanin and phycobilisome terminal emitters were altered. Our data support a regulatory mechanism involving a protein conformational change and/or change in protein-protein interaction which quenches excitation energy at the core of the phycobilisome.     

Stimulation of bursting in pre-Bötzinger neurons by Epac through calcium release and modulation of TRPM4 and K-ATP channels

The exchange factor directly activated by cAMP (Epac) can couple cAMP production to the activation of particular membrane and cytoplasmic targets. Using patch-clamp recordings and calcium imaging in organotypic brainstem slices, we examined the role of Epac in pre-B?tzinger complex, an essential part of the respiratory network. The selective agonist 8-(4-chlorophenylthio)-2'-O-methyl-cAMP (8-pCPT) sensitized calcium mobilisation from inositol-1,4,5-trisphosphate-sensitive internal stores that stimulated TRPM4 (transient receptor potential cation channel, subfamily M, Melastatin) channels and potentiated the bursts of action potentials. 8-pCPT actions were abolished after inhibition of phospholipase C with U73122 and depletion of calcium stores with thapsigargin. Caffeine-sensitive release channels were not modulated by 8-pCPT. Epac inhibited ATP-sensitive K(+) channels that also led to the enhancement of bursting by 8-pCPT. Bursting activity, spontaneous calcium transients and activity of TRPM4 and ATP-sensitive K(+) channels were potentiated after brief exposures to bradykinin and incubation with wortmannin produced opposite effects that can be explained by changes in phosphatidylinositol 4,5-bisphosphate levels. 8-pCPT stimulated the respiratory motor output in functionally intact preparations and the effects of bradykinin and wortmannin were identical to those observed in organotypic slices. The data thus indicate a novel pathway of controlling bursting activity in pre-B?tzinger complex neurons through Epac that can involved in reinforcement of the respiratory activity by cAMP.     

Effect of exercise and training on phospholemman phosphorylation in human skeletal muscle

Phospholemman (PLM, FXYD1) is a partner protein and regulator of the Na(+)-K(+)-ATPase (Na(+)-K(+) pump). We explored the impact of acute and short-term training exercise on PLM physiology in human skeletal muscle. A group of moderately trained males (n = 8) performed a 1-h acute bout of exercise by utilizing a one-legged cycling protocol. Muscle biopsies were taken from vastus lateralis at 0 and 63 min (non-exercised leg) and 30 and 60 min (exercised leg). In a group of sedentary males (n = 9), we determined the effect of a 10-day intense aerobic cycle training on Na(+)-K(+)-ATPase subunit expression, PLM phosphorylation, and total PLM expression as well as PLM phosphorylation in response to acute exercise (1 h at ～72% Vo(2peak)). Biopsies were taken at rest, immediately following, and 3 h after an acute exercise bout before and at the conclusion of the 10-day training study. PLM phosphorylation was increased both at Ser(63) and Ser(68) immediately after acute exercise (75%, P < 0.05, and 30%, P < 0.05, respectively). Short-term training had no adaptive effect on PLM phosphorylation at Ser(63) and Ser(68), nor was the total amount of PLM altered posttraining. The protein expressions of α(1)-, α(2)-,and β(1)-subunits of Na(+)-K(+)-ATPase were increased after training (113%, P < 0.05, 49%, P < 0.05, and 27%, P < 0.05, respectively). Whereas an acute bout of exercise increased the phosphorylation of PKCα/βII on Thr(638/641) pre- and posttraining, phosphorylation of PKCζ/λ on Thr(403/410) was increased in response to acute exercise only after the 10-day training. In conclusion, we show that only acute exercise, and not short-term training, increases phosphorylation of PLM on Ser(63) and Ser(68), and data from one-legged cycling indicate that this effect of exercise on PLM phosphorylation is not due to systemic factors. Our results provide evidence that phosphorylation of PLM may play a role in the acute regulation of the Na(+)-K(+)-ATPase response to exercise.     

A comparative analysis of metabarcoding and morphology‐based identification of benthic communities across different regional seas

In a world of declining biodiversity, monitoring is becoming crucial. Molecular methods, such as metabarcoding, have the potential to rapidly expand our knowledge of biodiversity, supporting assessment, management, and conservation. In the marine environment, where hard substrata are more difficult to access than soft bottoms for quantitative ecological studies, Artificial Substrate Units (ASUs) allow for standardized sampling. We deployed ASUs within five regional seas (Baltic Sea, Northeast Atlantic Ocean, Mediterranean Sea, Black Sea, and Red Sea) for 12–26 months to measure the diversity and community composition of macroinvertebrates. We identified invertebrates using a traditional approach based on morphological characters, and by metabarcoding of the mitochondrial cytochrome oxidase I (COI) gene. We compared community composition and diversity metrics obtained using the two methods. Diversity was significantly correlated between data types. Metabarcoding of ASUs allowed for robust comparisons of community composition and diversity, but not all groups were successfully sequenced. All locations were significantly different in taxonomic composition as measured with both kinds of data. We recovered previously known regional biogeographical patterns in both datasets (e.g., low species diversity in the Black and Baltic Seas, affinity between the Bay of Biscay and the Mediterranean). We conclude that the two approaches provide complementary information and that metabarcoding shows great promise for marine monitoring. However, until its pitfalls are addressed, the use of metabarcoding in monitoring of rocky benthic assemblages should be used in addition to classical approaches rather than instead of them.     

ADP regulates movements of mitochondria in neurons

Mitochondria often reside in subcellular regions with high metabolic demands. We examined the mechanisms that can govern the relocation of mitochondria to these sites in respiratory neurons. Mitochondria were visualized using tetramethylrhodamineethylester, and their movements were analyzed by applying single-particle tracking. Intracellular ATP ([ATP](i)) was assessed by imaging the luminescence of luciferase, the fluorescence of the ATP analog TNP-ATP, and by monitoring the activity of K(ATP) channels. Directed movements of mitochondria were accompanied by transient increases in TNP-ATP fluorescence. Application of glutamate and hypoxia reversibly decreased [ATP](i) levels and inhibited the directed transport. Injections of ATP did not rescue the motility of mitochondria after its inhibition by hypoxia. Introduction of ADP suppressed mitochondrial movements and occluded the effects of subsequent hypoxia. Mitochondria decreased their velocity in the proximity of synapses that correlated with local [ATP](i) depletions. Using a model of motor-assisted transport and Monte Carlo simulations, we showed that mitochondrial traffic is more sensitive to increases in [ADP](i) than to [ATP](i) depletions. We propose that consumption of synaptic ATP can produce local increases in [ADP](i) and facilitate the targeting of mitochondria to synapses.