Linking membrane physical properties and low temperature tolerance in arthropods

Maintenance of membrane fluidity is of crucial importance in ectotherms experiencing thermal changes. This maintenance has in ectotherms most often been indicated using indirect measures of biochemical changes of phospholipid membranes, which is then assumed to modulate the physico-chemical properties of the membrane. Here, we measure bending rigidity characterizing the membrane flexibility of re-constituted membrane vesicles to provide a more direct link between membrane physical characteristics and low temperature tolerance. Bending rigidity of lipid bilayers was measured in vitro using Giant Unilamellar Vesicles formed from phospholipid extracts of the springtail, Folsomia candida. The bending rigidity of these membranes decreased when exposed to 0.4 vol% ethanol (0.23 mM/L). Springtails exposed to ethanol for 24 h significantly increased their cold shock tolerance. Thus, by chemically inducing decreased membrane rigidity, we have shown a direct link between the physico-chemical properties of the membranes and the capacity to tolerate low temperature in a chill-susceptible arthropod.     

Regeneration in the tail fan of crayfish

Summary The tail fan of a crayfish consists of the caudal end of the body, the telson, and the most caudal limbs, the uropods. We investigated the positional information in these structures with grafting operations. The uropods are biramous; they bifurcate to a lateral exopodite and a medial endopodite. After the distal part of a uropod ramus was grafted to the stump of a ramus, medio-lateral or dorso-ventral mismatch of surfaces provoked the production of supernumerary distal parts. Proximo-distal intercalation between exopodite and endopodite yielded a mosaic ramus. The results show that the two rami contain equivalent ramus fields in congruent orientation. The exopodite consists of basal and distal segments; each of these segments seems to have an equivalent segmental field.The telson regenerated an ablated distal portion poorly, unlike the limbs of crayfish. After the posterior lobe of the telson was inverted dorso-ventrally and grafted into the telson stump, supernumerary posterior lobes regenerated dorsal and ventral to the graft. Thus the dorsal and ventral surfaces of the telson embody different positional information. A grafted uropod endopodite or exopodite healed to the telson, but dorsoventral inversion of the graft did not provoke the formation of supernumerary structures at the graft-host boundary. Because supernumeraries did not form, the relations between positional information in the telson (a body axis structure) and the uropod (a limb) remain unclear.     

Immunocytochemical,electrophoresis, and immunoblot analysis of Heliothis virescens gap junctions isolated in the presence and absence of protease inhibitors

Gap junction-enriched fractions were prepared from larvae of the tobacco budworm Heliothis virescens using the NaOH procedure in the presence or absence of protease inhibitors and were analyzed by SDS-PAGE, immunoblotting and EM immunocytochemistry. Protease inhibitor fractions contained a 48-kDa protein in addition to the 10 proteins in fractions with and without inhibitors. Three polyclonal antibodies were used as probes for gap junction plaques and proteins: R16, against an 40-kDa candidate gap junction protein from Drosophila melanogaster; R17, against the 40-kDa candidate gap junction protein from H. virescens; and R18AP, an affinity purified antibody against a consensus sequence of N-terminal amino acids 2–21 of the H. virescens 40-kDa protein. R16, R17, and R18AP stain the 40- and 48-kDa proteins, R16 and R18AP stain a 64-kDa protein, and R16 stains an 30-kDa protein in the absence of inhibitors. Inclusion of protease inhibitors had no effect on gap junction ultrastructure. R16 and R17 label gap junction plaques in crude membrane and NaOH fractions, whereas R18AP exhibits only a low level of reactivity with gap junctions in crude membrane fractions and none with gap junctions in NaOH fractions. The results show that the 30-, 40-, 48- and 64-kDa proteins are immunologically related and are associated with gap junctions in H. virescens, the N-terminus of the 40-kDa protein is relatively inaccessible or easily lost, and the 48-kDa protein is protease-sensitive.     

