Home Journals Articles Updates About
CN
image
收藏切换
Sex-Specific and State-Dependent Neuromodulation Regulates Male and Female Locomotion and Sexual Behaviors
收藏切换
PDF
Xinyu Jiang1, Mengshi Sun1, Jie Chen1, Yufeng Pan1, 2, *
Research. Vol 7 Article ID 0321
Less
收藏切换
Research. Vol 7 Article ID 0321
• Rapid Report •
Sex-Specific and State-Dependent Neuromodulation Regulates Male and Female Locomotion and Sexual Behaviors
Full
Xinyu Jiang1, Mengshi Sun1, Jie Chen1, Yufeng Pan1, 2, *
Affiliations
  • 1The Key Laboratory of Developmental Genes and Human Disease, School of Life Science and Technology, Southeast University, Nanjing 210096, China.
  • 2Co-innovation Center of Neuroregeneration, Nantong University, Nantong 226019, China.
Published: 2024-02-22 doi: 10.34133/research.0321
Outline
收藏切换

Males and females display dimorphic behaviors that often involve sex-specific locomotor patterns. How the sexually dimorphic locomotion is mediated is poorly understood. In this study, we identify a neuropeptide that oppositely regulates locomotion for efficient sexual behaviors in Drosophila males and females. We find that males are less active than females if isolated. However, when sexually aroused through activating homologous but sexually dimorphic pC1 neurons, males exhibit higher activity levels than females. We discover diuretic hormone 44 (DH44) that functions in pC1 neurons in a sex-specific way to inhibit male locomotion and promote female locomotion. Surprisingly, DH44 exerts opposite effects in sexually aroused flies to promote male locomotion and suppress female locomotion, which is crucial for successful male courtship and female receptivity. These findings demonstrate sexually dimorphic and state-dependent control of locomotor activity by pC1 neuronal activity and DH44 modulation.

