Sexual plasticity, the ability to modify sexual state or reproductive behavior in response to changing social conditions, is observed in both vertebrates and invertebrates (Bruce, 1959; Koene and Ter Maat, 2007; Roberts et al., 2012; Steiger et al., 2008; Yagound et al., 2012). In rodents, exposure to unfamiliar males often leads to the sudden termination of pregnancy, known as the Bruce effect. It is induced by male urinary peptides, such as MHC I peptides, activating the vomeronasal organ (Becker and Hurst, 2008; Leinders-Zufall et al., 2004; Zipple et al., 2019). This effect enhances reproductive fitness of both sexes, by eliminating the offspring of competing males and enabling females to select better mates even after conception. Many species also adapt their reproductive behavior in response to the social-sexual context change (SSCC), involving encounters with new sexual partners or competitors. Understanding the neural circuit mechanisms behind female responses to SSCC emerges as a central focus of neuroscience (Gaspar et al., 2022; Kim et al., 2019; Liu et al., 2022; Wei et al., 2021).

Drosophila melanogaster, the fruit fly, displays various social behaviors like aggregation, aggression, and sexual behavior (Bartelt et al., 1985; Billeter et al., 2006; Billeter and Levine, 2015). Similar to rodents, they primarily use the olfactory system to communicate socially through pheromones (Kohl et al., 2015; Sengupta and Smith, 2014). Some of these pheromones act as aphrodisiacs, while others regulate aggression or foster aggregation. For instance, 11-cis-vaccenyl acetate (cVA) attracts females but repels males and promotes aggregation (Billeter and Levine, 2015; Kurtovic et al., 2007; Mane et al., 1983). 7-Tricosene (7-T), a cuticular hydrocarbon (CHC) present in males, is an aphrodisiac to females and affects social interactions between males (Grillet et al., 2006; Wang et al., 2011). On the other hand, 7,11-heptacosadiene (7,11-HD), a related female-specific pheromone, functions as an aphrodisiac to males, triggering courtship behavior and involving species recognition (Antony et al., 1985; Toda et al., 2012).

The fruit fly’s chemosensory organs, located in various parts of the body, detect these pheromones (Ali et al., 2022; Joseph and Carlson, 2015). Olfactory receptor neurons (ORNs) in the sensilla of the antennae and maxillary palp are responsible for the detection of long-range volatile pheromones like cVA, while short-range pheromones like 7-T are sensed by neurons on the forelegs and labellum (Joseph and Carlson, 2015; Kohl et al., 2015; Sengupta and Smith, 2014).

The olfactory receptor Or47b, expressed in ORNs located in the at4 trichoid sensilla on the third antennal segment, is involved in several sociosexual interactions, including male mating success, mate preference, and female aggression toward mating pairs (Gaspar et al., 2022; Kohlmeier et al., 2021; Lone et al., 2015; Lone and Sharma, 2012; Zhuang et al., 2016). In males, Or47b senses fatty acid methyl esters and fatty acids that affect mating competition and copulation (Dweck et al., 2015; Lin et al., 2016). While the role of Or47b in female aggression is well established (Gaspar et al., 2022), its involvement in female sexual behavior is uncertain. In both sexes, Or47b ORNs project to VA1v glomeruli, where VA1v projection neurons receive their signal and project to the mushroom body calyx and lateral horn. Male Or47b neurons connect to neurons such as aSP5, aSP8, and aSP9, which express a male-specific transcription factor Fru^M (Yu et al., 2010).

CHC pheromones, which function as short-range pheromones, are detected primarily by neurons on the forelegs and the labellum that express gustatory receptors, ionotropic receptors, or the ppk/DEG-ENaC family of sodium channels (Joseph and Carlson, 2015; Kohl et al., 2015; Sengupta and Smith, 2014). CHCs like 7-T and 7,11-HD are sensed by ppk23-expressing M and F cells in the tarsi (Liu et al., 2020). 7-T and cVA are sensed by M cells expressing ppk23, whereas 7,11-HD and 7,11-nonacosadiene are sensed by F cells expressing ppk23, ppk25, and ppk29. In males, 7-T or 7,11-HD affects the neuronal activity of the Fru^M-expressing P1 neurons (Inagaki et al., 2014; Kohatsu et al., 2011; Sato and Yamamoto, 2020). However, how these CHCs signal in the female brain remains unknown.

Sperm ejection is a process by which females can remove the male ejaculate or the mating plug after copulation. This phenomenon has been observed in several animal species including feral fowl (Pizzari and Birkhead, 2000), black-legged kittiwake (Wagner et al., 2004), and dunnock (Davies, 1983). In the fruit fly, it typically occurs approximately 90 min after mating (Lee et al., 2015). This specific interval, referred to as ejaculate holding period (EHP), is thought to affect sperm usage and fecundity (Lee et al., 2015; Manier et al., 2010). The neurosecretory neurons in the brain pars intercerebralis (PI) that produce diuretic hormone 44 (Dh44), an insect orthologue of the corticotropin-releasing factor, regulate EHP (Lee et al., 2015). There is evidence that Drosophila females sense the social-sexual context through sperm ejected by other females. For instance, females were likely to lay more eggs when placed on a food patch containing male ejaculate deposited by other females (Duménil et al., 2016). However, it remains unknown whether the SSCC influences sperm ejection and EHP.

Female pC1 neurons, which express a specific transcription factor Dsx^F, integrate olfactory and auditory cues associated with male courtship (Lee et al., 2002; Zhou et al., 2014). The pC1 neurons, their male counterparts (i.e. P1 neurons), and the ventrolateral subdivision of ventromedial hypothalamus (VMHvl) neurons in mice share conserved circuit configurations and demonstrate functional similarity in coordinating social and sexual behaviors (Anderson, 2016; Jiang and Pan, 2022). There are 14 Dsx-positive pC1 neurons in each hemisphere of the brain, responsive to the male sex-pheromone cVA and courtship songs (Kim et al., 2024; Zhou et al., 2014). Connectome analyses identified 10 pC1 neurons that fall into five subtypes, with pC1a, b, and c subtypes associated with mating behavior and pC1d and e subtypes associated with aggression (Chiu et al., 2023; Deutsch et al., 2020; Han et al., 2022; Kim et al., 2024; Schretter et al., 2020; Wang et al., 2020). Although direct evidence connecting pC1 neurons to sperm ejection is limited, they are promising candidates for regulating sperm ejection or EHP, because sperm ejection allows females to eliminate the mating plug and male ejaculate, thereby restoring sexual attractiveness (Laturney and Billeter, 2016).

