Cuticular Hydrocarbons Production Research Articles

Dissection of Oenocytes from Adult <em>Drosophila melanogaster</em>

In Drosophila melanogaster, as in other insects, a waxy layer on the outer surface of the cuticle, composed primarily of hydrocarbon compounds, provides protection against desiccation and other environmental challenges. Several of these cuticular hydrocarbon (CHC) compounds also function as semiochemical signals, and as such mediate pheromonal communications between members of the same species, or in some instances between different species, and influence behavior. Specialized cells referred to as oenocytes are regarded as the primary site for CHC synthesis. However, relatively little is known regarding the involvement of the oenocytes in the regulation of the biosynthetic, transport, and deposition pathways contributing to CHC output. Given the significant role that CHCs play in several aspects of insect biology, including chemical communication, desiccation resistance, and immunity, it is important to gain a greater understanding of the molecular and genetic regulation of CHC production within these specialized cells. The adult oenocytes of D. melanogaster are located within the abdominal integument, and are metamerically arrayed in ribbon-like clusters radiating along the inner cuticular surface of each abdominal segment. In this video article we demonstrate a dissection technique used for the preparation of oenocytes from adult D. melanogaster. Specifically, we provide a detailed step-by-step demonstration of (1) how to fillet prepare an adult Drosophila abdomen, (2) how to identify the oenocytes and discern them from other tissues, and (3) how to remove intact oenocyte clusters from the abdominal integument. A brief experimental illustration of how this preparation can be used to examine the expression of genes involved in hydrocarbon synthesis is included. The dissected preparation demonstrated herein will allow for the detailed molecular and genetic analysis of oenocyte function in the adult fruit fly.

Dissection of Oenocytes from Adult <em>Drosophila melanogaster</em>

In Drosophila melanogaster, as in other insects, a waxy layer on the outer surface of the cuticle, composed primarily of hydrocarbon compounds, provides protection against desiccation and other environmental challenges. Several of these cuticular hydrocarbon (CHC) compounds also function as semiochemical signals, and as such mediate pheromonal communications between members of the same species, or in some instances between different species, and influence behavior. Specialized cells referred to as oenocytes are regarded as the primary site for CHC synthesis. However, relatively little is known regarding the involvement of the oenocytes in the regulation of the biosynthetic, transport, and deposition pathways contributing to CHC output. Given the significant role that CHCs play in several aspects of insect biology, including chemical communication, desiccation resistance, and immunity, it is important to gain a greater understanding of the molecular and genetic regulation of CHC production within these specialized cells. The adult oenocytes of D. melanogaster are located within the abdominal integument, and are metamerically arrayed in ribbon-like clusters radiating along the inner cuticular surface of each abdominal segment. In this video article we demonstrate a dissection technique used for the preparation of oenocytes from adult D. melanogaster. Specifically, we provide a detailed step-by-step demonstration of (1) how to fillet prepare an adult Drosophila abdomen, (2) how to identify the oenocytes and discern them from other tissues, and (3) how to remove intact oenocyte clusters from the abdominal integument. A brief experimental illustration of how this preparation can be used to examine the expression of genes involved in hydrocarbon synthesis is included. The dissected preparation demonstrated herein will allow for the detailed molecular and genetic analysis of oenocyte function in the adult fruit fly.

Nestmate recognition and intraspecific chemical and genetic variation in Temnothorax ants

The evolutionary stability of social cooperation requires altruistic acts to be directed mainly towards related individuals. To maintain significant intragroup relatedness, social insects have evolved sophisticated recognition systems. We compared nestmate recognition abilities of three native European species of the ant genus Temnothorax in relation to variation in cuticular chemicals and intraspecific genetic diversity. Colony take-over and fusions are common in two sibling species T. nylanderi and T. crassispinus, but rare in T. unifasciatus, suggesting relatively inefficient nestmate recognition in the former species. Temnothorax nylanderi and T. crassispinus discriminated less effectively between nestmates and non-nestmates than T. unifasciatus. While chemical profiles of different colonies strongly overlapped in T. crassispinus and T. nylanderi, 97% of all T. unifasciatus samples were, in a discriminant analysis, correctly assigned to their respective colony. Chemical distances between colonies correlated with aggression levels in T. unifasciatus, but not in the other two species. Finally, preliminary genetic results indicated that haplotype divergence within T. nylanderi and T. crassispinus is remarkably low, whereas T. unifasciatus and other congeners are genetically more variable. In theory, loss of genetic variation, including depletion at loci involved in the production of cuticular hydrocarbons, could decrease colony recognition efficiency. We suggest that this might be the case in T. nylanderi and T. crassispinus, leading to colony fusions in the field. Despite their different life histories, these native species show similarities to invasive ants where genetic bottlenecks arising from founder effects are thought to result in a loss of colony boundaries.