Xinyu Jiang, Mengshi Sun, Jie Chen, Yufeng Pan. Sex-Specific and State-Dependent Neuromodulation Regulates Male and Female Locomotion and Sexual Behaviors[J]. Research, 2024 , 7 (2) : 0321 . DOI: 10.34133/research.0321
Full Text
Less
Introduction
Less
A long-standing question in neuroscience is how sexually dimorphic behaviors are generated. In both invertebrates and vertebrates, a few quantitative differences of the nervous system, especially in the interneurons, are found in the 2 sexes of the same species establishing the neural basis of sexually dimorphic behaviors [1]. In Drosophila, the neuronal substrates underlying sexual dimorphism are specified by the male-specific product encoded by fruitless and the sex-specific products encoded by doublesex (DsxM in males and DsxF in females) [2–4]. A subset of interneurons defined by dsx in the posterior brain region termed pC1 in both sexes have been found to control dimorphic sexual and aggressive behaviors [5–9]. There are ~60 DsxM-positive pC1 neurons in males, but only ~6 DsxF-positive counterparts in females [10,11]. These pC1 neurons integrate multiple sex-related sensory inputs and encode internal states in both males and females [7,10,12]. The huge differences in the numbers and arborizations of pC1 neurons in males and females provide the circuit basis underlying sexually dimorphic behaviors. Here, we discover a neuromodulatory mechanism for sexual dimorphism, in which a neuropeptide functions in the established dimorphic pC1 neurons to specify dimorphic locomotor activities in males and females for efficient sexual behaviors.
Results and Discussion
Less
To compare locomotor activity in male and female flies, we firstly assayed spontaneous locomotion in 2 wild-type strains (WTCS and w1118) for 24 hours. We found that isolated males displayed significantly lower locomotor activity than isolated females (Fig. 1A). We next set out to compare locomotor differences in sexually aroused males and females by artificially activating sex-promoting pC1 neurons. We used an intersectional strategy as done previously (dsxGAL4 and R71G01-LexA) [8,13] to label homologous pC1 neurons in both sexes of the same genotype. Such intersection labeled ~23 neurons per hemisphere in the male brain and ~6 neurons per hemisphere in the female brain, hereafter referred to as pC1R71G01 (Fig. 1B). We firstly expressed the Drosophila temperature-sensitive cation channel dTrpA1 in pC1R71G01 neurons for activation and set the temperatures as 5 gradients from 22 to 32 °C, where 22 °C represented the baseline. While control males consistently showed lower activity than control females at various temperatures, males with pC1R71G01 neurons being activated displayed significantly higher activity than females with activated pC1R71G01 neurons at each temperature (Fig. 1C and D and Fig. S1A and B).
Because of the lower precisions in the time resolution of thermogenetic protocols, we further drove the expression of red-shifted channel CsChrimson in pC1R71G01 neurons for optogenetic activation using a constant red light with 9 increasing intensities (level 1 to 9; Fig. 1E). We found that moderate activation (level 4) of pC1R71G01 neurons increased male locomotion and decreased female locomotion such that males are more active than females (Fig. 1F), which is generally consistent with the dTrpA1-mediated activation experiments. However, we also found distinct effects during and after optogenetic activation. While strong activation (>level 4) promoted both male and female locomotion to comparable levels during the “ON” phases (Fig. 1F and H), we observed a remarkably enhancement of male locomotion but a reduction of female locomotion following the light “OFF” (Fig. 1G and H), which may reflect distinct internal states in males and females. We propose that the long-term dTrpA1 activation may correspond to a summed effect of both “ON” and “OFF” phases from optogenetic activation.
To investigate whether the regular activity of pC1 neurons is involved in locomotor control, we silenced pC1 neurons by expressing the inwardly rectifying potassium channel Kir2.1 and observed similarly low levels of locomotion in both males and females (Fig. 1I and Fig. S1C and D). Together, these experiments demonstrate that pC1 neurons play crucial roles in regulating the sexually dimorphic locomotor activity in both sexually isolated and aroused males and females (Fig. 1J).
To identify molecular markers for pC1 neurons, we isolated male pC1 neurons for RNA sequencing and found that a neuropeptide-encoding gene diuretic hormone 44 (DH44) was highly expressed in these cells (Fig. 2A). Intersection of the 2 knockin drivers, dsxGAL4 and DH44LexA, labeled ~6 pairs of pC1 neurons in males and ~2 pairs in females (Fig. 2B and D). pC1 neurons were also labeled from intersection of 2 other knockin drivers, dsxLexA and DH44GAL4 (Fig. S2A and B), as well as a split GAL4 of DH44AD and dsxDBD (Fig. S2C and D). Double staining with anti-DH44 antibody confirmed that at least 4 of the 6 labeled male cells and the 2 female cells indeed colocalized with DH44 (Fig. 2C and E).