In this study, we demonstrated that two male pheromones, 2-methyltetracosane (2MC) and 7-T, significantly reduced the EHP through Or47b neurons and ppk23 neurons, respectively. These pheromone pathways converge on pC1 neurons, where they increase cAMP levels. The elevated cAMP in pC1 neurons resulted in a reduction of the EHP to a degree that was comparable to the effects of the male pheromones. It also enhanced the excitability of pC1 neurons, making them more responsive to both olfactory and auditory male courtship cues and promoting further mating following the earlier removal of the mating plug. These findings establish a novel behavioral paradigm that sheds light on the intricate molecular and neuronal pathways underlying female sexual plasticity.

Males employ a diverse range of strategies to enhance their reproductive fitness. One such strategy involves the formation of a ‘mating plug’, a mechanism that prevents females from engaging in further mating, thereby increasing fertilization success rates (Dixson, 1998; Parker, 1970; Schneider et al., 2016). As a means of intra-sexual competition, rival males often promote the removal or precocious expulsion of the mating plug. The evolution of this strategy is driven by intersexual interactions with polyandrous females, who often remove the mating plug to engage in additional mating with males of superior traits or higher social status than their previous partners (Dean et al., 2011; Pizzari and Birkhead, 2000). In the dunnock Prunella modularis, a small European passerine bird, the male often engages in cloacal pecking of mated females, inducing the expulsion of the previous mate’s sperm and mating plug, thereby increasing their chances of successful mating (Davies, 1983). In this study, we discovered that in D. melanogaster, freshly mated females exhibit an earlier removal of the mating plugs or a shorter EHP when kept with actively courting males. This behavior is primarily induced by the stimulation of females via male sex pheromones. In addition, our study has revealed that the neural circuit that processes male courtship cues and controls mating decisions plays an important role in regulating this behavior. This fly circuit has recently been proposed to be homologous to VMHvl in the mouse brain (Anderson, 2016; Jiang and Pan, 2022). By delving into the molecular and neuronal mechanisms underlying MIES, our study provides valuable insights into the broader aspect of behaviors induced by changes in the social-sexual context.

Our findings highlight the involvement of the Or47b receptor and Or47b ORNs in MIES. These OR and ORNs have been implicated in a range of social and sexual behaviors in both male and female fruit flies (Dweck et al., 2015; Gaspar et al., 2022; Kohlmeier et al., 2021; Lin et al., 2016; Lone et al., 2015; Lone and Sharma, 2012; Zhuang et al., 2016). Methyl laurate and trans-palmitoleic acid are odorant ligands for Or47b that account for many of these functions, particularly in males (Dweck et al., 2015; Lin et al., 2016). In this study, we provide compelling evidence that 2MC induces cAMP elevation in pC1 neurons and EHP shortening via both the Or47b receptor and Or47b ORNs, suggesting that 2MC functions as an odorant ligand for Or47b. Notably, gas chromatography-mass spectrometry analysis of CHCs from 4-day-old wild-type D. melanogaster revealed the presence of 2MC exclusively in males (Dweck et al., 2015). Surprisingly, however, unlike 2MC, neither methyl laurate nor trans-palmitoleic acid affected EHP. The reason for this paradoxical result remains unclear. A plausible interpretation is that the EHP shortening induced by 2MC may require not only Or47b but also other as yet unidentified ORs. With the establishment of a behavioral and cellular assessment of 2MC activity, the search for additional odorant receptors responsive to 2MC is now feasible. Another important avenue for further research is whether 2MC can also elicit behaviors previously associated with methyl laurate or trans-palmitoleic acid, such as promoting male copulation and courtship (Dweck et al., 2015; Lin et al., 2016).

We observed that both 2MC and 7-T exhibit both cellular and behavioral activity within a specific concentration range (Figure 3—figure supplement 3, Figure 4—figure supplement 1, Figure 5—figure supplement 2). This observation is of particular interest, given the multitude of environmental and biological factors that influence the levels of 2MC and 7-T, potentially affecting the capacity of males to induce MIES. For instance, exposure to low temperatures during development has been linked to increased production of both 2MC and 7-T (Bontonou et al., 2013). Similarly, mutation of the desiccation stress gene CG9186, which encodes a protein associated with lipid droplets, has been found to impact 2MC levels (Werthebach et al., 2019). Furthermore, 2MC levels rise with age in males (Everaerts et al., 2010). Thus, we propose that levels of 2MC, and possibly 7-T, may serve as indicators of male age and resilience to environmental stress in a complex manner.

In mated females, treatment with 2MC or 7-T increases cAMP levels in pC1b, c neurons but not in pC1a neurons. In contrast, pC1a neurons in virgin females are fully responsive to both male pheromones, showing increases in cAMP levels that are similar to those of pC1b,c neurons (Figure 5—figure supplement 3). The absence of cAMP levels in pC1a neurons in mated females likely results from the mating signal (i.e. sex peptide) silencing pC1a neurons. Connectome and electrophysiology data support this interpretation, as SAG neurons, which relay sex peptide signals, have the strongest synaptic connection with the pC1a among five pC1 subtypes (Wang et al., 2020). However, the activity of SAG neurons may also influence pC1c neurons, as they also have substantial synaptic connections with pC1c neurons, as seen in the hemibrain connectome dataset (Scheffer et al., 2020). Future studies are needed to understand the role of SAG neurons in the regulation of EHP.

We found that increased cAMP levels cause pC1b, c neurons in mated females, which are typically unresponsive to male courtship cues like cVA and pulse song, to become responsive and exhibit strong Ca²⁺ transients. Since pC1b, c neurons play a role in generating sexual drive and increasing female receptivity to male courtship, the 2MC- or 7-T-induced increases in cAMP are likely to control the removal of the mating plug and the engagement of mated females in further mating (Figure 7). This hypothesis aligns well with the previous report that mating reduces the sensitivity of Or47b ORNs, which we found to be responsive to 2MC, leading to an increased preference for pheromone-rich males after mating (Kohlmeier et al., 2021). Moreover, the finding that 2MC and 7-T induce cAMP levels in pC1b, c neurons in virgin females suggests that virgin females may also use 2MC and 7-T as odorant cues to assess male quality during their first mating. Indeed, females seem to evaluate male quality by the amount of 7-T, as increased 7-T promotes mating receptivity and shortens mating latency (Grillet et al., 2006).

