Adipokinetic hormone counteracts oxidative stress elicited in insects by hydrogen peroxide: in vivo and in vitro study

The role of adipokinetic hormone (AKH) in counteracting oxidative stress elicited in the insect body is studied in response to exogenously applied hydrogen peroxide, an important metabolite of oxidative processes. In vivo experiments reveal that the injection of hydrogen peroxide (8 µmol) into the haemocoel of the firebug, Pyrrhocoris apterus L. (Heteroptera: Pyrrhocoridae) increases the level of AKH by 2.8‐fold in the central nervous system (CNS) and by 3.8‐fold in the haemolymph. The injection of hydrogen peroxide also increases the mortality of experimental insects, whereas co‐injection of hydrogen peroxide with Pyrap‐AKH (40 pmol) reduces mortality to almost control levels. Importantly, an increase in haemolymph protein carbonyl levels (i.e. an oxidative stress biomarker) elicited by hydrogen peroxide is decreased by 3.6‐fold to control levels when hydrogen peroxide is co‐injected with Pyrap‐AKH. Similar results are obtained using in vitro experiments. Oxidative stress biomarkers such as malondialdehyde and protein carbonyls are significantly enhanced upon exposure of the isolated CNS to hydrogen peroxide in vitro, whereas co‐treatment of the CNS with hydrogen peroxide and Pyrap‐AKH reduces levels significantly. Moreover, a marked decrease in catalase activity compared with controls is recorded when the CNS is incubated with hydrogen peroxide. Incubation of the CNS with hydrogen peroxide and Pyrap‐AKH together curbs the negative effect on catalase activity. Taken together, the results of the present study provide strong support for the recently published data on the feedback regulation between oxidative stressors and AKH action, and implicate AKH in counteracting oxidative stress. The in vitro experiments should facilitate research on the mode of action of AKH in relation to oxidative stress, and could help clarify the key pathways involved in this process.     

Déroulement des gamétogenèses chez les Crabes Carcinus maenas (L.) et C. mediterraneus Czerniavsky parasités par la Sacculine

A histological study of the gonads and androgenic glands of Carcinus parasitized with Sacculina carcini showed no correlation between the degree of infestation of the gonads by the roots of the parasite and the progressive inhibition of the course of male and female gametogeneses. Also, no correlation could be observed between the degree of degeneration of the androgenic glands and that of the testes.

Modifications in parasitized crab testes essentially involve a degeneration of the mesoderm tissue of the germinative zone; primary gonia become pycnotic after an abnormal development; germ cells already engaged in spermatogenesis become blocked in prophase of meiosis; and ultimately, the testis becomes empty. Progressive degeneration of the androgenic glands, after an initial hypertrophy that starts as early as the internal Sacculina stage, seems related to the disorganization of the neurosecretory centers of the host.

Arrest of spermatogenesis at a more or less advanced stage is probably the result of an imbalance in neurohormonal controls acting directly on the germinative zone or indirectly via the androgenic glands. At the level of the ovaries, parasitic infestation leads to inhibition of the second phase of vitellogenesis possibly due to an abnormality in the organization of the vitellogenic follicles. Hormonal imbalances related to the presence of the parasite appear to be responsible for these phenomena.     

Alterations in light-induced stomatal opening in a starch-deficient mutant of Arabidopsis thaliana L. deficient in chloroplast phosphoglucomutase activity

Stomatal responses to light of Arabidopsis thaliana wild-type plants and mutant plants deficient in starch (phosphoglucomutase deficient) were compared in gas exchange experiments. Stomatal density, size and ultrastructure were identical for the two phenotypes, but no starch was observed in guard cells of the mutant plants whatever the time of day. The overall extent of changes in stomatal conductance during 14 h light–10 h dark cycles was similar for the two phenotypes. However, the slow endogenous stomatal opening occurring in darkness in the wild type was not observed in the mutant plants. Stomata in the mutant plants responded much more slowly to blue light (70 μmol m^?2 s^?1) though the response to red light (250 μmol m^?2 s^?1) was similar to that of wild-type plants. In paradermal sections, stomatal responses to red light (300 μmol m^?2 s^?1) were weak for wild-type plants as well as for mutant plants. Stomatal opening was greater under low blue light (75 μmol m^?2 s^?1) than under red light for the two genotypes. However, in mutant plants, a high chloride concentration (50 mol m^?3) was necessary to achieve the same stomatal aperture as observed for the wild-type plants. These results suggest that starch metabolism, via the synthesis of a counter-ion to potassium (probably malate), is required for full stomatal response to blue light but is not involved in the stomatal response to red light.     

Ecological differentiation between the Sardinian endemic Maniola nurag and the pan-European M. jurtina

