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Solubilization and pharmacological characterization of a glucocorticoid membrane receptor from an amphibian brain.

Physiological functions of steroid hormones involve activation of intracellular receptors as well as poorly understood membrane receptors. We report the pharmacological characterization of a solubilized corticosterone receptor from neuronal membranes. This receptor previously was shown to localize with plasma membrane subcellular fractions and to be involved in the modulation of courtship behaviors in the roughskin newt (Taricha granulosa). We describe procedures with non-ionic detergents that solubilize the receptor and maintain high affinity [3H]corticosterone binding. The pharmacology of the solubilized corticosterone receptor resembles that of the membrane receptor with high affinity for [3H]corticosterone and an identical rank-order potency for other steroid ligands (corticosterone>cortisol>aldosterone>dexamethasone). Unlike binding in membrane preparations, [3H]corticosterone binding to the solubilized receptor is insensitive to negative modulation by guanyl nucleotides and only modestly sensitive to the presence of Mg2+. We also identified two ligands that exhibit high affinity binding to the solubilized receptor and have the potential to be used in an affinity purification scheme. They are corticosterone-3-carboxymethyloxime (CORT-3-CMO), which may be covalently attached to a Sepharose resin, and a derivitized azide form of CORT-3-CMO which can be covalently coupled to the solubilized receptor itself. The stability of the solubilized [3H]corticosterone receptor in the detergent system will facilitate further purification and molecular characterization.

Affinity Labels↗

Temporal mating isolation driven by a behavioral gene in Drosophila.

Speciation is the evolutionary process in which new barriers to gene exchange are created. These barriers may be physical, leading to spatial separation of subpopulations and resulting in allopatric speciation, or they may be temporal, giving rise to allochronic speciation, and may include the time of day or the time of year when mating takes place. Drosophila melanogaster and D. pseudoobscura show different temporal patterns of circadian locomotor activity that are determined by the circadian clock gene period (per). Genes that control aspects of behavior that might be relevant to courtship and mating, such as locomotor patterns, become obvious candidates for involvement in the speciation process. However, evidence for the role of individual genes in the mechanism of mate choice has proved elusive. We have used transgenic flies carrying the natural per genes from these two Drosophila species to reveal that per has the potential to provide the permissive conditions for speciation, by affecting mate choice through a mechanism involving the species-specific timing of mating behavior.

Animals↗

Daily variations of antioxidant enzyme and luciferase activities in the luminescent click-beetle Pyrearinus termitilluminans: cooperation against oxygen toxicity.

Several lines of investigation have suggested an interplay between bioluminescence (BL) and oxyradical metabolism, mainly in bacteria and beetles. Although not yet confirmed, luminescent beetles seem to be challenged daily by oxidative conditions imposed by higher oxygen absorption necessary to enhance light emission for courtship (adult lampyrids and elaterids) and prey attraction (e.g. Pyrearinus termitilluminans larvae). This work reports the activities of luciferase, superoxide dismutase (SOD), catalase and dehydroascorbate reductase (DHAR) and total glutathione content at different times of the day in the bright prothorax and dim abdomen of larval Pyrearinus termitilluminans (Coleoptera: Elateridae), investigating a possible adjuvant role for luciferase in oxygen detoxification. Luciferase activity in the prothorax was shown to peak at 7 p.m., which is the time when P. termitilluminans larvae light up for prey attraction. In their habitat, P. termitilluminans larvae emit light until 8.30 p.m. However, at 8 p.m., prothorax luciferase activity achieved basal levels and total glutathione content declined to the daily lowest value, possibly resulting from hyperoxidative conditions during this time. Significant increases in the activities of total SOD (28%) and catalase (37%) were observed in the prothorax at 9 p.m., which should minimize the extent of damage from this potentially hazardous period. Prothorax total SOD (42% higher than daily average) and abdomen CuZnSOD (41%) and catalase (95%) activities showed extra peaks at 7-10 a.m., and abdomen DHAR activity was maximal (37%) earlier (4-7 a.m.). These morning increases in antioxidant enzyme activities may be associated with biological events other than bioluminescence, e.g. intense physical activity for digging tunnels and/or digestion of captured preys. These data suggest that oxyradical pathway and bioluminescence are coordinated, especially in the prothorax, to minimize the oxidative stress imposed by higher irrigation of the photocytes with O(2) when P. termitilluminans larvae emit light.