Physiological factors like the nutritional status of females prior to mating and the nutritional status of their mates have been shown to influence EHP (Kim et al., 2023), and therefore potentially MIES. Hence, it is highly probable that MIES is regulated by additional central neurons such as Dh44-PI neurons that regulate these processes (Lee et al., 2015). However, it remains unclear whether and how Dh44-PI neurons and pC1 neurons interact to modulate EHP and MIES. The observation that double knockdown of Dh44R1 and Dh44R2 has only a marginal effect on MIES suggests that Dh44-PI neurons may also function independently of pC1 neurons, raising the possibility that multiple independent central circuits may contribute to the production of MIES.

Our initial screening of ORNs responsible for MIES revealed the involvement of Or47b ORNs, as well as several other ORNs. In addition to 2MC, which acts through Or47b-expressing ORNs, our findings indicate that 7-T and ppk23 neurons, which are capable of detecting 7-T, also play a role in MIES induction. In D. melanogaster and other related species, food odors typically serve as volatile long-range signals that attract both males and females (Lin et al., 2015; Verschut et al., 2023), suggesting that specific food odors may also influence EHP (Duménil et al., 2016). The involvement of multiple ORNs in the regulation of EHP predicts that pC1 neurons may process multiple odorants, not limited to those associated with mating behavior, including food odors. Future studies will explore the full spectrum of odorants processed by pC1 neurons in the regulation of EHP.

In conclusion, we have identified a circuit that, via the detection of a novel male pheromone, potentially signals male quality and governs the female’s decision to remove the mating plug of her last mate and mate again.

All data generated or analysed during this study are included in the manuscript and supporting files.

1. 1. Ali MZ
   2. Bilgrami AL
   3. Ahsan J

   (2022) Drosophila melanogaster chemosensory pathways as potential targets to curb the insect menace

   Insects 13:142.

   https://doi.org/10.3390/insects13020142

   • PubMed
   • Google Scholar
2. 1. Anderson DJ

   (2016) Circuit modules linking internal states and social behaviour in flies and mice

   Nature Reviews. Neuroscience 17:692–704.

   https://doi.org/10.1038/nrn.2016.125

   • PubMed
   • Google Scholar
3. 1. Antony C
   2. Davis TL
   3. Carlson DA
   4. Pechine JM
   5. Jallon JM

   (1985) Compared behavioral responses of male Drosophila melanogaster (Canton S) to natural and synthetic aphrodisiacs

   Journal of Chemical Ecology 11:1617–1629.

   https://doi.org/10.1007/BF01012116

   • PubMed
   • Google Scholar
4. 1. Bartelt RJ
   2. Schaner AM
   3. Jackson LL

   (1985) cis-Vaccenyl acetate as an aggregation pheromone in Drosophila melanogaster

   Journal of Chemical Ecology 11:1747–1756.

   https://doi.org/10.1007/BF01012124

   • PubMed
   • Google Scholar
5. Book

   1. Becker SD
   2. Hurst JL

   (2008) Pregnancy block from a female perspective in

   In: Hurst JL, Beynon RJ, Roberts SC, Wyatt TD, editors. Chemical Signals in Vertebrates 11. New York, NY: Springer. pp. 141–150.

   https://doi.org/10.1007/978-0-387-73945-8_13

   • Google Scholar
6. 1. Billeter JC
   2. Rideout EJ
   3. Dornan AJ
   4. Goodwin SF

   (2006) Control of male sexual behavior in Drosophila by the sex determination pathway

   Current Biology 16:R766–R776.

   https://doi.org/10.1016/j.cub.2006.08.025

   • PubMed
   • Google Scholar
7. 1. Billeter JC
   2. Atallah J
   3. Krupp JJ
   4. Millar JG
   5. Levine JD

   (2009) Specialized cells tag sexual and species identity in Drosophila melanogaster

   Nature 461:987–991.

   https://doi.org/10.1038/nature08495

   • PubMed
   • Google Scholar
8. 1. Billeter JC
   2. Levine JD

   (2015) The role of cVA and the Odorant binding protein Lush in social and sexual behavior in Drosophila melanogaster

   Frontiers in Ecology and Evolution 3:75.

   https://doi.org/10.3389/fevo.2015.00075

   • Google Scholar
9. 1. Bogovic JA
   2. Otsuna H
   3. Heinrich L
   4. Ito M
   5. Jeter J
   6. Meissner G
   7. Nern A
   8. Colonell J
   9. Malkesman O
   10. Ito K
   11. Saalfeld S

   (2020) An unbiased template of the Drosophila brain and ventral nerve cord

   PLOS ONE 15:e0236495.

   https://doi.org/10.1371/journal.pone.0236495

   • PubMed
   • Google Scholar
10. 1. Bontonou G
    2. Denis B
    3. Wicker-Thomas C

    (2013) Interaction between temperature and male pheromone in sexual isolation in Drosophila melanogaster

    Journal of Evolutionary Biology 26:2008–2020.

    https://doi.org/10.1111/jeb.12206

    • PubMed
    • Google Scholar
11. 1. Bruce HM

    (1959) An exteroceptive block to pregnancy in the mouse

    Nature 184:105.

    https://doi.org/10.1038/184105a0

    • PubMed
    • Google Scholar
12. 1. Bussell JJ
    2. Yapici N
    3. Zhang SX
    4. Dickson BJ
    5. Vosshall LB

    (2014) Abdominal-B neurons control Drosophila virgin female receptivity

    Current Biology 24:1584–1595.

    https://doi.org/10.1016/j.cub.2014.06.011

    • PubMed
    • Google Scholar
13. 1. Chiu H
    2. Robie AA
    3. Branson KM
    4. Vippa T
    5. Epstein S
    6. Rubin GM
    7. Anderson DJ
    8. Schretter CE

    (2023) Cell type-specific contributions to a persistent aggressive internal state in female Drosophila

    eLife 01:88598.

    https://doi.org/10.7554/eLife.88598.2

    • Google Scholar
14. Preprint

    1. Clements J
    2. Goina C
    3. Hubbard PM
    4. Kawase T
    5. Olbris DJ
    6. Otsuna H
    7. Svirskas R
    8. Rokicki K

