Replenishment of Drosophila Male Pheromone After Mating

doi:10.21203/rs.3.rs-3568634/v1

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Replenishment of Drosophila Male Pheromone After Mating

https://doi.org/10.21203/rs.3.rs-3568634/v1

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published 25 Jan, 2024

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Insects exocrine gland products can be involved in sexual communication, defense, territory labelling, aggregation and alarm. In the vinegar fly Drosophila melanogaster the ejaculatory bulb synthesizes and releases 11-cis-Vaccenyl acetate (cVa). This pheromone, transferred to the female during copulation, affects aggregation, courtship and male-male aggressive behaviors. To determine the time-course of cVa replenishment, males of a control laboratory strain and from the desat1 pheromone-defective mutant strain were allowed to mate successively with several females. We measured mating frequency, duration and latency, the amount of cVa transferred to mated females and the residual cVa in tested males. Mating duration remained constant with multiple matings, but we found that the amount of cVa transferred to females declined with multiple matings, indicating that, over short, biologically-relevant periods, replenishment of the pheromone does not keep up with mating frequency, resulting in the transfer of varying quantities of cVa. Adult responses to cVa are affected by early developmental exposure to this pheromone; our revelation of quantitative variation in the amount of cVa transferred to females in the event of multiple matings by a male suggests variable responses to cVa shown by adults produced by such matings. This implies that the natural role of this compound may be richer than suggested by strictly-controlled laboratory experiments.

cis-Vaccenyl acetate

Copulation

Drosophila melanogaster

Cuticular hydrocarbon

Many insects possess exocrine glands involved in the synthesis and external release of semiochemicals used in inter- and intraspecific interactions (Noirot and Quennedey 1974; Ayasse et al. 2001; Aldrich and Zhang 2016; Costa-Leonardo et al. 2023). These chemicals can be involved in a variety of phenomena, such as defence, territory marking or mate recognition. Flies of the intensely studied Drosophila melanogaster species sub-group seem to possess a relatively simple system. They secrete cuticular hydrocarbons (CHCs) onto their epicuticle, probably through pore canals (Wang et al. 2020), some of which have a pheromonal role (Jallon 1984; Ferveur 2005); volatile compounds derived from the biosynthesis of these CHCs may also be involved in chemical communication (Lebreton et al. 2017). One of the most studied Drosophila pheromones is produced elsewhere – male flies synthesise 11-cis-vaccenyl acetate (cVa) in their ejaculatory bulb (EjB; Schaner et al. 1989a; Schaner et al. 1989b; Guiraudie-Capraz et al. 2007). This compound is released through the ejaculatory duct (EjD) and then transferred into the female during copulation (Butterworth 1969). It is involved in aggregation, egg-laying, pre-mating interactions, associative learning and male-male aggression (Jallon et al. 1981; Bartelt et al. 1985; Ejima et al. 2007; Wang et al. 2011; Duménil et al. 2016; Keesey et al. 2016). Other pheromone-releasing glands may exist, as hypothezised in both D. sechellia (Cobb and Ferveur 1996) and D. melanogaster (Lasbleiz et al. 2006) but their localization and the identity of any compound(s) are both unknown.

Shortly after mating, the high levels of cVa introduced into the female and released through her genital opening, act as an ‘anti-aphrodisiac’ on male flies, reducing her attractivity and the likelihood of a second mating, as a form of chemical mate guarding (Laturney and Billeter 2014; Brown et al., 2023). However, a few hours after insemination, some female Drosophila may eject the mating plug introduced by the male, consisting of most of the sperm, together with cVa and other compounds (Clark et al. 1995; Price et al. 1999; Laturney and Billeter 2016). Reducing the amount of cVa in the female, increases the probability of insemination by another male, perhaps providing the female with the opportunity of increasing the fitness of her offspring by mating with a higher quality male – in the case of a second mating, most of the sperm from the first mating is replaced (Snook and Hosken 2004). These behavioural changes are partly due to the physiological effects of a variety of male seminal proteins transferred by the male into the female reproductive tract (Billeter and Wolfner 2018). Developmental effects are also involved: the amount of cVa placed on eggs by the ovipositing female affects male flies derived from those eggs – both their courtship response to cVa and their ability to release this substance (Everaerts et al. 2018; Cortot et al. 2022b).

External compounds also affect post-mating behaviour in D. melanogaster. In particular, 7-tricosene (7-T), often found in high amounts on the male cuticle, is passively transferred onto the female during mating (Coyne et al. 1994; Everaerts et al. 2010). The post-mating combination of cVa and 7-T carried by the female strongly reduces her attractivity to males (Laturney and Billeter 2016). Most females carry 7,11-heptacosadiene (7,11-HD; Antony and Jallon, 1982): this pheromone stimulates conspecific male courtship and inhibits male courtship in the sibling D. simulans species (Jallon 1984; Savarit et al. 1999). It can be assumed that during mating, just as 7-T is passively transferred from the male to the female, 7,11-HD similarly moves from the female to the male. However, it is not known how such transfers affect the possibility of subsequent matings by either male or female.

