Involvement of Ca2+/calmodulin-dependent protein kinase II in endothelial NO production and endothelium-dependent relaxation

Nitric oxide (NO) is synthesized from l-arginine by the Ca(2+)/calmodulin-sensitive endothelial NO synthase (NOS) isoform (eNOS). The present study assesses the role of Ca(2+)/calmodulin-dependent protein kinase II (CaMK II) in endothelium-dependent relaxation and NO synthesis. The effects of three CaMK II inhibitors were investigated in endothelium-intact aortic rings of normotensive rats. NO synthesis was assessed by a NO sensor and chemiluminescence in culture medium of cultured porcine aortic endothelial cells stimulated with the Ca(2+) ionophore A23187 and thapsigargin. Rat aortic endothelial NOS activity was measured by the conversion of l-[(3)H]arginine to l-[(3)H]citrulline. Three CaMK II inhibitors, polypeptide 281-302, KN-93, and lavendustin C, attenuated the endothelium-dependent relaxation of endothelium-intact rat aortic rings in response to acetylcholine, A23187, and thapsigargin. None of the CaMK II inhibitors affected the relaxation induced by NO donors. In a porcine aortic endothelial cell line, KN-93 decreased NO synthesis and caused a rightward shift of the concentration-response curves to A23187 and thapsigargin. In rat aortic endothelial cells, KN-93 significantly decreased bradykinin-induced eNOS activity. These results suggest that CaMK II was involved in NO synthesis as a result of Ca(2+)-dependent activation of eNOS.     

Olfactory receptors on the maxillary palps of small ermine moth larvae: evolutionary history of benzaldehyde sensitivity

In lepidopterous larvae the maxillary palps contain a large portion of the sensory equipment of the insect. Yet, knowledge about the sensitivity of these cells is limited. In this paper a morphological, behavioral, and electrophysiological investigation of the maxillary palps of Yponomeuta cagnagellus (Lepidoptera: Yponomeutidae) is presented. In addition to thermoreceptors, CO₂ receptors, and gustatory receptors, evidence is reported for the existence of two groups of receptor cells sensitive to plant volatiles. Cells that are mainly sensitive to (E)-2-hexenal and hexanal or to (Z)-3-hexen-1-ol and 1-hexanol were found. Interestingly, a high sensitivity for benzaldehyde was also found. This compound is not known to be present in Euonymus europaeus, the host plant of the monophagous Yponomeuta cagnagellus, but it is a prominent compound in Rosaceae, the presumed hosts of the ancestors of Y. cagnagellus. To elucidate the evolutionary history of this sensitivity, and its possible role in host shifts, feeding responses of three Yponomeuta species to benzaldehyde were investigated. The results confirm the hypothesis that the sensitivity to benzaldehyde evolved during the ancestral shift from Celastraceae to Rosaceae and can be considered an evolutionary relict, retained in the recently backshifted Celastraceae-specialist Y. cagnagellus.     

Evolution of gustatory sensitivity in Yponomeuta caterpillars: sensitivity to the stereo-isomers dulcitol and sorbitol is localised in a single sensory cell

The cellular localisation of the sensitivity to two host-plant specific larval phagostimulants was determined in Yponomeuta evonymellus and in inter-specific hybrids of Y. cagnagellus and Y. padellus. The combined results of cross-adaptation experiments, analysis of mixture responses and inter-spike interval distributions indicated that the sensitivity for sorbitol and dulcitol is localised in the same cell in the investigated species. This result suggests that during the evolution of the genus the main alteration in the mechanism of sugar-alcohol detection occurred at the level of the receptor proteins. Relatively simple modification of this kind can, however, have large effects, since larvae with modified receptor proteins will directly respond to the new stimulus while all central processing and resulting behaviour can remain unmodified. Accepted: 7 October 1998     

Sympatric speciation in Yponomeuta: no evidence for host plant fidelity

According to sympatric speciation theory, adaptation to different host plants is expected to pleiotropically lead to assortative mating, an important factor in the reduction of gene flow between the diverging subpopulations. This scenario predicts mating on and oviposition preference for the respective hosts in both the diverging subpopulations and recently originated species. Here, we test both predictions in the oligophagous Yponomeuta padellus (L.) and the monophagous Yponomeuta cagnagellus (Hübner) (Lepidoptera: Yponomeutidae), two closely related small ermine moth species from the western European clade of Yponomeuta for which speciation in sympatry has been proposed. Mating location and adult host acceptance were evaluated under both semi-field (in a large outdoor cage with a choice of host and non-host plants) and field conditions. In the semi-field experiment, only Y. cagnagellus showed some preference for mating on its own host (16% of all mating pairs) over non-host plants (3% of all mating pairs). However, in both species, more than 80% of the mating pairs were not formed on a plant but instead on the cage itself. Further examination of the mating site of Y. cagnagellus in the field revealed no preference for host plants over non-host plants in the two consecutive years of observation. Yponomeuta padellus females, collected from and reared on Prunus spinosa L. (Rosaceae), showed an oviposition preference for the alternative host Crataegus monogyna Jacq. (Rosaceae) in the semi-field experiment. We thus found no evidence that host-plant fidelity (in terms of mating site) has been the driving force in the speciation process of these Yponomeuta species, nor did we find evidence of host race formation in the tested population of the oligophagous Y. padellus .     