    (2022) Neuronbridge: an intuitive web application for neuronal morphology search across large data sets

    bioRxiv.

    https://doi.org/10.1101/2022.07.20.500311

    • Google Scholar
15. 1. Couto A
    2. Alenius M
    3. Dickson BJ

    (2005) Molecular, anatomical, and functional organization of the Drosophila olfactory system

    Current Biology 15:1535–1547.

    https://doi.org/10.1016/j.cub.2005.07.034

    • PubMed
    • Google Scholar
16. 1. Davies NB

    (1983) Polyandry, cloaca-pecking and sperm competition in dunnocks

    Nature 302:334–336.

    https://doi.org/10.1038/302334a0

    • Google Scholar
17. 1. Dean R
    2. Nakagawa S
    3. Pizzari T

    (2011) The risk and intensity of sperm ejection in female birds

    The American Naturalist 178:343–354.

    https://doi.org/10.1086/661244

    • PubMed
    • Google Scholar
18. 1. Deutsch D
    2. Pacheco D
    3. Encarnacion-Rivera L
    4. Pereira T
    5. Fathy R
    6. Clemens J
    7. Girardin C
    8. Calhoun A
    9. Ireland E
    10. Burke A
    11. Dorkenwald S
    12. McKellar C
    13. Macrina T
    14. Lu R
    15. Lee K
    16. Kemnitz N
    17. Ih D
    18. Castro M
    19. Halageri A
    20. Jordan C
    21. Silversmith W
    22. Wu J
    23. Seung HS
    24. Murthy M

    (2020) The neural basis for a persistent internal state in Drosophila females

    eLife 9:e59502.

    https://doi.org/10.7554/eLife.59502

    • Google Scholar
19. Book

    1. Dixson AF

    (1998) Primate Sexuality: Comparative Studies of the Prosimians, Monkeys, Apes, and Human Beings

    Oxford University Press.

    https://doi.org/10.1093/acprof:osobl/9780199544646.001.0001

    • Google Scholar
20. 1. Duménil C
    2. Woud D
    3. Pinto F
    4. Alkema JT
    5. Jansen I
    6. Van Der Geest AM
    7. Roessingh S
    8. Billeter J-C

    (2016) Pheromonal cues deposited by mated females convey social information about egg-laying sites in Drosophila melanogaster

    Journal of Chemical Ecology 42:259–269.

    https://doi.org/10.1007/s10886-016-0681-3

    • PubMed
    • Google Scholar
21. 1. Dweck HKM
    2. Ebrahim SAM
    3. Thoma M
    4. Mohamed AAM
    5. Keesey IW
    6. Trona F
    7. Lavista-Llanos S
    8. Svatoš A
    9. Sachse S
    10. Knaden M
    11. Hansson BS

    (2015) Pheromones mediating copulation and attraction in Drosophila

    PNAS 112:E2829–E2835.

    https://doi.org/10.1073/pnas.1504527112

    • PubMed
    • Google Scholar
22. 1. Everaerts C
    2. Farine JP
    3. Cobb M
    4. Ferveur JF

    (2010) Drosophila cuticular hydrocarbons revisited: mating status alters cuticular profiles

    PLOS ONE 5:e9607.

    https://doi.org/10.1371/journal.pone.0009607

    • PubMed
    • Google Scholar
23. 1. Gaspar M
    2. Dias S
    3. Vasconcelos ML

    (2022) Mating pair drives aggressive behavior in female Drosophila

    Current Biology 32:4734–4742.

    https://doi.org/10.1016/j.cub.2022.09.009

    • PubMed
    • Google Scholar
24. 1. Grillet M
    2. Dartevelle L
    3. Ferveur JF

    (2006) A Drosophila male pheromone affects female sexual receptivity

    Proceedings. Biological Sciences 273:315–323.

    https://doi.org/10.1098/rspb.2005.3332

    • PubMed
    • Google Scholar
25. 1. Hamada FN
    2. Rosenzweig M
    3. Kang K
    4. Pulver SR
    5. Ghezzi A
    6. Jegla TJ
    7. Garrity PA

    (2008) An internal thermal sensor controlling temperature preference in Drosophila

    Nature 454:217–220.

    https://doi.org/10.1038/nature07001

    • Google Scholar
26. 1. Han C
    2. Peng Q
    3. Sun M
    4. Jiang X
    5. Su X
    6. Chen J
    7. Ma M
    8. Zhu H
    9. Ji X
    10. Pan Y

    (2022) The doublesex gene regulates dimorphic sexual and aggressive behaviors in Drosophila

    PNAS 119:2201513119.

    https://doi.org/10.1073/pnas.2201513119

    • Google Scholar
27. 1. Inagaki HK
    2. Panse KM
    3. Anderson DJ

    (2014) Independent, reciprocal neuromodulatory control of sweet and bitter taste sensitivity during starvation in Drosophila

    Neuron 84:806–820.

    https://doi.org/10.1016/j.neuron.2014.09.032

    • PubMed
    • Google Scholar
28. Software

    1. Jefferis G

    (2015) Fiji-cmtk-gui, version 0.1.1

    GitHub.

    https://github.com/jefferis/fiji-cmtk-gui
29. 1. Jiang X
    2. Pan Y

    (2022) Neural control of action selection among innate behaviors

    Neuroscience Bulletin 38:1541–1558.

    https://doi.org/10.1007/s12264-022-00886-x

    • PubMed
    • Google Scholar
30. 1. Joseph RM
    2. Carlson JR

    (2015) Drosophila chemoreceptors: a molecular interface between the chemical world and the brain

    Trends in Genetics 31:683–695.

    https://doi.org/10.1016/j.tig.2015.09.005

    • PubMed
    • Google Scholar
31. 1. Kim DW
    2. Yao Z
    3. Graybuck LT
    4. Kim TK
    5. Nguyen TN
    6. Smith KA
    7. Fong O
    8. Yi L
    9. Koulena N
    10. Pierson N
    11. Shah S
    12. Lo L
    13. Pool AH
    14. Oka Y
    15. Pachter L
    16. Cai L
    17. Tasic B
    18. Zeng H
    19. Anderson DJ

    (2019) Multimodal analysis of cell types in a hypothalamic node controlling social behavior

    Cell 179:713–728.