While the behavioral effects induced by cVa have been carefully documented at subcellular and molecular levels (Benton et al. 2007; Kurtovic et al. 2007; Laughlin et al. 2008; Datta et al. 2008; Chertemps et al. 2012; Das et al. 2017), the rate at which the male replenishes his stock of cVa after mating remains unknown. This is significant, as the amount of cVa transferred to females during mating may affect both their behaviour and that of courting males. It cannot be assumed that the same amount of cVa is consistently transferred during mating, irrespective of the number of copulations by the male.

To explore the dynamics of post-copulatory replenishment in male Drosophila, we performed two experiments. First, over a two-hour period, single males were presented with single females and allowed to mate freely (Fig. 1A). Second, single males were tested on two successive days using a similar procedure (Fig. 1B). In females, we measured the amounts of transferred cVa and 7-T and the amount of 7,11-HD that remained on their cuticle; in males, measured their remaining amounts of cVa and 7-T and the amount of 7,11-HD found on their cuticle. All these measurements were then correlated with the frequency of mating and the interval between copulations. Additional insights were gained by comparing the Canton-S laboratory strain (Cs) and the desat1 mutant strain, which shows reduced cVa release (Cortot et al. 2022b) and very reduced 7-T and 7,11-HD biosynthesis (Marcillac et al. 2005a).

Flies. We used two Drosophila melanogaster stocks: the laboratory strain Canton-S (Cs) and the desat1¹⁵⁷³-Gal4 homozygous mutant strain (desat1) which produce a reduced quantity of 7-T and 7,11-HD (Marcillac et al. 2005a; Marcillac et al. 2005b); desat1 males also show reduced cVa emission (Cortot et al. 2022b). The desat1 stock was previously backcrossed with Cs over several generations to produce two strains with identical genetic background. Stocks were maintained on alcohol-free standard cornmeal medium mixed with killed yeast in 30 ml glass vials, at 24 ± 0.5°C and 65 ± 5% humidity on a 12:12 dark:light cycle. To avoid larval competition, only 8–10 pairs of progenitor flies were kept in each vial and transferred to fresh food vial every 2–3 days to generate tester flies. 1- to 2-hour old flies were sexed under light carbon dioxide anaesthesia 2–4 hours after lights on and were kept in fresh-food glass vials either in small groups of 5–7 flies for females or in isolation for males. All tested females were 4 days old while the age of tester males was either 2, 4 or 10 days old.

Behavior. Behavioral data were obtained between 20th March and 9th April 2020, using 579 males and 1057 females. For each experiment, one male was aspirated (without anaesthesia) under a watch glass, which formed a 1.6 cm³ observation chamber. After 10–20 minutes, a virgin female was introduced (this time was defined as T0). Each test was performed for 120 minutes in white light, at 24 ± 0.5°C. Each male was identified and his ability to move was checked – motionless males were discarded. We measured mating latency (the interval between T0 and copulation onset) with the first female (♀1) and copulation duration. Less than 3 minutes after the end of mating, ♀1 was aspirated into an empty glass vial kept on ice. A second female (♀2) was then introduced in the chamber with the male; the pair were allowed to mate and mating latency and duration were noted. After mating the female was put into a separate vial that was kept on ice ; each female was used only once. During the duration of this experiment (120 min), males were observed to copulate at most four times. Where males mated more than once, we measured the interval between consecutive copulations. At the end of the two-hour long test, each male was either individually transferred into an empty glass vial kept on ice (Experiment #1; Fig. 1A) or placed in a fresh food vial and retested the next day using the same procedure (Experiment #2; Fig. 1B). All empty vials containing tested male and female flies were rapidly transferred to -20° and the insects were subsequently subject to chemical extraction.

Chemical analysis. Frozen flies were individually immersed for 24 hours in 30µl dichloromethane at room temperature. This allowed us to extract the complete amount of cVa and CHCs present inside and outside of the fly (Cortot et al. 2022b). The solvent contained 3.33 ng/µl of C26 (n-hexacosane) and 3.33 ng/µl of C30 (n-triacontane) used as internal standards (ISs). Amounts of cVa and CHCs were quantified by gas chromatography using a Varian CP3380 gas chromatograph fitted with a flame ionization detector, a CP Sil 5CB column (25 m x 0.25 mm internal diameter; 0.1 µm film thickness; Agilent), and a split–splitless injector (60 ml/min split-flow; valve opening 30 sec after injection) with helium as the carrier gas (50 cm/sec at 120°C). The temperature program began at 120°C, ramping at 10°C/min to 140°C, then ramping at 2°C/min to 290°C, and holding for 10 min. The chemical identity of each peak was determined according to (Everaerts et al. 2010). The amount (ng/insect) of each compound was calculated based on the readings obtained from the ISs.

