Sex-related differences in reaction norms in the butterfly Lycaena tityrus (Lepidoptera: Lycaenidae)

We investigate sexual differences in reaction norms in directly developing individuals of the copper butterfly Lycaena tityrus predicted from sexual selection theory. As recent studies on butterflies revealed a high degree of adaptive plasticity in growth and development, which may undermine the basic trade‐offs assumed in life‐history theory, we focus on effects of temperature, trying to drive growth rates to their physiological upper limit and thus disclosing otherwise potentially concealed responses. Development time strongly depended on temperature, leading in accordance with a central assumption in life‐history theory to a larger size at low temperatures, and vice versa. At all temperatures larval development time of males was significantly shorter compared to females, as was predicted by protandry theory. This was partially due to an invariably higher growth rate of males. However, sexes responded in different ways to developmental time constraints caused by increasing temperatures. Despite the shorter larval time of males, both sexes achieved similar body sizes at lower temperatures, because males avoided a reduction in weight due to plastic growth. At high temperatures, in contrast, males were forced to make a trade‐off in which they favoured early emergence over large size, leading to a dramatic weight loss. Weight of females, however, remained similar throughout showing no trade‐off. These different reaction norms reflect divergent selective pressures acting on males and females, which can be explained in relation to the reproductive system. The strong selection for early emergence in males is likely to be due to monandry, discrete non‐overlapping generations (as was already predicted by theory), and territoriality, because prior ownership of a territory seems to be a major advantage for successful reproduction. On the other hand, the preference of females for large body size was expected due to the close relationship between this trait and fecundity. Thus, our results highlight the extraordinary importance of the specific reproductive system, which can influence central life‐history traits in manifold ways.     

Physiological costs of growing fast: does accelerated growth reduce pay-off in adult fitness?

Accumulating evidence suggests that, in contrast to earlier assumptions, juvenile growth rates are optimised by means of natural and sexual selection rather than maximised to be as fast as possible. Owing to the generally accepted advantage of growing fast to adulthood, such adaptive variation strongly implies the existence of costs attached to rapid growth. By using four populations of protandrous copper butterflies with known differences in intrinsic growth rates within and across populations, we investigate a potential trade-off between rapid growth and the proportionate weight loss at metamorphosis. While controlling for effects of pupal time and mass, we demonstrate that (1) protandrous males, exhibiting higher growth rates, suffer a higher weight loss than females throughout, that (2) population differences in weight loss generally follow known differences in growth rates, and that (3) males have by 6 higher metabolic rates than females during pupal development. These results support the notion that a higher weight loss during the development to adulthood may comprise a physiological cost of rapid development, with the pay-off of accelerated growth being reduced by a disproportionally smaller adult sizeCo-ordinating editor: Leimer     

The cell-cell adhesion molecule EpCAM interacts directly with the tight junction protein claudin-7

We recently described that in the metastasizing rat pancreatic carcinoma line BSp73ASML the cell-cell adhesion molecule EpCAM, CD44 variant isoforms and the tetraspanins D6.1A and CD9 form a complex that is located in glycolipid-enriched membrane microdomains. This complex contains, in addition, an undefined 20 kDa protein. As such complex formation influenced cell-cell adhesion and apoptosis resistance, it became of interest to identify the 20 kDa polypeptide. This 20 kDa protein, which co-precipitated with EpCAM in BSp73ASML lysates, was identified as the tight junction protein claudin-7. Correspondingly, an association between EpCAM and claudin-7 was noted in rat and human tumors and in non-transformed tissues of the gastrointestinal tract. Co-localization of the two molecules was most pronounced at basolateral membranes, but was also observed in tight junctions. Evidence for direct protein-protein interactions between EpCAM and claudin-7 was obtained by co-immunoprecipitation after treatment of tumor cells with a membrane-permeable chemical cross-linker. The complex, which is located in glycolipid-enriched membrane microdomains, is not disrupted by partial cholesterol depletion, but claudin-7 phosphorylation is restricted to the localization in glycolipid-enriched membrane microdomains. This is the first report on an association between EpCAM and claudins in both non-transformed tissues and metastasizing tumor cell lines.     