    https://doi.org/10.1016/j.cell.2019.09.020

    • Google Scholar
32. Preprint

    1. Kim YJ
    2. Lee KM
    3. Kim SJ
    4. Park SH
    5. Song I
    6. Yun M
    7. Zhang C
    8. Dus M
    9. Kim JI
    10. Lee S
    11. Suh GSB
    12. Park ZY
    13. Yang T

    (2023) Galactoside in the Male Ejaculate Evaluated as a Nuptial Gift by the Female Nutrient Sensing Neurons

    Research Square.

    https://doi.org/10.21203/rs.3.rs-2137467/v1

    • Google Scholar
33. 1. Kim DH
    2. Jang YH
    3. Yun M
    4. Lee KM
    5. Kim YJ

    (2024) Long-term neuropeptide modulation of female sexual drive via the TRP channel in Drosophila melanogaster

    PNAS 121:e2310841121.

    https://doi.org/10.1073/pnas.2310841121

    • PubMed
    • Google Scholar
34. 1. Klapoetke NC
    2. Murata Y
    3. Kim SS
    4. Pulver SR
    5. Birdsey-Benson A
    6. Cho YK
    7. Morimoto TK
    8. Chuong AS
    9. Carpenter EJ
    10. Tian Z
    11. Wang J
    12. Xie Y
    13. Yan Z
    14. Zhang Y
    15. Chow BY
    16. Surek B
    17. Melkonian M
    18. Jayaraman V
    19. Constantine-Paton M
    20. Wong GK-S
    21. Boyden ES

    (2014) Independent optical excitation of distinct neural populations

    Nature Methods 11:338–346.

    https://doi.org/10.1038/nmeth.2836

    • Google Scholar
35. 1. Koene JM
    2. Ter Maat A

    (2007) Coolidge effect in pond snails: male motivation in a simultaneous hermaphrodite

    BMC Evolutionary Biology 7:212.

    https://doi.org/10.1186/1471-2148-7-212

    • PubMed
    • Google Scholar
36. 1. Kohatsu S
    2. Koganezawa M
    3. Yamamoto D

    (2011) Female contact activates male-specific interneurons that trigger stereotypic courtship behavior in Drosophila

    Neuron 69:498–508.

    https://doi.org/10.1016/j.neuron.2010.12.017

    • PubMed
    • Google Scholar
37. 1. Kohatsu S
    2. Yamamoto D

    (2015) Visually induced initiation of Drosophila innate courtship-like following pursuit is mediated by central excitatory state

    Nature Communications 6:6457.

    https://doi.org/10.1038/ncomms7457

    • PubMed
    • Google Scholar
38. 1. Kohl J
    2. Huoviala P
    3. Jefferis GS

    (2015) Pheromone processing in Drosophila

    Current Opinion in Neurobiology 34:149–157.

    https://doi.org/10.1016/j.conb.2015.06.009

    • PubMed
    • Google Scholar
39. 1. Kohlmeier P
    2. Zhang Y
    3. Gorter JA
    4. Su CY
    5. Billeter JC

    (2021) Mating increases Drosophila melanogaster females’ choosiness by reducing olfactory sensitivity to a male pheromone

    Nature Ecology & Evolution 5:1165–1173.

    https://doi.org/10.1038/s41559-021-01482-4

    • PubMed
    • Google Scholar
40. 1. Kurtovic A
    2. Widmer A
    3. Dickson BJ

    (2007) A single class of olfactory neurons mediates behavioural responses to A Drosophila sex pheromone

    Nature 446:542–546.

    https://doi.org/10.1038/nature05672

    • PubMed
    • Google Scholar
41. 1. Laturney M
    2. Billeter JC

    (2016) Drosophila melanogaster females restore their attractiveness after mating by removing male anti-aphrodisiac pheromones

    Nature Communications 7:12322.

    https://doi.org/10.1038/ncomms12322

    • PubMed
    • Google Scholar
42. 1. Lee G
    2. Hall JC
    3. Park JH

    (2002) Doublesex gene expression in the central nervous system of Drosophila melanogaster

    Journal of Neurogenetics 16:229–248.

    https://doi.org/10.1080/01677060216292

    • PubMed
    • Google Scholar
43. 1. Lee KM
    2. Daubnerová I
    3. Isaac RE
    4. Zhang C
    5. Choi S
    6. Chung J
    7. Kim YJ

    (2015) A neuronal pathway that controls sperm ejection and storage in female Drosophila

    Current Biology 25:790–797.

    https://doi.org/10.1016/j.cub.2015.01.050

    • PubMed
    • Google Scholar
44. 1. Leinders-Zufall T
    2. Brennan P
    3. Widmayer P
    4. Shivalingappa PC
    5. Maul-Pavicic A
    6. Jäger M
    7. Li XH
    8. Breer H
    9. Zufall F
    10. Boehm T

    (2004) MHC class I peptides as chemosensory signals in the vomeronasal organ

    Science 306:1033–1037.

    https://doi.org/10.1126/science.1102818

    • PubMed
    • Google Scholar
45. 1. Lin CC
    2. Potter CJ

    (2015) Re-Classification of Drosophila melanogaster trichoid and intermediate sensilla using fluorescence-guided single sensillum recording

    PLOS ONE 10:e0139675.

    https://doi.org/10.1371/journal.pone.0139675

    • PubMed
    • Google Scholar
46. 1. Lin CC
    2. Prokop-Prigge KA
    3. Preti G
    4. Potter CJ

    (2015) Food odors trigger Drosophila males to deposit a pheromone that guides aggregation and female oviposition decisions

    eLife 4:e08688.

    https://doi.org/10.7554/eLife.08688

    • PubMed
    • Google Scholar
47. 1. Lin HH
    2. Cao DS
    3. Sethi S
    4. Zeng Z
    5. Chin JSR
    6. Chakraborty TS
    7. Shepherd AK
    8. Nguyen CA
    9. Yew JY
    10. Su CY
    11. Wang JW

    (2016) Hormonal modulation of pheromone detection enhances male courtship success

    Neuron 90:1272–1285.

    https://doi.org/10.1016/j.neuron.2016.05.004

    • PubMed
    • Google Scholar
48. 1. Liu T
    2. Wang Y
    3. Tian Y
    4. Zhang J
    5. Zhao J
    6. Guo A

    (2020) The receptor channel formed by ppk25, ppk29 and ppk23 can sense the Drosophila female pheromone 7,11-heptacosadiene