Statistics. We compared mating latency, mating duration and, in the case of multiple matings, mating interval for both Cs and desat1 pairs using the Kruskall-Wallis test with Monte-Carlo simulations, followed by Conover-Iman multiple pairwise comparisons (P = 0.05, with Bonferroni’s correction). The amounts of the various chemicals were compared using the same Kruskall-Wallis test, a Mann-Whitney test or a Wilkoxon signed rank test, where applicable. Mating frequencies were compared using the Wilks G² likelihood ratio test completed with a computation of significance by cell (Fisher's exact test). All statistical analyses were performed using XLSTAT Premium 2021.5.1.1220 (Addinsoft 2021).

Experiment #1. Male Mating With Multiple Females Within Two Hours.

Mating Behavior in Homotypic Matings. Table 1 summarises the mating statistics for Cs pairs. 178 males (73.3%) mated with at least one female; two males mated four times.

Table 1

*Courtship and mating performance in homotypic and heterotypic pairs.* We tested flies of two genotypes: control Cs (Cs) and *desat1* mutant flies. Values shown indicate the total number of pairs tested, courting and mating (N) or mating frequency (%) relative to the total number of males tested. These values are shown relative to the number of male matings over two hours.
		Courtship		Number of copulations
				0		1		2		3		4
Female x male	n	n	%	n	%	n	%	n	%	n	%	n	%
(Homotypic)
Cs x Cs	243	231	95.1	65	26.7	104	42.8	43	17.7	29	11.9	2	0.9
desat1 x desat1	231	212	91.8	151	65.4	74	32	6	2.6	0	0	0	0
(Heterotypic)
Cs x desat1	12	11	91.7	4	33.3	7	58.3	1	8.3	0	0	0	0
desat1 x Cs	10	9	90	3	30	3	30	2	20	1	10	1	10

We compared the performance of Cs males of different ages (2, 4 and 10-days old) and social experience (kept alone: x1; in groups of 5 : x5; x5 isolated one or two days before testing : -1S and − 2S; Suppl. Figure 1A). While the variation of these conditions did not alter the proportion or propensity of male mating, it slightly affected (i) mating latency (or the interval between successive mating) and (ii) copulation duration (Suppl. Figure 1B,C). More precisely, with ♀1 females 4-day old males reared alone showed a significantly shorter mating latency (23 ± 3.8 min) than 2-day old reared in similar conditions or 10 day old males held in groups of five (38.0 ± 4.8 min and 37.9 ± 4.8 min). Mating intervals did not vary between conditions. Mating duration was significantly shorter in 4-day old males held singly (17.1 ± 1.3 min) than in 10-day old males kept in groups (24.9 ± 0.9 min). There was no change in mating duration in males that mated more than once.

Table 1 summarises the mating statistics for desat1 males. 80 males (37.3%) mated with at least one female; six males mated twice. Age and experience significantly affected the proportion of males mating once (Suppl. Figure 2A): significantly more 4-day old males held singly mated (73.7%) than 10-day old males held in groups (20.0%) copulated compared to other males. Mating latency was significantly shorter in 2-day old males held singly (28.3 ± 5.6 min) than in 10-day old males held in the same conditions (53 ± 7.2 min; Suppl. Figure 2B). Mating durations with ♀1 were significantly shorter in 2 day-old males held singly (12.8 ± 0.4 min) than in 10 day-old males held singly or in 4 day-old males held in groups (16.1 ± 1.9 and 16.9 ± 0.8 min, respectively; Suppl. Figure 2C).

Overall, mating frequency was significantly lower in desat1 males than in Cs males, although courtship levels were similar (Fig. 2A). No difference was observed between the strains for mating latency or interval, although mating duration was consistently significantly longer in Cs than in desat1 males. Multiple component analysis of the behaviour of Cs and desat1 homotypic pairs revealed that mating latency and duration with ♀1 were positively correlated. Both parameters were negatively correlated with the number of male copulation(s) (Suppl. Figure 3A&B) – perhaps unsurprisingly, males who mated more quickly and with a shorter duration tended to have more matings in the 2-hour test period.

Mating Behavior in Heterotypic Matings. Although the sample sizes were relatively small, comparison of reciprocal pairs of the two genotypes (Cs female x desat1 male and desat1 female x Cs male) suggests that only the male genotype influenced the frequency of multiple male matings (Suppl. Figure 4). No differences between the two crosses were observed for mating latency with ♀1 flies while mating duration was significantly longer in desat1 x Cs than in Cs x desat1. Based on the differences between homotypic matings of these two strains, this indicates that mating duration depends on male genotype – see Fig. 2A.