Animals↗

Phylogeny of Tetraoninae and other galliform birds using mitochondrial 12S and ND2 genes.

The avian subfamily Tetraoninae (grouse and ptarmigan) is a Holarctic group in the order Galliformes distinguished by morphological adaptations to cold environments and behavioral traits associated with elaborate courtship. Here we investigate the relationships of 17 tetraonines and 12 other galliform species using mitochondrial 12S and ND2 sequence data. We found support for the recent phylogenetic classification that separates the genus Dendragapus into two genera, Falcipennis and Dendragapus. In addition, we found support for a tetraonine clade in which the first divergence is between Bonasa umbellus and all others, followed by divergence between a Bonasa bonasia/Bonasa sewerzowi clade and the remaining tetraonines. Falcipennis canadensis is sister to a clade with four Tetrao species, and the genus Centrocercus is sister to a Dendragapus obscurus/Tympanuchus clade. Our data indicate a basal position for Cracidae and Megapodiidae among the five recognized galliform families. We also found strong support for the monophyly of Phasianidae, although the relative positions of Numididae and Odontiphoridae remains unresolved. We use a maximum likelihood approach to infer ages of 37mya for divergence of Numididae and Phasianidae and 28mya for the divergence of Tetraoninae and Meleagris gallopavo. These estimates must be viewed as tentative as they depend on tests of rates of molecular evolution and accurate fossil dates.

Animals↗

Body mass, age and sexual maturity in short-beaked echidnas, Tachyglossus aculeatus.

During the course of this 12 year field study body masses of 11 hatchling echidnas (Tachyglossus aculeatus multiaculeatus) and 25 pouch young between the ages of 5 and 60 days were recorded. Body mass increased from 0.3 to approximately 50 g in the first half of pouch life. It then quadrupled before young were placed in a burrow at 45 to 55 days of age. There was a positive correlation between the body mass of the female and that of her young at weaning. From 33 subadult echidnas located, tagged and radio tracked during this study, body masses of 10 were monitored to sexual maturity, i.e. when first encountered in a courtship train. Minimum age of sexual maturity ranged between 5 and 12 years. As subadults, there was no difference between mean body masses of males and females. At sexual maturity, mean body mass of females was significantly higher. No correlation was found between age at sexual maturity and body mass nor was there a significant difference in age of males and females at sexual maturity.

Animals↗

The three steroidal components of the goldfish preovulatory pheromone signal evoke different behaviors in males.

The goldfish sex pheromone system is the best understood among the teleost fishes. Pheromones in this species are unspecialized hormonal products, which are released in ratios that vary with reproductive status. This study examined behavioral responses of male goldfish to three steroidal components of the female preovulatory pheromone: 17,20beta-dihydroxy-4-pregnen-3-one (1720betaP); 17,20beta-dihydroxy-4-pregnen-3-one-20-sulfate (1720betaP-S); and androstenedione (AD). Males were observed during exposure to nanomolar concentrations of each steroid over a 2-h period. We observed chasing, nudging (courtship behaviors) and pushing (an aggressive behavior). Each steroid elicited a different set of behaviors. 1720betaP, which is released by ovulatory females, elicited a low level of chasing and nudging that persisted throughout the experiment. Exposure to 1720betaP-S, which is released primarily by ovulatory females, triggered a large increase in nudging and chasing that lasted for only 5 min. In contrast, AD, which is released by females early in the ovulatory cycle and by mature males, elicited increases in aggressive behavior. 1720betaP and 1720betaP-S both caused increases in GtH-II release while AD did not. These results demonstrate that goldfish can discriminate components found in the female pheromone blend, suggesting that goldfish, and likely other fish species, may employ blends of hormonal products as pheromones.