We next used RNA interference against DH44 to investigate its potential role in regulating male and female locomotion. The effectiveness of the UAS-DH44IR line was verified by the lethal effect during the pupal stage under the control of actin-GAL4 and the negative immunostaining signal with anti-DH44 using the DH44GAL4 driver (Fig. S3). Interestingly, we observed a sex-specific effect of DH44. Knocking down DH44 driven by dsxGAL4 increased male locomotion but decreased female locomotion such that these males and females showed indistinguishable levels of locomotion (Fig. 2F and G). In contrast, knocking down choline acetyltransferase (ChAT) that encodes the biosynthetic enzyme for acetylcholine in dsxGAL4 neurons reduced male and female locomotion to similarly low levels (Fig. S4A). Given that strong activation of pC1R71G01 neurons dramatically increased locomotion in both sexes, we wondered how DH44 was involved. To our surprise, after knocking down DH44 in dsx neurons at a state of activating pC1R71G01 neurons via dTrpA1, male flies locomoted much more slowly, while female flies locomoted much faster (Fig. 2H and I). These data reveal that the already high locomotor activity resulting from intense activation of pC1R71G01 neurons is promoted by DH44 in males but suppressed by DH44 in females. These results indicate a state-dependent function of DH44, in addition to its sex-specific function.
To further confirm the above findings, we optogenetically activated pC1R71G01 neurons (intensity level 7) for 1 min in solitary males or females and continued to record locomotion for 10 min after light went off. As expected, pC1R71G01>CsChrimson males showed a robust increase in locomotor activity during photostimulation and went much higher after light offset. Furthermore, males with additional dsxGAL4-driven knockdown of DH44 showed higher activity in baseline but no enhancement during photostimulation stage and a much less enhancement during the 2-min stage after photostimulation (Fig. 2J and K). In contrast, pC1R71G01>CsChrimson females moved slightly faster during photostimulation and returned to the baseline levels within a short time after light offset. However, DH44-knockdown females showed a lower locomotor activity in baseline, a higher activity during activation and decayed rapidly after it (Fig. 2J and K). Consequently, knockdown of DH44 driven by dsxGAL4 significantly reduced both male courtship (Fig. 2L) and female receptivity (Fig. 2M). Moreover, knocking down DH44 specifically in pC1 neurons, but not vpoDN neurons crucial for vaginal plate opening, dramatically reduced female receptivity (Fig. S5).
Together, these results indicate that DH44 inhibits male locomotion but promotes female locomotion when they are sexually isolated, but, inversely, DH44 promotes male locomotion but inhibits female locomotion when they are sexually aroused with pC1 activation (Fig. 2N). The persistently heightened locomotor activity observed in males, but not females, following pC1 activation may indicate distinct internal states encoded by the sexually dimorphic pC1 neurons, which may be beneficial for both male and female reproductive behaviors. We speculate that DH44 functions in a subset of pC1 neurons, and perhaps in other neurons too, to play sex-specific and state-dependent modulatory roles via its 2 receptors. In most animal species, males command active locomotion to follow and court females with an elaborate ritual, while females are more passive in such a ritual and often reduce locomotion when deciding to accept a courting male. The male-specific and state-dependent modulation of locomotion by DH44 makes males more active and sensitive to potential mates, while saving energy without them. In contrast, the female-specific and state-dependent regulation by DH44 promotes locomotion if not being courted, probably for food and/or oviposition searching, and reduces locomotion for copulation if satisfied with male courtship.
It is worth noting that functional counterparts for both the DH44 peptide (corticotropin-releasing factor) and pC1 neurons (ventrolateral subdivision of the ventromedial hypothalamus) exist in mammals [14,15]. Thus, the sex-specific and state-dependent neuromodulation we discovered in flies could also apply to high-order animals. Future studies would reveal how DH44 exerts the sex-specific and state-dependent functions in pC1 neurons in molecular and cellular details to coordinate male and female innate behaviors.
Materials and Methods
Less