Recently refined evolutionary theories have highlighted that ecological interactions and environmental gradients can play a major role in speciation. This paper reports on a 3‐year field study, in which the ecology of two congeneric butterfly species was used to explore and compare the environmental factors determining their spatial distribution. These data are discussed in the context of possible speciation scenarios between the Sardinian populations of Maniola nurag and M. jurtina. M. nurag is endemic to the island of Sardinia, while M. jurtina is widespread over Europe. In Sardinia, the two species are locally sympatric. Mark–release–recapture experiments were combined with measures of environmental variables in 15 1‐ha plots, established in areas of potential habitat for the butterflies. Constrained linear models were parameterized from mark–recapture data to estimate both individual (survival and capture probabilities) and population (population size and recruitment) parameters. The two species had similar demography, movement patterns, life history, and behaviour. Population sizes developed in a parabolic fashion from beginning to end of the flight season. Differences included local population size, adult phenology, and habitat requirements. Long‐distance movements larger than 1.5 km were observed, suggesting a substantial amount of gene‐flow between populations of the endemic as well as the widespread species. Multivariate analyses revealed four main environmental gradients responsible for the abundance of the butterflies in an area. Both species responded similarly to environmental variables. However, each species’s abundance was correlated with a different environmental gradient determined by vegetation cover and structure. When sympatric, the two species responded to subtle differences in microhabitat structure. This might originally have induced their divergence. This study is an example of how empirical field data on population dynamics, dispersal, and habitat characteristics of two sympatric congeners can further our understanding of how species differentiate despite existing gene‐flow. © 2006 The Linnean Society of London, Biological Journal of the Linnean Society, 2006, 89 , 561–574.     

Turnover and accessibility of a reentrant loop of the Na+-glutamate transporter GltS are modulated by the central cytoplasmic loop

GltS of Escherichia coli is a secondary transporter that catalyzes Na+-glutamate symport. The structural model of GltS shows two homologous domains with inverted membrane topology that are connected by a central loop that resides in the cytoplasm. Each domain contains a reentrant loop structure. Accessibility of the Cys residues in two GltS mutants in which Pro351 and Asn356 in the reentrant loop in the C-terminal domain were replaced by Cys is demonstrated to be sensitive to the catalytic state supporting a role for the reentrant loops in catalysis. Saturating concentrations of the substrate L-glutamate protected both mutants against inactivation by thiol reagents, while the presence of the co-ion Na+ stimulated the inactivation of both mutants. Insertion of the 10 kDa biotin acceptor domain (BAD) of oxaloacetate decarboxylase of Klebsiella pneumoniae in the central cytoplasmic loop blocked the access pathway from the periplasmic side of the membrane to the cysteine residues in mutants P351C and N356C in the reentrant loop. Kinetically, insertion of BAD increased the maximal rate of uptake 2.7-fold while leaving the apparent affinity constants for L-glutamate and Na+ unaltered. The data suggests that insertion of BAD in the central loop results in conformational changes at the translocation site that lower the activation energy of the translocation step without affecting the access pathway from the periplasmic side for substrate and co-ions. It is concluded that changes in the central loop that connects the two domains may have a regulatory function on the activity of the transporter.     

SO2-dependent cation competition and compartmentalization in Norway spruce needles

Ion contents in needles from Norway spruce trees [Picea abies (L.) Karst.] growing in Würzburg and in the SO₂-polluted Erzgebirge mountains were analysed to quantify cations which accumulate together with sulphate. In Würzburg there was a positive correlation of potassium (0.680 ± 0.300 Eq Eq^?1 SO₄^?2), magnesium (0.415 ± 0.111 Eq Eq^?1 SO₄^?2) and zinc (0.059 ± 0.006 Eq Eq^?1 SO₄^2?). In the Erzgebirge, potassium was also the stoichiometrically most important cation (0–887 ± 0–180 Eq K⁺ Eq^?1 SO₄^2?). All other correlations examined were weak or statistically non-significant. At both sites the calcium content of spruce needles did not depend on the sulphate content. The lack of a role for Ca²⁺ in neutralizing sulphate is a consequence of the presence of free oxalic acid in needles. Soluble oxalic acid precipitates Ca²⁺, which thereby becomes unavailable as a counterion for SO₄^2?. The activity coefficients of Ca²⁺ and oxalate^2?, and the solubility product of Ca-oxalate, were determined from in vivo data. It is concluded that the chronic accumulation of atmospheric sulphate in spruce needle vacuoles depletes available potassium and thereby strongly interferes with spruce growth and canopy turnover. This leads to impaired spruce vitality, even at sites where acute SO₂ disease symptoms are absent.     

Elevated CO2 enhances stomatal responses to osmotic stress and abscisic acid in Arabidopsis thaliana

A , carbon assimilation rate
ABA, abscisic acid
Ci , intercellular space CO₂ concentration
g , leaf conductance
WUE, water use efficiency

Carbon dioxide and abscisic acid (ABA) are two major signals triggering stomatal closure. Their putative interaction in stomatal regulation was investigated in well-watered air-grown or double CO₂-grown Arabidopsis thaliana plants, using gas exchange and epidermal strip experiments. With plants grown in normal air, a doubling of the CO₂ concentration resulted in a rapid and transient drop in leaf conductance followed by recovery to the pre-treatment level after about two photoperiods. Despite the fact that plants placed in air or in double CO₂ for 2 d exhibited similar levels of leaf conductance, their stomatal responses to an osmotic stress (0·16–0·24 MPa) were different. The decrease in leaf conductance in response to the osmotic stress was strongly enhanced at elevated CO₂. Similarly, the drop in leaf conductance triggered by 1 μ M ABA applied at the root level was stronger at double CO₂. Identical experiments were performed with plants fully grown at double CO₂. Levels of leaf conductance and carbon assimilation rate measured at double CO₂ were similar for air-grown and elevated CO₂-grown plants. An enhanced response to ABA was still observed at high CO₂ in pre-conditioned plants. It is concluded that: (i) in the absence of stress, elevated CO₂ slightly affects leaf conductance in A. thaliana ; (ii) there is a strong interaction in stomatal responses to CO₂ and ABA which is not modified by growth at elevated CO₂.