Pheromones - cVa. We measured the amount of cVa transferred to females by males who mated with more than one female (Fig. 2B, left) and the amount of cVa left in those males (Fig. 2B, right). Pooling across male age and rearing conditions, Cs ♀1 flies received significantly more cVa than desat1 females (387 ± 11 and 273 ± 32 ng, respectively; P = 0.05). ♀2 flies received significantly less cVa (167 ± 30 ng; P = 0.05) with no significant differences between the two types of female. The amount of cVa received by ♀3 and ♀4 Cs individuals was not significantly different than in ♀2 flies. In males, residual cVa significantly decreased after the first and second matings (0 vs 1 and 1 vs 2; P = 0.05) in both strains – Cs males copulating once or twice retained 650 ± 64 and 404 ± 49 ng cVa respectively, while desat1 males copulating once or twice retained 483 ± 52 and 327 ± 59 ng cVa, respectively.

Pheromones − 7-T and 7,11-HD. Cuticular pheromones in both sexes showed only a slight variation depending on the number of matings by a male (Fig. 2C, D). The only significant variation was found for 7,11-HD, which increased between virgin and mated Cs males; no further significant increase was observed in males that mated more than once.

Experiment #2. Male Mating With Multiple Females On Two Successive Days

Mating Behavior. Cs males were relatively consistent in their mating performance over the two days of testing (D1, D2; Fig. 3A). Males that did not mate on D1 mated at most once on D2; males that mated once on D1 mated at most twice on D2. Males that mated twice on D1 mated up to 3 times on D2, while males that mated three times on D1 mated 2 or 3 times on D2 (3/2; 3/3). desat1 males were also relative consistent in their mating frequency on the two days (Fig. 3B).

Pheromones - cVa Transferred To Cs Females. Overall, Cs females that mated with virgin Cs males received an average of 337 ± 21 ng cVa, generally ranging between 270 and 400 ng cVa (in two outlying cases the figures were 154 and 525 ng). Females that mated with a male that had mated more than once received less cVa as the male mated more: ♀1 received significantly more cVa than ♀2 and ♀3 on both days (Fig. 4A; P < 0.001). The amounts of cVa received from males who had mated the same number of times showed a significant decrease on D2 for both in ♀1 and ♀2 individuals (P < 0.001 and P = 0.001, respectively), but not for ♀3 flies (Fig. 4B).

To evaluate the ability of males to replenish their cVa, we compared cVa levels between ♀1-D1 and ♀1-D2 flies that mated with males showing a similar D1 copulation performance (Fig. 4C). The cVa decrease was not significant in ♀1-D2 females mating with males which had copulated once on D1 but was highly significant with males that copulated twice on D1 (P < 0.001), suggesting that these males had not been able to sufficiently replenish their cVa levels overnight.

Pheromones - Residual cVa Left In Cs Males. The mean amount of cVa in an unmated male was 885 ng (D1 and D2 pooled; Fig. 3A). The levels of cVa in males that did not mate or mated once on D2 were not significantly different to this value, irrespective of their mating frequency on D1. Males that mated twice on D2 showed differences between them depending on their mating frequency on D1 – cVa levels were significantly lower in males that mated three times on D1 compared to males that mated once or twice (P = 0.028 and 0.019, respectively), suggesting that they had not sufficiently replenished their cVa levels. No differences were found for the few males that mated three times on D2. Differences were also observed between males that mated the same number of times on D1: males that mated once on D1 but twice on D2 showed significantly lower levels of cVa (compare 1/0 and 1/1 vs 1/2; P = 0.042); similar declines were observed between males that mates twice on D1 but did not mate on D2 (2/0) and those males that mated twice (2/2) or three times (2/3) on D2 (P = 0.019 and 0.034, respectively).

Pheromones - cVa Transferred To desat1 Females.

desat1 females that mated with virgin desat1 males received 120–440 ng cVa (mean = 337 ± 21 ng cVa). No significant variation was found between the two days, perhaps due to the small number of desat1 males mating (Fig. 5).

Pheromones - cVa Remaining In desat1 Males

Males with a similar number of matings on D2 did not vary in their cVa levels depending on the number of times they mated on D1 (Fig. 3B). Males with a similar number of D1 matings showed a significant cVa decrease according to their number of D2 matings (P < 0.001).

Pheromones − 7-T And 7,11-HD

No variation was observed in either female or male flies of either genotype, irrespective of the number of times the male had mated (Fig. 2C, D).

We measured two aspects of changes to pheromonal composition in Drosophila melanogaster males over time and with increasing numbers of matings. First, we measured the dynamics of the replenishment of cVa, a pheromone that is passed from males to females during mating (Fig. 2B); second, we measured the passive transfer of cuticular hydrocarbon pheromones. We performed these studies in two strains, a wildtype strain (Cs) and desat1, a mutant strain that shows altered levels of production and detection of cVa. These experiments provide a first insight into the fundamental physiological processes involved in Drosophila pheromone replenishment and their consequences for mating behaviour.