Identification of Nitric Oxide-Sensitive and -Insensitive Forms of Cytoplasmic Guanylate Cyclase

Abstract: Cytoplasmic, nitric oxide-activated guanylate cyclases are expressed in many regions of the mammalian brain and are thought to participate in functions as diverse as synaptogenesis and long-term potentiation. In this study, we have characterized cytoplasmic guanylate cyclases in the nervous system of an invertebrate, the American lobster. Cytoplasmic cyclase specific activity is higher in lobster nerve cord than in any other lobster tissue tested, and considerably higher than in typical rat tissues (cerebellum, lung, and liver). However, nitric oxide donors have minimal effects on lobster nerve cord cyclic GMP production, when applied either to intact tissue or to cytoplasmic extracts. Parallel immunocytochemical studies, using an anti-cyclic GMP antibody, reveal that only a small subset of lobster neurons responds to nitric oxide with a significant elevation of cyclic GMP levels. HPLC analysis of nerve cord cytoplasm reveals two chromatographically separable cyclases, a minor nitric oxide-sensitive form whose retention time is identical to that of the conventional mammalian enzyme and a more abundant nitric oxide-insensitive form that appears to be novel. The physiological function and phylogenetic distribution of this nitric oxide-insensitive enzyme, and the signaling mechanisms that regulate its activity, are not known.     

A trophic cascade in a diverse arthropod community caused by a generalist arthropod predator

We tested the hypothesis that a generalist arthropod predator, Tenodera sinensis Saussure, could trigger a trophic cascade in an old-field ecosystem. These mantids had relatively weak effects on abundance and biomass of other carnivorous arthropods as a group. The effect of mantids on herbivores was stronger than on carnivores, mainly concentrated in Homoptera and Diptera. Herbivore load was reduced by mantids with the consequence that overall plant biomass (mainly grasses) was increased. Percapita interaction strengths between mantids and other arthropod taxa were, for the most part, weakly negative. Our study demonstrates that a significant trophic cascade can be triggered by a generalist predator even within the framework of a diverse community with relatively diffuse interactions. Received: 13 January 1997 / Accepted: 28 July 1997     

Early axonogenesis in the embryo of a primitive insect,the silverfish Ctenolepisma longicaudata

The pattern of axon growth from the population of neurons that pioneers the major axon pathways in the central nervous system is highly conserved in winged insects. This study sought to determine whether the same pattern of axon growth is shared by an apterygotic insect, the silverfish. We have found that homologues to at least nine early differentiating winged insect neurons are present in the silverfish. The axon trajectories and the sequence of axon outgrowth from these neurons are very similar in silverfish and winged insects, suggesting that the pterygotic and apterygotic insects share a common developmental Bauplan for the construction of the central nervous system. Some of these neurons do show differences in several aspects of axon growth, including the relative timing of axonogenesis, the polarity of axon growth and the pattern of axon fasciculation. In addition, a major, early-appearing fascicle in the posterior commissure of the silverfish is pioneered by a neuron which does not appear to have an equivalent in the winged insects. These differences are similar in character to, albeit more pronounced than, differences previously reported between two winged insects, the fruitfly Drosophila and the grasshopper. Some of the features of early central axon growth, that set the silverfish embryo apart from the winged insects, are shared by crustacean embryos, providing support for the claim that insects and crustaceans share a common developmental Bauplan for the construction of central axonal pathways.     

Testing the effectiveness of surrogates for assessing biological diversity of arthropods in cereal agricultural landscapes

Agricultural intensification is altering biodiversity patterns worldwide. Rapid and effective methods are needed to monitor these changes in farmland biodiversity, but it becomes both a cost- and time-prohibitive task, particularly for hyper-diverse groups such as arthropods. We evaluated the effectiveness of surrogates in irrigated and rainfed wheat fields in a Mediterranean farmland in NW Spain in order to get a rapid tool to assess arthropod biodiversity. We studied six groups with different ecological needs (i.e. Aphididae, Aphidiinae, Coccinellidae, Formicidae, Heteroptera and Syrphidae) at species level (147 species), genus (105), family (10, only Heteroptera) and order (19) level. Higher taxa, cross-taxa and subset-taxa or total richness approaches were tested as well as the correlation in composition between levels for the selected groups, and the influence of farming regime. Genus richness was a good surrogate of species richness in all six groups studied (R² = 0.38–0.60), like family and order were for Heteroptera (R² = 0.37 and 0.29, respectively). Cross-taxa analyses showed that Aphididae and Aphidiinae genera (R² = 0.19 and 0.30, respectively) and species (R² = 0.20 and 0.28, respectively) were good surrogates for Aphidiinae and Aphididae species respectively. Coccinellidae genera (R² = 0.26) and species (R² = 0.25) were good surrogates for Heteroptera species. Finally, Aphididae and Coccinellidae both at genera (R² = 0.14 and 0.20, respectively) and at species levels (R² = 0.12–0.22, respectively) were good surrogates for total species richness of all groups. Genera composition was the best surrogate for the species composition within each group. Farming regime had no influence on the relationships between surrogates and species patterns in most cases. Our results suggest that genera level is a useful surrogate for all the studied groups and family is appropriate for Heteroptera. Genus level provided a saving of 15% of identification time in Aphididae and 80% for Coccinellidae. This proves its usefulness to asses and monitor biodiversity in wheat croplands and the possibility to reduce costs.     