Flies were raised in a 22 or 25 °C environment with 60% humidity and a 12-h:12-h light:dark cycle. All the tested flies were group-housed virgins (∼20 single-sex flies per vial) between the ages of 5 to 7 d after eclosion without specifically mentioned. Canton-S and w1118 flies were used as wild-type strains. Detailed methods, full genotypes of tested flies, data analysis were described in the Supplementary Materials.
Funding
Less
  • Key Technologies Research and Development Program (2021YFA1101300)
  • National Natural Science Foundation of China (32371067; 31970943)
References
Less
1.
Yang CF, Shah NM. Representing sex in the brain, one module at a time. Neuron. 2014;82(2):261–278.
2.
Burtis KC, Baker BS. Drosophila doublesex gene controls somatic sexual differentiation by producing alternatively spliced mRNAs encoding related sex-specific polypeptides. Cell. 1989;56(6):997–1010.
3.
Ito H, Fujitani K, Usui K, Shimizu-Nishikawa K, Tanaka S, Yamamoto D. Sexual orientation in drosophila is altered by the satori mutation in the sex-determination gene fruitless that encodes a zinc finger protein with a BTB domain. Proc Natl Acad Sci U S A. 1996;93(18):9687–9692.
4.
Ryner LC, Goodwin SF, Castrillon DH, Anand A, Villella A, Baker BS, Hall JC, Taylor BJ, Wasserman SA. Control of male sexual behavior and sexual orientation in Drosophila by the fruitless gene. Cell. 1996;87(6):1079–1089.
5.
Palavicino-Maggio CB, Chan YB, McKellar C, Kravitz EA. A small number of cholinergic neurons mediate hyperaggression in female Drosophila. Proc Natl Acad Sci U S A. 2019;116(34):17029–17038.
6.
Han C, Peng Q, Sun M, Jiang X, Su X, Chen J, Ma M, Zhu H, Ji X, Pan Y. The doublesex gene regulates dimorphic sexual and aggressive behaviors in Drosophila. Proc Natl Acad Sci U S A. 2022;119(37): Article e2201513119.
7.
Hoopfer ED, Jung Y, Inagaki HK, Rubin GM, Anderson DJ. P1 interneurons promote a persistent internal state that enhances inter-male aggression in Drosophila. elife. 2015;4:e11346.
8.
Zhou C, Pan Y, Robinett CC, Meissner GW, Baker BS. Central brain neurons expressing doublesex regulate female receptivity in Drosophila. Neuron. 2014;83(1):149–163.
9.
Wan X, Shen P, Shi K, Li J, Wu F, Zhou C. A neural circuit controlling virgin female aggression induced by mating-related cues in Drosophila. Neurosci Bull. 2023;39(9):1396–1410.
10.
Jiang X, Pan Y. Neural control of action selection among innate behaviors. Neurosci Bull. 2022;38(12):1541–1558.
11.
Asahina K. Sex differences in Drosophila behavior: Qualitative and quantitative dimorphism. Curr Opin Physiol. 2018;6:35–45.
12.
Deutsch D, Pacheco D, Encarnacion-Rivera L, Pereira T, Fathy R, Clemens J, Girardin C, Calhoun A, Ireland E, Burke A, et al. The neural basis for a persistent internal state in Drosophila females. elife. 2020;9:e59502.
13.
Pan Y, Meissner GW, Baker BS. Joint control of Drosophila male courtship behavior by motion cues and activation of male-specific P1 neurons. Proc Natl Acad Sci U S A. 2012;109(25):10065–10070.
14.
Anderson DJ. Circuit modules linking internal states and social behaviour in flies and mice. Nat Rev Neurosci. 2016;17(11):692–704.
15.
Cabrero P, Radford JC, Broderick KE, Costes L, Veenstra JA, Spana EP, Davies SA, Dow JAT. The Dh gene of Drosophila melanogaster encodes a diuretic peptide that acts through cyclic AMP. J Exp Biol. 2002;205(Pt 24):3799–3807.
Appendix
Less
Year 2024 volume 7 Issue 2
PDF
183
103
Cite this Article
BibTeX
Article Info
doi: 10.34133/research.0321
  • Receive Date：2023-12-03
  • Online Date：2025-07-24
  • Published：2024-02-22
Article Data
Affiliations
History
  • Received：2023-12-03
  • Accepted：2024-01-26
Funding
Key Technologies Research and Development Program (2021YFA1101300)
National Natural Science Foundation of China (32371067; 31970943)
Affiliations
    1The Key Laboratory of Developmental Genes and Human Disease, School of Life Science and Technology, Southeast University, Nanjing 210096, China.
    2Co-innovation Center of Neuroregeneration, Nantong University, Nantong 226019, China.

Corresponding:

* Address correspondence to:
References
Share
https://castjournals.cast.org.cn/joweb/research/EN/10.34133/research.0321
Share to
QR

Scan QR to access full text

Cite this article
BibTeX
Citations
表12种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
关闭全屏
  • BibTeX
  • EndNote
  • RefWorks
  • TxT
Links
Articles
Latest Articles
Most Read
Collections
Updates
Events
News
Multimedia
About
About Us
Contact
No. 86 Xueyuan South Road, Haidian District, Beijing
100081
010-62199257
qkjq@cast.org.cn

Copyright © 2025 China Association for Science and Technology. All rights reserved. For all open access content, the relevant licensing terms apply.
Sponsored by the Office of the Leading Group for Cybersecurity and Informatization of CAST, and supported by Science and Technology Review Publishing House