The amount of cVa transmitted to females significantly decreased when the males mated a second time within two hours, but showed no significant decline between third and fourth matings. Similarly, the residual cVa left in and on males significantly decreased between the first and second mating but showed no further decline on subsequent matings. This indicates that even if a male mates several times in a short space of time, he still retains a residual amount of cVa – about 25% of the initial amount. Over the short timescale of this experiment (two hours), there was no full replenishment of cVa; the constant level of cVa transferred and retained by males who mated more than twice suggests that there is a constant level of resynthesis and replenishment of the pheromone, which may be triggered by mating. Even twenty four hours was not sufficient to fully replenish a male’s level of cVa – males who mated twice on day one (D1) transferred significantly less cVa to their female partner when mating on day two (D2), compared to those males who mated only once on D1. However, if a male only mated once on D1, then the levels of cVa they transferred to a female on D2 were not significantly reduced (Fig. 3A; compare 0/0 vs 0/1; 1/0 vs 1/1 in Fig. 3).

These findings all apply to Cs flies. The results from the pheromonal mutant flies desat1 were different, partly because desat1 males showed reduced mating levels (Figs. 2A, 3B and 5). Unlike Cs flies, desat1 males who mated only once on D1 did not completely replenish their cVa levels after 24 hours. This may be related to their impaired ability to release cVa on their cuticle (Cortot et al. 2022a), due to misexpression of the desaturase1 gene in the male ejaculatory duct of desat1 males (Bousquet et al. 2012).

Most studies of insect exocrine gland replenishment have focused on the release of defensive chemicals (Blum 1981; Baldwin et al. 1990). When attacked, the kissing bug Triatoma infestans, a trypanosome vector, discharges large amounts of its secretion but retains around 25% which can be subsequently discharged (Palottini et al. 2014). The viviparous Pacific beetle cockroach, Diploptera punctata, retains 11% of the initial content even after several stimulations (Baldwin et al. 1990). These figures suggest that there is a short-term constant replenishment of the gland, but that this is not sufficient to return the insect to its pre-stimulation level. In other insects, the full replenishment rate varies between two weeks (in the prothoracic glands of the American water beetle species Agabus seriatus and A. obtusatus; (Fescemyer and Mumma 1983) to several months (in D. punctata tracheal glands; Baldwin et al. 1990). These dynamics do not seem to be correlated with the efficiency of the biosynthetic pathway of defensive chemicals (Blum 1987; Laurent et al. 2005), but instead to be affected by the developmental stage and by food availability – some insects can sequester plant compounds for use in their defense glands (Bowers, 1990; (Whitman et al. 1992; Palottini et al. 2014). Chemical replenishment in exocrine glands can vary depending on species, sex and developmental stage (Kearsley and Whitham 1992; Rossini et al. 1997; Camara 1997; Yu et al. 2023).

The passive transfer of two major sex-specific cuticular pheromones (male 7-T and female 7,11-HD) during mating revealed that, as expected, males receive female-specific 7,11-HD (experiments involving large groups of flies resulted in the overnight passive transfer of substantial CHC amounts from donor to receiver flies (Coyne et al. 1994; Savarit et al. 1999; Grillet et al. 2006). However, we found no significant 7,11-HD increase in the amount detected with increasing numbers of mating. No significant change in either pheromone was detected in females. No significant effect was detected in desat1 flies probably because of their low initial 7-T and 7,11-HD levels. Previously, Cs males that mated only once were shown to have a significant decrease in their levels of 7-T and the appearance of low levels of 7,11-HD, two days after mating (Everaerts et al. 2010). Here we found that following mating, males showed no significant variation of 7-T. This discrepancy may be due to differences in the resolution of the analytical methods used in the two studies.

Male mating performance differed between Cs and desat1 homotypic pairs, due to the higher mating frequency of Cs males (Fig. 2A; Suppl. Figures 1 and 2). Cs males also copulated faster and showed a longer copulation duration than desat1 males. Young males of both strains who were kept alone mated more quickly and for a shorter time compared to older flies and flies kept in groups (Suppl. Figure 3). Males tended to show the same number of matings when they were tested on two subsequent days (Fig. 3) indicating that physiological and or behavioural factors underlying successful mating were constant in individuals, perhaps revealing inter-individual variability.

Responses to cVa are altered by early experience – the pheromone is deposited on the egg and in the food by the female, and larval contact alters subsequent adult behaviours involving cVa such as food finding, mate finding and male courtship (Everaerts et al. 2018; (Tolassy et al. 2023). We have shown that the amount of cVa transferred to females declines where males have previously mated; this suggests that the amount of cVa available to produce such developmental conditioning effects may vary between females and constitute a significant source of behavioural variation in their offspring. The physiological and evolutionary consequences of polyandry and female remating have been studied much more intensively than the consequence of polygyny (Fuerst et al. 1973; Imhof et al. 1998; Laturney et al. 2018). Our study shows that polygyny not only involves the insemination of multiple females, it can also lead to changing pheromonal effects on both the female and potentially on her offspring, due to declining levels of pheromone in the male. We believe that the present study will provide new insights into the understanding of insect chemical communication and reproduction and also suggests how more naturalistic experimental designs, involving multiple matings in a short space of time, may reveal effects that occur not only in the laboratory but also in nature, shaping the evolution of wild populations.