    Genes Brain Behav 19:e12529.

    https://doi.org/10.1111/gbb.12529

    • Google Scholar
49. 1. Liu M
    2. Kim DW
    3. Zeng H
    4. Anderson DJ

    (2022) Make war not love: The neural substrate underlying a state-dependent switch in female social behavior

    Neuron 110:841–856.

    https://doi.org/10.1016/j.neuron.2021.12.002

    • PubMed
    • Google Scholar
50. 1. Lone SR
    2. Sharma VK

    (2012) Or47b receptor neurons mediate sociosexual interactions in the fruit fly Drosophila melanogaster

    Journal of Biological Rhythms 27:107–116.

    https://doi.org/10.1177/0748730411434384

    • PubMed
    • Google Scholar
51. 1. Lone SR
    2. Venkataraman A
    3. Srivastava M
    4. Potdar S
    5. Sharma VK

    (2015) Or47b-neurons promote male-mating success in Drosophila

    Biology Letters 11:20150292.

    https://doi.org/10.1098/rsbl.2015.0292

    • PubMed
    • Google Scholar
52. 1. Lu B
    2. LaMora A
    3. Sun Y
    4. Welsh MJ
    5. Ben-Shahar Y

    (2012) ppk23-Dependent chemosensory functions contribute to courtship behavior in Drosophila melanogaster

    PLOS Genetics 8:e1002587.

    https://doi.org/10.1371/journal.pgen.1002587

    • PubMed
    • Google Scholar
53. 1. Mane SD
    2. Tompkins L
    3. Richmond RC

    (1983) Male esterase 6 catalyzes the synthesis of a sex pheromone in Drosophila melanogaster females

    Science 222:419–421.

    https://doi.org/10.1126/science.222.4622.419

    • PubMed
    • Google Scholar
54. 1. Manier MK
    2. Belote JM
    3. Berben KS
    4. Novikov D
    5. Stuart WT
    6. Pitnick S

    (2010) Resolving mechanisms of competitive fertilization success in Drosophila melanogaster

    Science 328:354–357.

    https://doi.org/10.1126/science.1187096

    • PubMed
    • Google Scholar
55. 1. Mohammad F
    2. Stewart JC
    3. Ott S
    4. Chlebikova K
    5. Chua JY
    6. Koh TW
    7. Ho J
    8. Claridge-Chang A

    (2017) Optogenetic inhibition of behavior with anion channelrhodopsins

    Nature Methods 14:271–274.

    https://doi.org/10.1038/nmeth.4148

    • PubMed
    • Google Scholar
56. Preprint

    1. Otsuna H
    2. Ito M
    3. Kawase T

    (2018) Color depth MIP mask search: a new tool to expedite split-gal4 creation

    bioRxiv.

    https://doi.org/10.1101/318006

    • Google Scholar
57. Software

    1. Otsuna H
    2. Kawase T
    3. Jefferis G

    (2020) ColorMIP_Mask_Search, version 1.0.1

    GitHub.

    https://github.com/JaneliaSciComp/ColorMIP_Mask_Search
58. 1. Parker GA

    (1970) Sperm competition and its evolutionary consequences in the insects

    Biological Reviews 45:525–567.

    https://doi.org/10.1111/j.1469-185X.1970.tb01176.x

    • Google Scholar
59. 1. Pizzari T
    2. Birkhead TR

    (2000) Female feral fowl eject sperm of subdominant males

    Nature 405:787–789.

    https://doi.org/10.1038/35015558

    • PubMed
    • Google Scholar
60. 1. Roberts EK
    2. Lu A
    3. Bergman TJ
    4. Beehner JC

    (2012) A Bruce effect in wild geladas

    Science 335:1222–1225.

    https://doi.org/10.1126/science.1213600

    • PubMed
    • Google Scholar
61. 1. Sato K
    2. Yamamoto D

    (2020) Contact-chemosensory evolution underlying reproductive isolation in Drosophila species

    Frontiers in Behavioral Neuroscience 14:597428.

    https://doi.org/10.3389/fnbeh.2020.597428

    • PubMed
    • Google Scholar
62. 1. Scheffer LK
    2. Xu CS
    3. Januszewski M
    4. Lu Z
    5. Takemura S-Y
    6. Hayworth KJ
    7. Huang GB
    8. Shinomiya K
    9. Maitlin-Shepard J
    10. Berg S
    11. Clements J
    12. Hubbard PM
    13. Katz WT
    14. Umayam L
    15. Zhao T
    16. Ackerman D
    17. Blakely T
    18. Bogovic J
    19. Dolafi T
    20. Kainmueller D
    21. Kawase T
    22. Khairy KA
    23. Leavitt L
    24. Li PH
    25. Lindsey L
    26. Neubarth N
    27. Olbris DJ
    28. Otsuna H
    29. Trautman ET
    30. Ito M
    31. Bates AS
    32. Goldammer J
    33. Wolff T
    34. Svirskas R
    35. Schlegel P
    36. Neace E
    37. Knecht CJ
    38. Alvarado CX
    39. Bailey DA
    40. Ballinger S
    41. Borycz JA
    42. Canino BS
    43. Cheatham N
    44. Cook M
    45. Dreher M
    46. Duclos O
    47. Eubanks B
    48. Fairbanks K
    49. Finley S
    50. Forknall N
    51. Francis A
    52. Hopkins GP
    53. Joyce EM
    54. Kim S
    55. Kirk NA
    56. Kovalyak J
    57. Lauchie SA
    58. Lohff A
    59. Maldonado C
    60. Manley EA
    61. McLin S
    62. Mooney C
    63. Ndama M
    64. Ogundeyi O
    65. Okeoma N
    66. Ordish C
    67. Padilla N
    68. Patrick CM
    69. Paterson T
    70. Phillips EE
    71. Phillips EM
    72. Rampally N
    73. Ribeiro C
    74. Robertson MK
    75. Rymer JT
    76. Ryan SM
    77. Sammons M
    78. Scott AK
    79. Scott AL
    80. Shinomiya A
    81. Smith C
    82. Smith K
    83. Smith NL
    84. Sobeski MA
    85. Suleiman A
    86. Swift J
    87. Takemura S
    88. Talebi I
    89. Tarnogorska D
    90. Tenshaw E
    91. Tokhi T
    92. Walsh JJ
    93. Yang T
    94. Horne JA
    95. Li F
    96. Parekh R
    97. Rivlin PK
    98. Jayaraman V
    99. Costa M
    100. Jefferis GS
    101. Ito K
    102. Saalfeld S
    103. George R
    104. Meinertzhagen IA
    105. Rubin GM
    106. Hess HF
    107. Jain V
    108. Plaza SM