Arthropod communities in a Japanese cedar (Cryptomeria japonica D. Don) plantation: Abundance,biomass and some properties

Arboreal arthropod communities were censused by insecticidal knockdown in a plantation of Japanese cedar,Cryptomeria japonica, in central Japan from April 1983 to February 1985 at intervals of two months. All arthropods sampled were allocated to higher taxonomic groups or guilds and the composition of the communities was analyzed with respect to abundance, biomass and their seasonal trends. The total densities fluctuated seasonally from 200 to 3500 m⁻²; corresponding biomass values ranged from 7 to 600 mg fresh weight m⁻². Maximum density and biomass generally occurred in summer and were minimum in winter. The consistently dominant guild in terms of abundance were the detritivores, mostly comprising Collembola and oribatid mites, accounting for 36–93% of all arthropods; phytophages, predators and detritivores were major guilds in terms of biomass, showing average proportions of 27%, 23% and 20%, respectively. Biomass ratios of predators to prey were generally high in some canopy communities, suggesting the importance of predation pressure in regulating the population levels of arthropods in forests. Average individual size of predators was approximately proportional to that of prey, irrespective of tree locality. Soil arthropod communities maintained densities approximately 10²–10³ times as large as the corresponding canopy communities throughout the year. The seasonal variations in abundance were much greater in canopy than in soil communities.     

Microfiber reinforcement of an arthropod cuticle

Summary 1. According to its fine structure arthropod cuticle is a laminated composite material. The exoskeleton of a spider is surveyed in an electron microscopical study as to the characteristics of its fiber reinforcement.2. Five types of fiber arrangement are distinguished, ranging from unidirectional to continuously rotating fiber orientation and with three types combining elements of both of these extreme cases. The unidirectional type is rare and its occurrence confined to the innermost part of the walking leg endocuticle and to parts of articular membranes. Quite contrary a continuous and regular rotation of fiber direction is common. In hard cuticle (exocuticle) this type of reinforcement is found exclusively.3. Lamellar width, indicating a 180° rotation of the fiber direction, varies within wide limits. Variation is due to varying numbers of fiber layers and the amount of directional change between them (ca. 0.6–24°, type E).4. The diameter of the chitin/protein microfiber amounts to ca. 35 Å. Its length measured up to 1,5 m in the preparations. A periodicity of ca. 80 Å is noted. The precision of fiber alignment within the cuticle is high.5. In the cuticle of the opisthosoma and the articular membranes endocuticular lamellae are taking a wavy course, which is interpreted to be in relation to the cuticle's softness.6. The mechanical implications of both an increasing number of fiber directions in a composite material and of the different types of fiber reinforcement found in the spider cuticle are demonstrated by a calculation of the directional properties of the moduli of elasticity in tension and shear. The predictable strong influence of both the fiber content and the binder material's E-modulus on the mechanical properties of cuticle is illustrated.7. Instead of a trajectorial design a continuous rotation of the fiber orientation is found in all exocuticles. Thus the spider does not make use of the special advantage (high strength/weight ratio) of unidirectional fiber arrangement in places where it seemed most likely.Many thanks are due to Ms. H. Tscharntke for expert assistance with electron microscopy and photographic work. Dipl.-Ing. F. Och and Dr. G. Haberl (Messerschmitt-Bölkow-Blohm GmbH, Ottobrunn) provided the calculations for Figs. 14 and 15. Their generous help is gratefully acknowledged. Dr. K. Frahm (Universität München, Fakultät f. Tiermedizin) kindly shared his densitometer. Ms. F. Althaus carefully carried out the drawings.