Physiological factors influencing preference for wet and dry food in Schistocerca gregaria nymphs

The physiological factors responsible for behavioral regulation of water balance by differential feeding in Schistocerca gregaria were investigated. Locusts given the option to regulate their water intake by feeding on high-water content (wet) food or low-water content (dry) food had values for percent body water and hemolymph volumes intermediate between those for locusts given either food exclusively. Locusts given wet food only prior to no-choice tests had relatively small meals and long latencies to feed on wet food and large meals and short latencies to feed on dry food. Increasing volume by adding 75 l normal saline did not alter meal size on either food, while locusts given dry food prior to testing had the opposite behavior. Increasing hemolymph concentration led to smaller meals on both wet and dry food, but diluting the hemolymph by an equal volume of distilled water resulted in larger meals on dry food and smaller meals on wet food. Removing an equal volume of hemolymph had no effect, but combining hemolymph removal with increased osmotic pressure reduced the meal size on dry food and increased meal size on wet food.

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Résumé L'étude a porté sur les bases physiologiques de la régulation comportementale de l'équilibre hydrique par alimentation différentielle chez S. gregaria. Les teneurs en eau (pourcentage du poids du corps) et les volumes d'hémolymphe des criquets ayant eu la possibilité de moduler leur apport en eau en conssommant des aliments à forte teneur en eau (frais) ou à faible teneur (secs), étaient intermédiaires entre ceux des criquets ayant l'un ou l'autre exclusivement. Les criquets, ayant uniquement des aliments frais avant des expériences sans choix, ont présenté des temps de latence prolongés et des repas relativement brefs sur aliments frais, et des temps de latence brefs et des repas abondants sur aliments secs; les résultats ont été inversés quand l'alimentation préalable était sur nourriture sèche.Ces résultats sont partiellement simulés par des altérations du volume et de la pression osmotique de l'hémolymphe. L'augmentation de la concentration de l'hémolymphe entraîne des repas moins abondants sur aliments frais et secs, vraisemblablement par suite de signaux de satiété. L'augmentation du volume en ajoutant 75 l de solution physiologique n'a pas modifié la taille du repas sur les deux types d'aliments, mais la dilution de l'hémolymphe par un volume égal d'eau distillée a induit des repas plus importants sur aliment sec, et plus petit sur aliment frais. Une augmentation de volume plus importante, de 150 l, réduisait la taille du repas sur aliment frais, mais pas sur aliment sec. Le prélèvement d'environ 75 l d'hémolymphe a été sans effet; mais, combiné avec l'augmentation de la pression osmotique, il a réduit la taille du repas sur aliment sec (comme l'augmentation de la pression osmotique seule), mais a augmenté la taille du repas sur aliment frais. Ce dernier résultat est semblable à ceux obtenus lors des études du déterminisme de l'abreuvement. La discussion porte sur la relation entre le contrôle de l'absorption de l'eau et des aliments.

     

The Relationship Between Tool Use and Prey Availability in Chimpanzees (Pan troglodytes schweinfurthii) of Northern Democratic Republic of Congo

International Journal of Primatology - A key feature of human behavioral diversity is that it can be constrained by cultural preference (“cultural override”); that is,...     

Correction to: The Relationship Between Tool Use and Prey Availability in Chimpanzees (Pan troglodytes schweinfurthii) of Northern Democratic Republic of Congo

International Journal of Primatology - The original version of this article unfortunately contained mistakes in the Discussion and Acknowledgments sections. The changes are shown below:     

Oviposition and tarsal chemoreceptors of the cabbage root fly are stimulated by glucosinolates and host plant extracts

The role of glucosinolates in the oviposition behaviour of the cabbage root fly,Delia radicum (L.) (Diptera, Anthomyiidae) was investigated using egg counts and electrophysiological recordings from tarsal contact chemoreceptors. The glucosinolates present both inside and on the surface of cauliflower leaves were determined. The total amounts obtained with the two methods differed by a factor of 100. The extract of the leaf surface contained about 60 μg per g leaf extracted (gle), the total leaf extract 7.5 mg per gle. The glucosinolate patterns of the two extracts were qualitatively similar, but the ratios of the content of individual glucosinolates showed considerable differences. The D sensilla on segment 3 and 4 of the tarsus ofD. radicum females were shown to contain a sensitive receptor cell for glucosinolates. In contrast, the receptor cells of the D sensilla of the other segments did not respond in a dose dependent way to these compounds. The glucosinolate receptors were found to be especially sensitive to glucobrassicin, gluconasturtiin and glucobrassicanapin with thresholds of about 10⁻⁸ M to 10⁻⁹ M. Large differences (up to two orders of magnitude) were observed among the different glucosinolates. A significant correlation was found between the behavioural discrimination index and the electrophysiological results. But no obvious correlation existed between the chemical nature of the glucosinolate side chain (e.g. indole, aromatic and aliphatic groups), and their stimulatory activity. However, a significant correlation was found between the overall length of the side chain and the biological activity. Although the flies discriminated clearly between model leaves with and without glucosinolates, a clear dose response curve was only obtained for the indole glucosinolate glucobrassicin. Since the most stimulatory fraction of the surface extract contained no glucosinolates, it was concluded that other compounds, in addition to glucosinolates, do play an important role for the stimulation of oviposition.     