Author Contributions Statement

J-F.F. performed the experiments J-F.F and C.E. analyzed the data. J_F.F, M.C., B.M. and C.E. wrote the ms text. C.E. prepared the figures. All authors reviewed the manuscript.

Competing Interests

The authors declare that they have no competing interests as defined by Springer, or other interests that might be perceived to influence the results and/or discussion reported in this paper.

Funding: This work was funded by the University of Bourgogne-Franche Comté, the Bourgogne-Franche Comté Regional Council (PARI 2016), the Centre National de la Recherche Scientifique (CNRS; Insb) and Campus France (PHC Procope 49359TF grant).

Addinsoft (2021) XLSTAT 2021: Data analysis and statistical solution for Microsoft Excel. Addinsoft, Paris, France
Aldrich JR, Zhang Q-H (2016) Chemical ecology of Neuroptera. Annu Rev Entomol 61:197–218. 10.1146/annurev-ento-010715-023507
Ayasse M, Paxton RJ, Tengo J (2001) Mating behavior and chemical communication in the order Hymenoptera. Annu Rev Entomol 46:31–78. 10.1146/annurev.ento.46.1.31
Baldwin IT, Dusenbery DB, Eisner T (1990) Squirting and refilling: dynamics of p-benzoquinone production in defensive glands of Diploptera punctata. J Chem Ecol 16:2823–2834. 10.1007/BF00979476
Bartelt RJ, Schaner AM, Jackson LL (1985) cis-Vaccenyl acetate as an aggregation pheromone in Drosophila melanogaster. J Chem Ecol 11: 1747–1756 10.1007/bf01012124
Benton R, Vannice KS, Vosshall LB (2007) An essential role for a CD36-related receptor in pheromone detection in Drosophila. Nature 450:289–U213. 10.1038/nature06328
Billeter J-C, Wolfner MF (2018) Chemical cues that guide female reproduction in Drosophila melanogaster. J Chem Ecol 44:750–769 doi:0.1007/s10886-018-0947-z
Blum MS (1981) Chemical defenses of Arthropods. Academic Press Inc, New York
Blum MS (1987) Biosynthesis of Arthropod exocrine compounds. Annu Rev Entomol 32:381–413. 10.1146/annurev.en.32.010187.002121
Bousquet F, Chauvel I, Flaven-Pouchon J, Farine J-P, Ferveur J-F (2012) Expression of a desaturase gene, desat1, in neural and nonneural tissues separately affects perception and emission of sex pheromones in Drosophila. Proc Natl Acad Sci U S A 109:249–254 10.1073/pnas.1109166108
Butterworth FM (1969) Lipids of Drosophila: a newly detected lipid in the male. Science 163:1356–1357. 10.1126/science.163.3873.1356
Camara MD (1997) Physiological mechanisms underlying the costs of chemical defence in Junonia coenia Hübner (Nymphalidae): a gravimetric and quantitative genetic analysis. Evol Ecol 11:451–469. 10.1023/A:1018436908073
Chertemps T, Francois A, Durand N, Rosell G, Dekker T, Lucas P, Maibeche-Coisne M (2012) A carboxylesterase, Esterase-6, modulates sensory physiological and behavioral response dynamics to pheromone in Drosophila. BMC Biol 10:56. 10.1186/1741-7007-10-56
Clark AG, Aguade M, Prout T, Harshman LG, Langley CH (1995) Variation in sperm displacement and its association with accessory-gland protein loci in Drosophila melanogaster. Genetics 139:189–201. 10.1093/genetics/139.1.189
Cobb M, Ferveur J-F (1996) Evolution and genetic control of mate recognition and stimulation in. Drosophila Behav Process 35:35–54. 10.1016/0376-6357(95)00052-6
Cortot J, Farine J-P, Cobb M, Everaerts C, Ferveur J-F (2022a) Factors affecting the biosynthesis and emission of a Drosophila pheromone. J Exp Biol 225. 10.1242/jeb.244422
Cortot J, Farine J-P, Ferveur J-F, Everaerts C (2022b) Aging-related variation of cuticular hydrocarbons in wild type and variant Drosophila melanogaster. J Chem Ecol 48:152–164. 10.1007/s10886-021-01344-0
Costa-Leonardo AM, da Silva IB, Laranjo LT (2023) Termite exocrine systems: a review of current knowledge. Entomol Exp Appl 171:325–342. 10.1111/eea.13292
Coyne JA, Crittenden AP, Mah K (1994) Genetics of a pheromonal difference contributing to reproductive isolation in Drosophila. Science 265:1461–1464
Das S, Trona F, Khallaf MA, Schuh E, Knaden M, Hansson BS, Sachse S (2017) Electrical synapses mediate synergism between pheromone and food odors in Drosophila melanogaster. Proc Natl Acad Sci U S A 114:E9962–E9971. 10.1073/pnas.1712706114