Aggression↗

Mating interactions between two biotypes of the whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae) in Australia.

The biological consequences of mating interactions between indigenous and exotic biotypes of Bemisia tabaci (Gennadius) in Australia were studied using a combination of field and laboratory experiments. The key results of the interaction between the B and eastern Australian biotypes were reduced population increase, a marked increase in the proportion of male progeny, fewer eggs produced by females paired with males of different biotype and no difference in the numbers of eggs per unmated female and females paired with males of the same biotype. In addition, there was no change in the proportion of eggs hatching, mixed biotype pairs spent more time courting than single biotype pairs and a low level of hybridization in field cages and small containers was observed. These observations suggest three possibilities. The first is the 'distracting male hypothesis' in which mating pairs made up of different biotypes apportion more time to courtship and less time to egg laying than single biotype pairs. The second invokes the 'single-locus complementary sex determination model' in which the production of non-viable diploid male zygotes may explain the reduction in eggs laid. The third is cytoplasmic incompatibility between biotypes caused by Wolbachia. The results also suggest that the geographical distribution of clusters of related biotypes both overseas and in Australia may be explained by between-biotype interactions leading to the formation of parapatric populations.

Animals↗

Genetic analysis of Drosophila virilis sex pheromone: genetic mapping of the locus producing Z-(11)-pentacosene.

Z-(11)-pentacosene, Drosophila virilis sex pheromone, is predominant among the female cuticular hydrocarbons and can elicit male courtship behaviours. To evaluate the genetic basis of its production, interspecific crosses between D. novamexicana and genetically marked D. virilis were made and hydrocarbon profiles of their backcross progeny were analysed. The production of Z-(11)-pentacosene was autosomally controlled and was recessive. Of the six D. virilis chromosomes only the second and the third chromosomes showed significant contributions to sex pheromone production, and acted additively. Analysis of recombinant females indicated that the locus on the second chromosome mapped to the proximity of position 2-218.

Alkenes↗

Genetic analysis of an intersex allele (ix5) that regulates sexual phenotype of both female and male Drosophila melanogaster.

An allele of intersex (ix5) of Drosophila melanogaster has been characterized. The genetic analysis of the allele demonstrated that like other point mutations of ix, the ix5 allele also transformed diplo-X individuals into intersexes. The ix5 mutation also affects the arrangement of sex comb bristles on the forelegs of males, although they had morphologically nearly normal male genitalia. They often fail to display a sustained pattern of courtship activity when tested. Orcein-stained squash preparations of testes from ix5 males revealed a defect in spermatogenesis. Our results, taken together with those of McRobert & Tompkins (1985), indicate that the ix+ gene also functions in male sex determination.

Alleles↗

Evidence for assortative mating on digit ratio (2D:4D), a biomarker for prenatal androgen exposure.

The second-to-fourth digit ratio (2D:4D) presents an anatomical sex difference in humans. On average, men tend to have lower 2D:4D compared with women. There is fairly strong evidence for a role of the 2D:4D ratio as a biomarker for the organizational (permanent) effects of prenatal testosterone on the brain and behaviour. Recently, an accumulating research programme has shown 2D:4D to be related to a multitude of sex-dependent, hormonally influenced biosocial traits and phenotypes which reach into the domains of ability, behaviour, fertility, health, personality and sexuality. This study investigated the degree of assortative mating (spousal similarity) in a sample of 239 native Austrian couples of parental or grandparental age, all of them having reproduced. Results included: (i) significant spousal correlations of +0.19 and +0.18 for right-hand and left-hand 2D:4D, respectively, and +0.24 for average 2D:4D; (ii) no assortative mating effect on the right-minus-left difference in 2D:4D; (iii) indications consistent with a possible generational decrease of spousal similarity in 2D:4D; (iv) a prevalence of couples with a lower right-hand 2D:4D observed in the husband compared with his wife; and (v) relations of spousal 2D:4D patterns to spousal age differences, such that matings of men with more male-typical trait expressions (namely, a generally low right-hand 2D:4D or showing a lower right-minus-left 2D:4D difference than their wives) implicated larger male-minus-female age differences, i.e. younger wives. It is argued that assortative mating on 2D:4D operates indirectly and may be mediated through the assortment on other, more perceptible, physical traits and psychological phenotypes that entertain associations with 2D:4D and are relevant for courtship and mate choice.