Perception of the oviposition deterring pheromone by tarsal and abdominal contact chemoreceptors in Pieris brassicae

Perception of the oviposition deterring pheromone by contact chemoreceptors in female Pieris brassicae was studied employing a tip recording technique. Electrophysiological responses of tarsal taste hairs to eggwash solutions show a marked increase in frequency of spikes originating from only one sensory cell. This suggests that in foretarsal taste hairs females, apart from the glucosinolate cells also possess sense cells specifically sensitive to the oviposition deterring pheromone.Morphological studies by means of the scanning electron microscope revealed that the ovipositor of P. brassicae is provided with two groups of contact chemoreceptors. Electrophysiological recordings from these sensilla indicate the presence of at least three sensory cells, one of them being a mechanoreceptor. Stimulation with eggwash evokes a slight increase in spike frequency which cannot be ascribed to one particular sense cell. This indicates that abdominal taste hairs in some way may participate in the perception of the oviposition deterring pheromone. Responses to glucosinolates do not differ significantly from control stimulations.

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Résumé Une technique d'enregistrement apical a été utilisée pour examiner la perception d'une phéromone dissuadant la ponte par les poils récepteurs chimiques de contact des femelles de Pieris brassicae. Les réponses électrophysiologiques des poils gustatifs des tarses en présence de solutions de rinçage d'oeufs présentent une fréquence marquée des potentiels d'action provenant principalement d'une cellule sensorielle. Ceci suggère que les poils gustatifs des tarses des pattes antérieures des femelles possèdent, en plus de cellules répondant aux glucosinolates, des cellules sensorielles sensibles spécialement à la phéromone dissuadant la ponte.Des études morphologiques au microscope à balayage révèlent que l'oviposition de P. brassicae est pourvu de deux groupes de chimiorécepteurs de contact. Des enregistrements électrophysiologiques de ces sensilles révèlent la présence d'au moins trois cellules sensorielles, l'une d'entre elles étant un mécanorécepteur. La stimulation avec la solution de rinçage des oeufs évoque un léger accroissement de la fréquence des potentiels d'action qui ne peut être attribué à une cellule sensorielle particulière. Ceci indique que les poils gustatifs abdominaux peuvent participer d'une certaine façon à la perception de la phéromone dissuadant la ponte. Les réponses aux glucosinolates ne diffèrent pas significativement des stimulations témoin.

     

Pollination of Specklinia by nectar-feeding Drosophila: the first reported case of a deceptive syndrome employing aggregation pheromones in Orchidaceae

Background and Aims The first documented observation of pollination in Pleurothallidinae was that of Endrés, who noticed that the ‘viscid sepals’ of Specklinia endotrachys were visited by a ‘small fly’. Chase would later identify the visiting flies as being members of the genus Drosophila. This study documents and describes how species of the S. endotrachys complex are pollinated by different Drosophila species.Methods Specimens of Specklinia and Drosophila were collected in the field in Costa Rica and preserved in the JBL and L herbaria. Flies were photographed, filmed and observed for several days during a 2-year period and were identified by a combination of non-invasive DNA barcoding and anatomical surveys. Tissue samples of the sepals, petals and labellum of Specklinia species were observed and documented by SEM, LM and TEM. Electroantennogram experiments were carried out on Drosophila hydei using the known aggregation pheromones ethyl tiglate, methyl tiglate and isopropyl tiglate. Floral compounds were analysed by gas chromatography–mass spectometry using those same pheromones as standards.Key Results Flowers of S. endotrachys, S. pfavii, S. remotiflora and S. spectabilis are visited and pollinated by several different but closely related Drosophila species. The flies are arrested by aggregation pheromones, including ethyl tiglate, methyl tiglate and isopropyl tiglate, released by the flowers, and to which at least D. hydei is very sensitive. Visible nectar drops on the adaxial surface of sepals are secreted by nectar-secreting stomata, encouraging male and female Drosophila to linger on the flowers for several hours at a time. The flies frequently show courtship behaviour, occasionally copulating. Several different Drosophila species can be found on a single Specklinia species.Conclusions Species of the S. endotrachys group share a similar pollination syndrome. There seem to be no species-specific relationships between the orchids and the flies. It is not expected that Specklinia species will hybridize naturally as their populations do not overlap geographically. The combination of pheromone attraction and nectar feeding is likely to be a generalized pollination syndrome in Pleurothallidinae.