Developmental isolation and subsequent adult behavior of Drosophila paulistorum. VI. Quantitative variation in cuticular hydrocarbons.

Our previous studies have demonstrated that long-term chemical contact with heterospecifics during development minimized unproductive heterospecific matings among the six Drosophila paulistorum semispecies. When socially isolated from conspecifics, discrimination significantly decreased so that more ultimately unproductive heterospecific matings occurred. Such results suggest that learning and social experiences play roles in mate recognition, using chemical information. In investigations into the development of discriminatory behavior in D. paulistorum, social experiences influence the production of cuticular hydrocarbons in both sexes among the six different semispecies of D. paulistorum. Produced by both sexes, 2-methyl triacontane has been detected from egg stages on, gradually increasing with age. However, 11-docosenyl acetate, a male pheromone, was not produced until early adult stages. There were consistent significant differences in quantities and ratios of each of these hydrocarbons among the six semispecies, contributing to their reproductive isolation. Furthermore, we found significant quantitative differences in hydrocarbons between socially isolated flies and communally raised ones: Socially isolated flies produced significantly greater quantities of hydrocarbons than communally raised flies, consistent with previous behavioral data, as isolated flies court more vigorously and often.

The genetics of mating recognition between Drosophila simulans and D. sechellia.

During courtship, visual and chemical signals are often exchanged between the sexes. The proper exchange of such signals ensures intraspecific recognition. We have examined the genetic basis of interspecific differences in male mating behaviour and pheromone concentration between Drosophila simulans and D. sechellia by using Drosophila simulans/D. sechellia introgression lines. Our results show a majority of quantitative trait loci (QTLs) explaining variation in both male mating behaviour and pheromone concentration to be located on the third chromosome. One QTL found on the third chromosome explains variation in time needed to start courtship and copulation as well as time spent courting. The position of such QTL (approximately 84A-88B) with effects on courtship and copulation aspects of mating includes the candidate sex determination gene doublesex (84E5-6) and Voila (86E1-2), a gene that affects male courtship in D. melanogaster. One additional third chromosome QTL explained variation in 7-tricosene pheromone concentrations among males. The interval mapping position of this QTL (approximately 68E-76E) did not overlap with the position detected for differences in mating behaviour and the intervals did not include candidate genes previously identified as having an effect on D. melanogaster cuticular hydrocarbon production. We did not detect any directionality of the effect of Drosophila sechellia allele introgressions in male mating recognition.

Effects of relative humidity, temperature, and population density on production of cuticular hydrocarbons in housefly Musca domestica L.

The production of cuticular hydrocarbons by both males and females of Musca domestica L. under very wet conditions (90% relative humidity) compared to the production at 50 and 20% relative humidity is delayed up to at least 3 days after emergence from the pupae. Eight days after emergence, however, males contain the same amounts of hydrocarbons at 90, 50, and 20% relative humidity, whereas females at 90% still possess less of these substances than at 50 and 20%. It is suggested that this is due to the fact that males, being more active than females, need more cuticular hydrocarbons to prevent water loss. No indication is found that relative humidity has a different effect on the production of the sex pheromone, muscalure [(Z)-9-tricosene] by females than on the production of the other hydrocarbons. Male and female flies produce more hydrocarbons at 35 degrees C than at 20 degrees C. On females, the relative amounts of nonacosane and methyl- and dimethylnonacosanes are significantly higher at 35 degrees C than at 20 degrees C. Female flies produce some (Z)-9-tricosene after eight generations at low population density in contrast to females at high population density, which did not produce muscalure. We suggest that because of the relatively large contribution to the total population, the properties of a small number of females are likely to be expressed sooner in the next generations of small populations than in those of large populations.

Control of female pheromones in Drosophila melanogaster by homeotic genes.