Adult↗

Language and life history: a new perspective on the development and evolution of human language.

It has long been claimed that Homo sapiens is the only species that has language, but only recently has it been recognized that humans also have an unusual pattern of growth and development. Social mammals have two stages of pre-adult development: infancy and juvenility. Humans have two additional prolonged and pronounced life history stages: childhood, an interval of four years extending between infancy and the juvenile period that follows, and adolescence, a stage of about eight years that stretches from juvenility to adulthood. We begin by reviewing the primary biological and linguistic changes occurring in each of the four pre-adult ontogenetic stages in human life history. Then we attempt to trace the evolution of childhood and juvenility in our hominin ancestors. We propose that several different forms of selection applied in infancy and childhood; and that, in adolescence, elaborated vocal behaviors played a role in courtship and intrasexual competition, enhancing fitness and ultimately integrating performative and pragmatic skills with linguistic knowledge in a broad faculty of language. A theoretical consequence of our proposal is that fossil evidence of the uniquely human stages may be used, with other findings, to date the emergence of language. If important aspects of language cannot appear until sexual maturity, as we propose, then a second consequence is that the development of language requires the whole of modern human ontogeny. Our life history model thus offers new ways of investigating, and thinking about, the evolution, development, and ultimately the nature of human language.

Adolescent↗

The evolution of male mate choice in insects: a synthesis of ideas and evidence.

Mate choice by males has been recognized at least since Darwin's time, but its phylogenetic distribution and effect on the evolution of female phenotypes remain poorly known. Moreover, the relative importance of factors thought to underlie the evolution of male mate choice (especially parental investment and mate quality variance) is still unresolved. Here I synthesize the empirical evidence and theory pertaining to the evolution of male mate choice and sex role reversal in insects, and examine the potential for male mating preferences to generate sexual selection on female phenotypes. Although male mate choice has received relatively little empirical study, the available evidence suggests that it is widespread among insects (and other animals). In addition to 'precopulatory' male mate choice, some insects exhibit 'cryptic' male mate choice, varying the amount of resources allocated to mating on the basis of female mate quality. As predicted by theory, the most commonly observed male mating preferences are those that tend to maximize a male's expected fertilization success from each mating. Such preferences tend to favour female phenotypes associated with high fecundity or reduced sperm competition intensity. Among insect species there is wide variation in mechanisms used by males to assess female mate quality, some of which (e.g. probing, antennating or repeatedly mounting the female) may be difficult to distinguish from copulatory courtship. According to theory, selection for male choosiness is an increasing function of mate quality variance and those reproductive costs that reduce, with each mating, the number of subsequent matings that a male can perform ('mating investment') Conversely, choosiness is constrained by the costs of mate search and assessment, in combination with the accuracy of assessment of potential mates and of the distribution of mate qualities. Stronger selection for male choosiness may also be expected in systems where female fitness increases with each copulation than in systems where female fitness peaks at a small number of matings. This theoretical framework is consistent with most of the empirical evidence. Furthermore, a variety of observed male mating preferences have the potential to exert sexual selection on female phenotypes. However, because male insects typically choose females based on phenotypic indicators of fecundity such as body size, and these are usually amenable to direct visual or tactile assessment, male mate choice often tends to reinforce stronger vectors of fecundity or viability selection, and seldom results in the evolution of female display traits. Research on orthopterans has shown that complete sex role reversal (i.e. males choosy, females competitive) can occur when male parental investment limits female fecundity and reduces the potential rate of reproduction of males sufficiently to produce a female-biased operational sex ratio. By contrast, many systems exhibiting partial sex role reversal (i.e. males choosy and competitive) are not associated with elevated levels of male parental investment, reduced male reproductive rates, or reduced male bias in the operational sex ratio. Instead, large female mate quality variance resulting from factors such as strong last-male sperm precedence or large variance in female fecundity may select for both male choosiness and competitiveness in such systems. Thus, partial and complete sex role reversal do not merely represent different points along a continuum of increasing male parental investment, but may evolve via different evolutionary pathways.