    (2020) A connectome and analysis of the adult Drosophila central brain

    eLife 9:e57443.

    https://doi.org/10.7554/eLife.57443

    • PubMed
    • Google Scholar
63. 1. Schneider MR
    2. Mangels R
    3. Dean MD

    (2016) The molecular basis and reproductive function(s) of copulatory plugs

    Molecular Reproduction and Development 83:755–767.

    https://doi.org/10.1002/mrd.22689

    • PubMed
    • Google Scholar
64. 1. Schretter CE
    2. Aso Y
    3. Robie AA
    4. Dreher M
    5. Dolan MJ
    6. Chen N
    7. Ito M
    8. Yang T
    9. Parekh R
    10. Branson KM
    11. Rubin GM

    (2020) Cell types and neuronal circuitry underlying female aggression in Drosophila

    eLife 9:e58942.

    https://doi.org/10.7554/eLife.58942

    • PubMed
    • Google Scholar
65. 1. Schröder-Lang S
    2. Schwärzel M
    3. Seifert R
    4. Strünker T
    5. Kateriya S
    6. Looser J
    7. Watanabe M
    8. Kaupp UB
    9. Hegemann P
    10. Nagel G

    (2007) Fast manipulation of cellular cAMP level by light in vivo

    Nature Methods 4:39–42.

    https://doi.org/10.1038/nmeth975

    • PubMed
    • Google Scholar
66. 1. Scott D
    2. Richmond RC

    (1988) A genetic analysis of male-predominant pheromones in Drosophila melanogaster

    Genetics 119:639–646.

    https://doi.org/10.1093/genetics/119.3.639

    • PubMed
    • Google Scholar
67. Book

    1. Sengupta S
    2. Smith DP

    (2014)

    How Drosophila detect volatile pheromones: signaling, circuits behavior

    In: Mucignat-Caretta C, editors. Neurobiology of Chemical Communication, Frontiers in Neuroscience. Boca Raton (FL): CRC Press/Taylor & Francis. pp. 229–254.

    • Google Scholar
68. 1. Steiger S
    2. Franz R
    3. Eggert AK
    4. Müller JK

    (2008) The Coolidge effect, individual recognition and selection for distinctive cuticular signatures in a burying beetle

    Proceedings. Biological Sciences 275:1831–1838.

    https://doi.org/10.1098/rspb.2008.0375

    • PubMed
    • Google Scholar
69. 1. Sweeney ST
    2. Broadie K
    3. Keane J
    4. Niemann H
    5. O’Kane CJ

    (1995) Targeted expression of tetanus toxin light chain in Drosophila specifically eliminates synaptic transmission and causes behavioral defects

    Neuron 14:341–351.

    https://doi.org/10.1016/0896-6273(95)90290-2

    • PubMed
    • Google Scholar
70. 1. Tanenhaus AK
    2. Zhang J
    3. Yin JCP

    (2012) In vivo circadian oscillation of dCREB2 and NF-κB activity in the Drosophila nervous system

    PLOS ONE 7:e45130.

    https://doi.org/10.1371/journal.pone.0045130

    • PubMed
    • Google Scholar
71. 1. Thistle R
    2. Cameron P
    3. Ghorayshi A
    4. Dennison L
    5. Scott K

    (2012) Contact chemoreceptors mediate male-male repulsion and male-female attraction during Drosophila courtship

    Cell 149:1140–1151.

    https://doi.org/10.1016/j.cell.2012.03.045

    • PubMed
    • Google Scholar
72. 1. Toda H
    2. Zhao X
    3. Dickson BJ

    (2012) The Drosophila female aphrodisiac pheromone activates ppk23(+) sensory neurons to elicit male courtship behavior

    Cell Reports 1:599–607.

    https://doi.org/10.1016/j.celrep.2012.05.007

    • PubMed
    • Google Scholar
73. 1. van der Goes van Naters W
    2. Carlson JR

    (2007) Receptors and neurons for fly odors in Drosophila

    Current Biology 17:606–612.

    https://doi.org/10.1016/j.cub.2007.02.043

    • Google Scholar
74. 1. Verschut TA
    2. Ng R
    3. Doubovetzky NP
    4. Le Calvez G
    5. Sneep JL
    6. Minnaard AJ
    7. Su CY
    8. Carlsson MA
    9. Wertheim B
    10. Billeter JC

    (2023) Aggregation pheromones have a non-linear effect on oviposition behavior in Drosophila melanogaster

    Nature Communications 14:1544.

    https://doi.org/10.1038/s41467-023-37046-2

    • PubMed
    • Google Scholar
75. 1. Vijayan V
    2. Thistle R
    3. Liu T
    4. Starostina E
    5. Pikielny CW

    (2014) Drosophila pheromone-sensing neurons expressing the ppk25 ion channel subunit stimulate male courtship and female receptivity

    PLOS Genetics 10:e1004238.

    https://doi.org/10.1371/journal.pgen.1004238

    • PubMed
    • Google Scholar
76. 1. Wagner RH
    2. Helfenstein F
    3. Danchin E

    (2004) Female choice of young sperm in a genetically monogamous bird

    Proceedings. Biological Sciences 271 Suppl 4:S134–S137.

    https://doi.org/10.1098/rsbl.2003.0142

    • PubMed
    • Google Scholar
77. 1. Wang L
    2. Han X
    3. Mehren J
    4. Hiroi M
    5. Billeter JC
    6. Miyamoto T
    7. Amrein H
    8. Levine JD
    9. Anderson DJ

    (2011) Hierarchical chemosensory regulation of male-male social interactions in Drosophila

    Nature Neuroscience 14:757–762.

    https://doi.org/10.1038/nn.2800

    • PubMed
    • Google Scholar
78. 1. Wang F
    2. Wang K
    3. Forknall N
    4. Patrick C
    5. Yang T
    6. Parekh R
    7. Bock D
    8. Dickson BJ

    (2020) Neural circuitry linking mating and egg laying in Drosophila females

    Nature 579:101–105.