Datta SR, Vasconcelos ML, Ruta V, Luo S, Wong A, Demir E, Flores J, Balonze K, Dickson BJ, Axel R (2008) The Drosophila pheromone cVA activates a sexually dimorphic neural circuit. Nature 452:473–477. 10.1038/nature06808
Duménil C, Woud D, Pinto F, Alkema JT, Jansen I, Van Der Geest AM, Roessingh S, Billeter J-C (2016) Pheromonal cues deposited by mated females convey social information about egg-laying sites in Drosophila melanogaster. J Chem Ecol 42:259–269. 10.1007/s10886-016-0681-3
Ejima A, Smith BPC, Lucas C, Van Naters WV, Miller CJ, Carlson JR, Levine JD, Griffith LC (2007) Generalization of courtship learning in Drosophila is mediated by cis-vaccenyl acetate. Curr Biol 17:599–605
Everaerts C, Cazalé-Debat L, Louis A, Pereira E, Farine J-P, Cobb M, Ferveur J-F (2018) Pre-imaginal conditioning alters adult sex pheromone response in Drosophila. PeerJ eCollection 2018. 10.7717/peerj.5585
Everaerts C, Farine J-P, Cobb M, Ferveur J-F (2010) Drosophila cuticular hydrocarbons revisited: mating status alters cuticular profiles. PLoS ONE 5:e9607. 10.1371/journal.pone.0009607
Ferveur J-F (2005) Cuticular hydrocarbons: their evolution and roles in Drosophila pheromonal communication. Behav Genet 35:279–295. 10.1007/s10519-005-3220-5
Fescemyer HW, Mumma RO (1983) Regeneration and biosynthesis of dytiscid defensive agents (Coleoptera: Dytiscidae). J Chem Ecol 9:1449–1464. 10.1007/BF00988511
Fuerst PA, Pendlebury WW, Kidwell JF (1973) Propensity for multiple mating in Drosophila melanogaster females. Evolution 27:265–268. 10.1111/j.1558-5646.1973.tb00671.x
Grillet M, Dartevelle L, Ferveur J-F (2006) A Drosophila male pheromone affects female sexual receptivity. Proc R Soc Biol Sci Ser B 273:315–323. 10.1098/rspb.2005.3332
Guiraudie-Capraz G, Pho DB, Jallon JM (2007) Role of the ejaculatory bulb in biosynthesis of the male pheromone cis-vaccenyl acetate in Drosophila melanogaster. Integr Zool 2:89–99. 10.1111/j.1749-4877.2007.00047.x
Imhof M, Harr B, Brem G, Schlötterer C (1998) Multiple mating in wild Drosophila melanogaster revisited by microsatellite analysis. Mol Ecol 7:915–917. 10.1046/j.1365-294x.1998.00382.x
Jallon JM (1984) A few chemical words exchanged by Drosophila during courtship and mating. Behav Genet 14:441–478. 10.1007/BF01065444
Jallon JM, Antony C, Benamar O (1981) An anti-aphrodisiac produced by Drosophila melanogaster males and transferred to females during copulation. C R Acad Sci Ser III Sci Vie 292:1147–1149
Kearsley MJC, Whitham TG (1992) Guns and butter: a no cost defense against predation for Chrysomela confluens. Oecologia 92:556–562. 10.1007/BF00317849
Keesey IW, Koerte S, Retzke T, Haverkamp A, Hansson BS, Knaden M (2016) Adult frass provides a pheromone signature for Drosophila feeding and aggregation. J Chem Ecol 42:739–747. 10.1007/s10886-016-0737-4
Kurtovic A, Widmer A, Dickson BJ (2007) A single class of olfactory neurons mediates behavioural responses to a Drosophila sex pheromone. Nature 446:542–546. 10.1038/nature05672
Lasbleiz C, Ferveur J-F, Everaerts C (2006) Courtship behaviour of Drosophila melanogaster revisited. Anim Behav 72:1001–1012. 10.1016/j.anbehav.2006.01.027
Laturney M, Billeter J-C (2014) Neurogenetics of female reproductive behaviors in Drosophila melanogaster. Adv Genet 85:1–108. 10.1016/B978-0-12-800271-1.00001-9
Laturney M, Billeter J-C (2016) Drosophila melanogaster females restore their attractiveness after mating by removing male anti-aphrodisiac pheromones. Nat Communication 7:12322. 10.1038/ncomms12322
Laturney M, van Eijk R, Billeter J-C (2018) Last male sperm precedence is modulated by female remating rate in Drosophila melanogaster. Evol Lett 2:180–189. 10.1002/evl3.50
Laughlin JD, Ha TS, Jones DNM, Smith DP (2008) Activation of pheromone-sensitive neurons is mediated by conformational activation of pheromone-binding protein. Cell 133:1255–1265. 10.1016/j.cell.2008.04.046