Animals↗

Close-range attraction in Lygocoris pabulinus (L.).

Males of the green capsid bug, Lygocoris pabulinus, exhibit a specific courtship behavior, i.e., a vibration of the abdomen. When both live and dead females were offered to males, this vibration behavior was elicited in most of the males tested. When females were dissected into separate body parts, heads, wings, and legs elicited equal responses, while thorax plus abdomen elicited a much lower response. When separate body parts were extracted, the leg extracts elicited significantly stronger responses than any other extract. This suggests that female L. pabulinus legs are either the source of a close-range sex pheromone or that pheromone is accumulated on the legs due to grooming behavior. The leg extracts contained several hydrocarbons such as n-alkenes, n-alkanes, and some methylalkanes. Female extracts contained more (Z)-9-pentacosene and male extracts contained more (Z)-9-heptacosene. Substrates on which females had walked elicited similar responses as female legs, indicating that the pheromone is deposited on the substrate. This enlarges the functional range of low-volatility compounds, which are thought to function only when sexes are in close vicinity or in contact.

Abdomen↗

Identification and synthesis of a male-produced sex pheromone from the stink bug Chlorochroa sayi.

The reproductive behavior of the stink bug Chlorochroa sayi was studied in the laboratory. There was a sexual maturation period of about 10 days before bugs began mating. Sexually mature adult bugs engaged in courtship consisting of antennation and head-butting of the female by the male, before the female adopted a receptive posture and copulation occurred. Both sexes mated multiple times during their life-spans, with the mean duration of copulations of virgin bugs (42.3 +/- 19.6 min) and experienced bugs (37.3 +/- 28.4 min) being similar. Most matings were initiated in the late afternoon or evening, when pheromone production by males was greatest. Males transferred sperm and nutrients constituting about 17% of their body weight to females during mating. Three male-specific components, methyl geranate, methyl citronellate, and methyl (E)-6-2,3-dihydrofarnesoate in a ratio of 100:0.45:1.6, were first detected in volatiles collected from male bugs on green beans about 9-12 days after the final molt to the adult stage. In vertical Y-tube bioassays, females were attracted to odors from mature male bugs, and to a blend of the three male-produced components. Low numbers of females also were attracted in field trials with the three-component blend. The relatively weak attraction may be a result of other, as yet unknown cues being required in addition to the pheromone, such as visual or substrate-borne vibrational cues.

Animals↗

(Z)-3-hexenyl (R)-3-hydroxybutanoate: a male specific compound in three North American decorator wasps Eucerceris rubripes, E. conata and E. tricolor.