Assessing ant assemblages: pitfall trapping versus nest counting (Hymenoptera, Formicidae)

Pitfall trapping and nest counting are the most common census methods for ant assemblages. We examined the concordance between pitfall catches and nest counts on dry grassland. Spearman rank correlations and non-metric multidimensional scaling of the Bray Curtis similarity index revealed moderate concordance between the data collated by the two methods, but overall method-related differences were considerable. The dissimilarity was influenced by the type of land management, but not by trapping period or plot shape. Trapping success depended on nest density, ground vegetation cover and species-specific traits (inhabited stratum, colony size, foraging distance). Even when these factors were taken into account, the convertibility of pitfall trap and nest density values was unsatisfactory: the census method proved to be crucial in designing ant-ecological studies and interpreting literature data. Received 11 November 2005; revised 9 February 2006; accepted 1 March 2006.     

Structural and functional diversities among mu-conotoxins targeting TTX-resistant sodium channels

mu-Conotoxins are peptides that block sodium channels. Molecular cloning was used to identify four novel mu-conotoxins: CnIIIA, CnIIIB, CIIIA, and MIIIA from Conus consors, C. catus and C. magus. A comparison of their sequences with those of previously characterized mu-conotoxins suggested that the new mu-conotoxins were likely to target tetrodotoxin-resistant (TTX-r) sodium channels. The four peptides were chemically synthesized, and their biological activities were characterized. The new conotoxins all blocked, albeit with varying potencies, TTX-r sodium currents in frog dorsal-root-ganglion (DRG) neurons. The more potent of the four new mu-conotoxins, CnIIIA and CIIIA, exhibited a strikingly different selectivity profile in blocking TTX-r versus TTX-sensitive channels, as determined by their ability to block extracellularly recorded action potentials in three preparations from frog: skeletal muscle, cardiac muscle and TTX-treated C-fibers. CnIIIA was highly specific for TTX-r sodium channels, whereas CIIIA was nonselective. Both peptides appeared significantly less potent in blocking TTX-r sodium currents in rat and mouse DRG neurons. When CnIIIA and CIIIA were injected intracranially into mice, both induced seizures, but only CIIIA caused paralysis. This is the most comprehensive characterization to date of the structural and functional diversities of an emerging group of mu-conotoxins targeting TTX-r sodium channels.     

Synthetic muO-conotoxin MrVIB blocks TTX-resistant sodium channel NaV1.8 and has a long-lasting analgesic activity

MuO-conotoxin MrVIB is a blocker of voltage-gated sodium channels, including TTX-sensitive and -resistant subtypes. A comprehensive characterization of this peptide has been hampered by the lack of sufficient synthetic material. Here, we describe the successful chemical synthesis and oxidative folding of MrVIB that has made an investigation of the pharmacological properties and therapeutic potential of the peptide feasible. We show for the first time that synthetic MrVIB blocks rat NaV1.8 sodium channels and has potent and long-lasting local anesthetic effects when tested in two pain assays in rats. Furthermore, MrVIB can block propagation of action potentials in A- and C-fibers in sciatic nerve as well as skeletal muscle in isolated preparations from rat. Our work provides the first example of analgesia produced by a conotoxin that blocks sodium channels. The emerging diversity of antinociceptive mechanisms targeted by different classes of conotoxins is discussed.     

Angiopoietin-2 sensitizes endothelial cells to TNF-alpha and has a crucial role in the induction of inflammation

The angiopoietins Ang-1 and Ang-2 have been identified as ligands of the receptor tyrosine kinase Tie-2 (refs. 1,2). Paracrine Ang-1-mediated activation of Tie-2 acts as a regulator of vessel maturation and vascular quiescence. In turn, the antagonistic ligand Ang-2 acts by an autocrine mechanism and is stored in endothelial Weibel-Palade bodies from where it can be rapidly released upon stimulation. The rapid release of Ang-2 implies functions of the angiopoietin-Tie system beyond its established role during vascular morphogenesis as a regulator of rapid vascular responses. Here we show that mice deficient in Ang-2 (encoded by the gene Angpt2) cannot elicit an inflammatory response in thioglycollate-induced or Staphylococcus aureus-induced peritonitis, or in the dorsal skinfold chamber model. Recombinant Ang-2 restores the inflammation defect in Angpt2(-/-) mice. Intravital microscopy showed normal TNF-alpha-induced leukocyte rolling in the vasculature of Angpt2(-/-)mice, but rolling cells did not firmly adhere to activated endothelium. Cellular experiments showed that Ang-2 promotes adhesion by sensitizing endothelial cells toward TNF-alpha and modulating TNF-alpha-induced expression of endothelial cell adhesion molecules. Together, these findings identify Ang-2 as an autocrine regulator of endothelial cell inflammatory responses. Ang-2 thereby acts as a switch of vascular responsiveness exerting a permissive role for the activities of proinflammatory cytokines.