We have investigated the role of the Antennapedia and Bithorax complexes (ANT-C and BX-C) on the production of cuticular hydrocarbons in Drosophila melanogaster. In males, there is little, if any, influence of these complexes on the hydrocarbon pattern. In females, there are large and opposite effects of these complexes on diene production: two ANT-C mutations cause an increase in diene production and a reduction of monoenes, whereas most BX-C mutations result in a decrease in dienes and an increase in monoenes, although their sum remains constant. The effect is the highest in Mcp and iab6 females. It is suggested that a factor originating from the prothorax might activate the conversion of monoenes to dienes in females. The abdomen seems to have a crucial role in the production or control of pheromones: abdominal segments four to seven have the main effects, with a most dramatic effect for segments four and five.

Hormonal control of sex pheromone biosynthesis in Drosophila melanogaster

Production of cuticular hydrocarbons including the female sex pheromone cis,cis-7,11 heptacosadiene, was compared in intact and decapitated Drosophila melanogaster. Wild-type adults decapitated just after emergence showed very long-chain cuticular hydrocarbons (31–35 carbons) 3 days later, which are characteristic of young adults. Moreover among long-chain hydrocarbons (23–29 carbons) of decapitated females the sex pheromone level showed a marked decrease accompanied by an increase in monoenes. The quantitative effect strongly depended on the age of decapitation, being maximal when decapitation occurred within the first hour after emergence. Feeding had little influence on pheromone biosynthesis. A cephalic factor—yet to be characterized—seemed to exert the main control. Application of methoprene, a juvenile hormone (JH) analog, resulted in a decrease in very long-chain hydrocarbons in decapitated females but did not enhance the synthesis of sex pheromones. The analysis of mutants deficient in JH production confirmed the role of JH in the cuticular hydrocarbon switch.

Effects of allatectomy and ovariectomy on cuticular hydrocarbons in Calliphora vomitoria (diptera)

AbstractThe relationships between female attractiveness, cuticular hydrocarbons, and levels of juvenile hormone and ecdysteroids were studied in Calliphora vomitoria. The experiments were conducted at 48 and 72 h post‐emergence, according to attractiveness appearance and increase. The 48‐h‐old allatectomized females were less attractive than the control females, whereas no changes occurred either in cuticular hydrocarbons total mass production or in the different hydrocarbon families. However, the 72‐h‐old allatectomized females were more attractive than the control females, and, in relative proportions, allatectomy led to an increase in monomethylalkanes and a decrease in n‐alkanes.Only at 48 h were the ovariectomized females less attractive than the control females and did ovariectomy increase the relative proportions of monomethylalkanes. At 72 h, ovariectomy did not influence female attractiveness, but it decreased the total cuticular hydrocarbon production. Allatectomy and ovariectomy significantly decreased ecdysteroids levels at 48 and 72 h. Ovariectomy did not affect juvenile hormone production.These results suggest that attractiveness and cuticular hydrocarbon synthesis could be under the direct control of ecdysteroids and the indirect influence of juvenile hormone. © 1994 Wiley‐Liss, Inc.

Effets d'un élevage en isolement sur le développement gonadotrope, la production d'hydrocarbures cuticulaires et le comportement sexuel de Calliphora vomitoria (Diptères, Calliphoridae)

Isolated rearing (during the first gonadotropic cycle) was found to modify physiological variables (gonadotropic development, cuticular hydrocarbon production) as well as behavioral modalities of the courtship of male and female Calliphora vomitoria. In males, slower development and a decrease in the volume of the testes and the annex glands were noticed; in females, there was only a reduction in the volume of the terminal follicle without modification of the rate of growth. In contrast, cuticular hydrocarbon production was more disturbed in the female than in the male. Isolated rearing of males did not change either the number or the relative proportions and concentrations of cuticular hydrocarbons. In females, isolation was accompanied by a slight increase in both the relative proportions of cuticular hydrocarbons, namely monomethyalkanes, dimethylalkanes, and alkenes, and the concentrations of all hydrocarbons. Finally, isolated rearing was found to modify the sexual behavior of both partners. Isolated males showed later and scarcer sexual behavior than grouped males. They courted females later and more briefly, with lower frequencies and shorter durations of each stage of courtship; therefore, courtship was reduced with many interruptions. Isolated females appeared more permissive, with lower frequencies and durations of the stages of courtship, and shorter courtship. These differences were emphasized when both partners were reared in isolation. The lack of exchanges during imaginal development led, owing to lack of exercise and (or) nervous maturation, to sex-specific alterations: decreased gonadotropic development in males, modified cuticular hydrocarbon production in females, and reduced sexual behavior in both sexes.