The males of the decorator wasps Eucerceris have been observed to display abdomen-dragging behavior on plants surrounding their nest. It is thought that this applies a territorial-marking sex pheromone that serves to alert females to the males' territory for courtship and mating. The extracts of three species E. rubripes, E. conata, and E. tricolor have been analyzed by GC-MS. The gas chromatograms revealed the presence, in large quantities, of one common volatile compound in the male head extracts. By analogy with the closely related bee wolves Philanthus, we believe the pheromone is produced in the mandibular glands. Utilizing mass spectrometry, microreactions, FT-IR, and 1H and 13C NMR spectroscopy the structure of the pheromone was proposed to be (Z)-3-hexenyl 3-hydroxybutanoate, which was confirmed by synthesis. The absolute configuration of the chiral center was determined to be R for the three species by preparing the Mosher esters of the insect samples and comparing their GC retention times with a synthetic sample of known absolute configuration. In addition 2- and 3-hexenoic acid and some aromatic compounds were also found in varying quantities in both males and females along with hydrocarbons and fatty acids, although no species-specific profiles emerged.

Animals↗

Evidence for a sex pheromone in bark beetle parasitoid Roptrocerus xylophagorum.

Male Roptrocerus xylophagorum (Ratzeburg) (Hymenoptera: Pteromalidae) exhibited courtship and mating behaviors including wing fanning, antennation, mounting, and copulation attempts when exposed to glass bulb decoys coated with a whole-body extract of females in hexane, acetone, or methanol. Activity of extract-treated decoys declined gradually over one week. Males responded much less strongly to freeze-killed female cadavers extracted with solvents than to unextracted cadavers; treatment of extracted cadavers with female extract restored male responses. The pheromone was found to be equally present over the surface of both the abdomen and head/thorax of females, and the origin of the pheromone could not be conclusively localized to any single body region. The activity of pheromone on females increased between day 1 and days 3-5 following eclosion; otherwise, pheromone activity was not significantly affected by either female age or mating. Males were arrested within the zone of a glass surface on which females had walked, suggesting that the pheromone might be substrate-borne. Recent exposure to females reduced male responsiveness, but responsiveness was fully restored after a few hours of male isolation from females. When hexane extracts of whole females were fractionated on silica gel, the pheromone's activity was largely recovered with the first, most nonpolar fraction. Female extracts and fractions were analyzed by coupled gas chromatography-mass spectrometry. Cuticular hydrocarbon alkanes were identified as the extract components whose concentrations correlated best with male responses. Evidence of the pheromone's long persistence, low volatility, low polarity, and presence over the insect's entire body surface further supported the hypothesis that the pheromone was composed of one or more cuticular hydrocarbons.

Animals↗

Oligosaccharide composition and pheromonal activity of male tergal gland secretions of the German cockroach, Blattella gennanica (L.).

In the sequential courtship behavior of the German cockroach, Blattella germanica (Dictyoptera: Blattellidae), females feed on the tergal gland secretions from the male tergites. The phagostimulative components in the male secretions were composed of a complex mixture of oligosaccharides and phospholipids. Besides seven oligosaccharides previously identified, two new trisaccharides, O-alpha-D-glucopyranosyl-(1 --> 6)-alpha-D-glucopyranosyl alpha-D-glucopyranoside and O-alpha-glucopyranosyl-(1 --> 4)-O-alpha-glucopyranosyl-(1 --> 1)-myo-inositol, were characterized as the sugar components of the secretions. The activity of the sugar components was compared in combination with the lipid fraction. Individual oligosaccharides exhibited the activity either alone or synergistically with the phospholipids. A complex mixture of these primary substances in the tergal gland secretions, thus, serves as a pheromonal phagostimulant in the precopulatory behavior, strongly eliciting feeding response in the female cockroach.

Animals↗

Trans-bergamotenes-male pheromone of the ectoparasitoid Melittobia digitata.

The first male-produced sex attractant pheromone in the parasitic Hymenoptera has been identified. The elaborate courtship behavior of Melittobia digitata, an idiobiont that parasitizes the last larval instar or the pupal stage of solitary bees and wasps, involves a series of coordinated movements of legs, wings, and antennae, initiated after the female is attracted to the blind, flightless male. We identified alpha- and beta-trans-bergamotene as the active compounds of the male M. digitata sex attractant. Variation in the release of the sex pheromone by males and the pheromone load during aging is also described.

Animals↗