    https://doi.org/10.1038/s41586-020-2055-9

    • PubMed
    • Google Scholar
79. 1. Wang Z
    2. Receveur JP
    3. Pu J
    4. Cong H
    5. Richards C
    6. Liang M
    7. Chung H

    (2022) Desiccation resistance differences in Drosophila species can be largely explained by variations in cuticular hydrocarbons

    eLife 11:e80859.

    https://doi.org/10.7554/eLife.80859

    • Google Scholar
80. 1. Wei D
    2. Talwar V
    3. Lin D

    (2021) Neural circuits of social behaviors: Innate yet flexible

    Neuron 109:1600–1620.

    https://doi.org/10.1016/j.neuron.2021.02.012

    • PubMed
    • Google Scholar
81. 1. Werthebach M
    2. Stewart FA
    3. Gahlen A
    4. Mettler-Altmann T
    5. Akhtar I
    6. Maas-Enriquez K
    7. Droste A
    8. Eichmann TO
    9. Poschmann G
    10. Stühler K
    11. Beller M

    (2019) Control of Drosophila growth and survival by the lipid droplet-associated protein CG9186/Sturkopf

    Cell Reports 26:3726–3740.

    https://doi.org/10.1016/j.celrep.2019.02.110

    • PubMed
    • Google Scholar
82. 1. Yagound B
    2. Blacher P
    3. Chameron S
    4. Châline N

    (2012) Social context and reproductive potential affect worker reproductive decisions in a eusocial insect

    PLOS ONE 7:e52217.

    https://doi.org/10.1371/journal.pone.0052217

    • PubMed
    • Google Scholar
83. 1. Yapici N
    2. Kim YJ
    3. Ribeiro C
    4. Dickson BJ

    (2008) A receptor that mediates the post-mating switch in Drosophila reproductive behaviour

    Nature 451:33–37.

    https://doi.org/10.1038/nature06483

    • PubMed
    • Google Scholar
84. 1. Yu JY
    2. Kanai MI
    3. Demir E
    4. Jefferis G
    5. Dickson BJ

    (2010) Cellular organization of the neural circuit that drives Drosophila courtship behavior

    Current Biology 20:1602–1614.

    https://doi.org/10.1016/j.cub.2010.08.025

    • PubMed
    • Google Scholar
85. 1. Yu KE
    2. Kim DH
    3. Kim YI
    4. Jones WD
    5. Lee JE

    (2018) Mass spectrometry-based screening platform reveals orco interactome in Drosophila melanogaster

    Molecules and Cells 41:150–159.

    https://doi.org/10.14348/molcells.2018.2305

    • PubMed
    • Google Scholar
86. 1. Zhou C
    2. Pan Y
    3. Robinett CC
    4. Meissner GW
    5. Baker BS

    (2014) Central brain neurons expressing doublesex regulate female receptivity in Drosophila

    Neuron 83:149–163.

    https://doi.org/10.1016/j.neuron.2014.05.038

    • PubMed
    • Google Scholar
87. 1. Zhuang L
    2. Sun Y
    3. Hu M
    4. Wu C
    5. La X
    6. Chen X
    7. Feng Y
    8. Wang X
    9. Hu Y
    10. Xue L

    (2016) Or47b plays a role in Drosophila males’ preference for younger mates

    Open Biology 6:160086.

    https://doi.org/10.1098/rsob.160086

    • PubMed
    • Google Scholar
88. 1. Zipple MN
    2. Roberts EK
    3. Alberts SC
    4. Beehner JC

    (2019) Male-mediated prenatal loss: functions and mechanisms

    Evolutionary Anthropology 28:114–125.

    https://doi.org/10.1002/evan.21776

    • PubMed
    • Google Scholar

Author details

1. Minsik Yun

   School of Life Sciences, Gwangju Institute of Science and Technology (GIST), Gwangju, Republic of Korea

   Contribution

   Conceptualization, Investigation, Writing – original draft

   Competing interests

   No competing interests declared

   "This ORCID iD identifies the author of this article:" 0000-0002-0011-0942

2. Do-Hyoung Kim

   School of Life Sciences, Gwangju Institute of Science and Technology (GIST), Gwangju, Republic of Korea

   Contribution

   Investigation

   Competing interests

   No competing interests declared

3. Tal Soo Ha

   Department of Biomedical Science, College of Natural Science, Daegu University, Gyeongsan, Republic of Korea

   Contribution

   Investigation

   Competing interests

   No competing interests declared

   "This ORCID iD identifies the author of this article:" 0000-0003-0015-6075

4. Kang-Min Lee

   School of Life Sciences, Gwangju Institute of Science and Technology (GIST), Gwangju, Republic of Korea

   Contribution

   Methodology

   Competing interests

   No competing interests declared

5. Eungyu Park

   School of Life Sciences, Gwangju Institute of Science and Technology (GIST), Gwangju, Republic of Korea

   Contribution

   Investigation

   Competing interests

   No competing interests declared

6. Markus Knaden

   1. Department of Evolutionary Neuroethology, Max Planck Institute for Chemical Ecology, Jena, Germany
   2. Next Generation Insect Chemical Ecology, Max Planck Centre, Max Planck Institute for Chemical Ecology, Jena, Germany

   Contribution

   Methodology, Writing – review and editing

   Competing interests

   No competing interests declared

   "This ORCID iD identifies the author of this article:" 0000-0002-6710-1071

7. Bill S Hansson

   1. Department of Evolutionary Neuroethology, Max Planck Institute for Chemical Ecology, Jena, Germany
   2. Next Generation Insect Chemical Ecology, Max Planck Centre, Max Planck Institute for Chemical Ecology, Jena, Germany

   Contribution

   Methodology, Writing – review and editing

   Competing interests

   No competing interests declared

   "This ORCID iD identifies the author of this article:" 0000-0002-4811-1223

8. Young-Joon Kim

   School of Life Sciences, Gwangju Institute of Science and Technology (GIST), Gwangju, Republic of Korea

   Contribution

   Conceptualization, Supervision, Funding acquisition, Writing – original draft, Writing – review and editing

   For correspondence

   kimyj@gist.ac.kr

   Competing interests

   No competing interests declared

   "This ORCID iD identifies the author of this article:" 0000-0002-7990-754X

• 745

  views

• 75

  downloads

• 0

  citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.