Laurent P, Braekman J-C, Daloze D (2005) Insect chemical defense. In: Schulz S (ed) The chemistry of pheromones and other semiochemicals II. Springer Berlin Heidelberg, Berlin, Heidelberg, pp 167–229. doi:10.1007/b98317
Lebreton S, Borrero-Echeverry F, Gonzalez F, Solum M, Wallin EA, Hedenstroem E, Hansson BS, Gustavsson AL, Bengtsson M, Birgersson G, Walker WB III, Dweck JKM, Becher PG, Witzgall P (2017) A Drosophila female pheromone elicits species-specific long-range attraction via an olfactory channel with dual specificity for sex and food. BMC Biol 15. 10.1186/s12915-017-0427-x
Marcillac F, Bousquet F, Alabouvette J, Savarit F, Ferveur J-F (2005a) A mutation with major effects on Drosophila melanogaster sex pheromones. Genetics 171:1617–1628. 10.1534/genetics.104.033159
Marcillac F, Grosjean Y, Ferveur J-F (2005b) A single mutation alters production and discrimination of Drosophila sex pheromones. Proc R Soc Lon B 272:303–309. 10.1098/rspb.2004.2971
Noirot C, Quennedey A (1974) Fine structure of insect epidermal glands. Annu Rev Entomol 19:61–80. 10.1146/annurev.en.19.010174.000425
Palottini F, González A, Manrique G (2014) Filling dynamics of the Brindley’s glands in the blood-sucking bug Triatoma infestans (Hemiptera: Reduviidae). J Insect Physiol 71:122–127. 10.1016/j.jinsphys.2014.10.012
Price CSC, Dyer KA, Coyne JA (1999) Sperm competition between Drosophila males involves both displacement and incapacitation. Nature 400:449–452. 10.1038/22755
Rossini C, Attygalle AB, González A, Smedley SR, Eisner M, Meinwald J, Eisner T (1997) Defensive production of formic acid (80%) by a carabid beetle (Galerita lecontei). Proc Natl Acad Sci U S A 94:6792–6797. 10.1073/pnas.94.13.6792
Savarit F, Sureau G, Cobb M, Ferveur J-F (1999) Genetic elimination of known pheromones reveals the fundamental chemical bases of mating and isolation in Drosophila. Proc Natl Acad Sci U S A 96:9015–9020
Schaner AM, Benner AM, Leu RD, Jackson LL (1989a) Aggregation pheromone of Drosophila mauritiana, Drosophila yakuba and Drosophila rajasekari. J Chem Ecol 15:1249–1257. 10.1007/BF01014827
Schaner AM, Graham KJ, Jackson LL (1989b) Aggregation pheromone characterization and comparison in Drosophila ananassae and Drosophila bipectinata. J Chem Ecol 15:1045–1055. 10.1007/BF01015198
Snook RR, Hosken DJ (2004) Sperm death and dumping in Drosophila. Nature 428:939–941. 10.1038/nature02455
Tolassy V, Cazalé-Debat L, Houot B, Reynaud R, Heydel J-M, Ferveur J-F, Everaerts C (2023) Drosophila free-flight odor tracking is altered in a sex-specific manner by preimaginal sensory exposure. J Chem Ecol 49:179–194. 10.1007/s10886-023-01416-3
Wang L, Han X, Mehren J, Hiroi M, Billeter JC, Miyamoto T, Amrein H, Levine JD, Anderson DJ (2011) Hierarchical chemosensory regulation of male-male social interactions in Drosophila. Nat Neurosci 14:757–762. 10.1038/nn.2800
Wang Y, Norum M, Oehl L, Yang Y, Zuber R, Yang J, Farine J-P, Gehring N, Flötenmeyer M, Ferveur J-F, Moussian B (2020) Dysfunction of Oskyddad causes Harlequin-type ichthyosis-like defects in Drosophila melanogaster. PLoS Genet 16:e1008363. 10.1371/journal.pgen.1008363
Whitman DW, Jones CG, Blum MS (1992) Defensive secretion production in lubber grasshoppers (Orthoptera: Romaleidae): influence of age, sex, diet, and discharge frequency. Ann Entomol Soc Am 85:96–102. 10.1093/aesa/85.1.96
Yu P, Iwanami T, Yazaki H, Tsubuki M, Saito K, Hayashi F (2023) Cost of defensive spraying by larval Osmylus hyalinatus (Neuroptera: Osmylidae) for post-larval development. J Ethol 41:129–139. 10.1007/s10164-023-00779-0

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Replenishment of Drosophila Male Pheromone After Mating

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Abstract

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INTRODUCTION

METHODS AND MATERIALS

RESULTS

Experiment #2. Male Mating With Multiple Females On Two Successive Days

Pheromones - cVa Remaining In desat1 Males

Pheromones − 7-T And 7,11-HD

Discussion

Declarations

References

Additional Declarations

Supplementary Files

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