# GAL4-based functional screen of neuropeptides in Drosophila reproduction

**Authors:** Madhumala K. Sadanandappa, Caliope Marin, Shinae Park, Shivaprasad H. Sathyanarayana, Giovanni Bosco, Md Rajib Sharker, Md Rajib Sharker, Md Rajib Sharker

PMC · DOI: 10.1371/journal.pone.0345918 · PLOS One · 2026-03-27

## TL;DR

This study identifies 14 neuropeptides in fruit flies that affect reproduction, including eight that were previously unknown to play a role in this process.

## Contribution

A novel functional screening framework for identifying neuropeptides involved in Drosophila reproduction.

## Key findings

- Disruption of 14 neuropeptides altered egg-laying in Drosophila.
- Eight neuropeptides were newly identified as involved in reproductive regulation.
- The study provides a curated resource for future mechanistic studies on brain-gonad communication.

## Abstract

Neuropeptides are evolutionarily conserved signaling molecules that regulate diverse behavioral and physiological processes, including reproduction. Although, several neuropeptides have established roles in reproductive regulation, the reproductive functions of many neuropeptides in Drosophila melanogaster remain poorly characterized. Here, we performed a targeted neurogenetic screening to systematically assess the contribution of 25 neuropeptides to reproductive output. Using neuropeptide-specific GAL4 drivers and synaptic silencing with tetanus toxin, we quantified the egg-laying as an integrated functional readout of reproduction. Disruption of 14 neuropeptides altered egg-laying, including eight neuropeptides not previously described to play roles in reproductive regulation. While some of these effects are likely indirect and may reflect contributions from both female and male flies or systematic physiological signaling, these results reveal broad involvement of neuropeptidergic pathways in reproductive function. Collectively, this study establishes a functional screening framework, identifies new reproductive neuropeptides, and provides a curated resource to guide future mechanistic studies of neuropeptide-mediated brain-gonad communication.

## Linked entities

- **Species:** Drosophila melanogaster (taxon 7227)

## Full-text entities

- **Genes:** Dh44 (Diuretic hormone 44) [NCBI Gene 41170] {aka CG8348, CRF, DH, DH 44, DH-44, DH[[44]]}, AstA (Allatostatin A) [NCBI Gene 42947] {aka ALLS, AS, ASA, AST, AST-1, AST-3}, Dh31 (Diuretic hormone 31) [NCBI Gene 34181] {aka CG13094, DH 31, DH-II, DH[31], DH[[31], DH[[31]]}, ETH (Ecdysis triggering hormone) [NCBI Gene 44125] {aka CG18105, DmETH, Dmel\CG18105, Drm-ETH-1, Drm-ETH-2, ETH-1}, Eh (Eclosion hormone) [NCBI Gene 42101] {aka CG5400, DMEHAB, Dmel\CG5400, Drm-EH, EHAB}, fru (fruitless) [NCBI Gene 42226] {aka BTB-VI, BtbVI, CG14307, CG7688, CG7689, CG7690}, tnt (tent) [NCBI Gene 246802], CCK (cholecystokinin) [NCBI Gene 885], Akh (Adipokinetic hormone) [NCBI Gene 38495] {aka ADKH, AKH1, CG1171, Dmel\CG1171, Drm-AKH, DrmAKH}, SIFaR (SIFamide receptor) [NCBI Gene 42530] {aka CG10823, Dmel\CG10823, DrmSIFR, SIF-R, SIFR, SIFa-R}, PK1-R (Pyrokinin 1 receptor) [NCBI Gene 41713] {aka CG9918, Dmel\CG9918, LstR, PK1R, PK1r, Pk1R}, LGALS4 (galectin 4) [NCBI Gene 3960] {aka GAL4, L36LBP}, CCAP (Crustacean cardioactive peptide) [NCBI Gene 42683] {aka CG4910, DmCCAP, Dmel\CG4910}, Crz (Corazonin) [NCBI Gene 41742] {aka CG3302, COR, CORZ, Cora, Dm-Crz, Dmel\CG3302}, sNPF (short neuropeptide F precursor) [NCBI Gene 35286] {aka 38B.2, CG13968, Dmel\CG13968, Drm-sNPF, Drm-sNPF-1, Drm-sNPF-2}, Hug (Hugin) [NCBI Gene 41547] {aka CG6371, Dmel\CG6371, Drm-MT2, Drm-PK-2, HUGgamma, Hug-PK}, RYa (RYamide) [NCBI Gene 5740597] {aka CG40733, Dmel\CG40733, dRYamide}, CapaR (Capability receptor) [NCBI Gene 40393] {aka CAP2B-R, CAPA-R, CG14575, Capa R, Dm CG14575, Dmel\CG14575}, SP (Sex Peptide) [NCBI Gene 39494] {aka ACP, ACP70A, Acp-70A, Acp70, Acp70A, Acp70a}, Gal (beta galactosidase) [NCBI Gene 33839] {aka CG9092, DmelGal, Dmel\CG9092, beta-GAL, beta-Gal-1, beta-gal}, Dsk (Drosulfakinin) [NCBI Gene 45845] {aka BcDNA:RH46857, CCK, CG18090, DSK I, DSK II, DSK-0}, NPF (neuropeptide F) [NCBI Gene 42018] {aka CG10342, Dm-NPF, DmNPF, Dmel\CG10342, Drm-NPF, NP-PP}, Capa (Capability) [NCBI Gene 43541] {aka CAP, CAP-2b, CAP2B-1, CAP2B-2, CAP2B-3, CAP2B/pyrokinin}, anon-15Aa (anon-15Aa) [NCBI Gene 5657260] {aka gene x}, Burs (Bursicon) [NCBI Gene 42560] {aka Bur alpha, Burs alpha, Burs-alpha, BursA, Bursalpha, Bursicon alpha}, thick (thick) [NCBI Gene 45367] {aka tk}, InR (Insulin-like receptor) [NCBI Gene 42549] {aka 18402, CG18402, DIHR, DILR, DIR, DIRH}, Pdf (Pigment-dispersing factor) [NCBI Gene 43193] {aka BcDNA:RH08487, CG6496, Dmel\CG6496, Drm-PDF, Drm-PDH, Drm-pdf}, INS (insulin) [NCBI Gene 3630] {aka IDDM, IDDM1, IDDM2, ILPR, IRDN, MODY10}, SPR (Sex peptide receptor) [NCBI Gene 31463] {aka CG12731, CG16752, Dmel\CG16752, DrmSPR, SP-R}, AkhR (Adipokinetic hormone receptor) [NCBI Gene 33942] {aka AKH R, AKH-R, Akh-R, AkhR/GRHR, Akhr/GRHR, BEST:GH19447}, Ms (Myosuppressin) [NCBI Gene 44324] {aka CG6440, DMS, Dm-MS, Dmel\CG6440, Dms, Drm-MS}, Lgals4 (lectin, galactose binding, soluble 4) [NCBI Gene 16855] {aka gal-4}, PITRM1 (pitrilysin metallopeptidase 1) [NCBI Gene 10531] {aka MP1, PreP, SCAR30}, Syb (Synaptobrevin) [NCBI Gene 36080] {aka 21.15, CG12210, Dmel\CG12210, END1-2, J, VAMP}, CRH (corticotropin releasing hormone) [NCBI Gene 1392] {aka CRF, CRH1}, Lk (Leucokinin) [NCBI Gene 47746] {aka CG13480, DLK, Dmel\CG13480, Drm-KIN, Kinin, LCK}, Mip (Myoinhibiting peptide precursor) [NCBI Gene 39933] {aka ASB, AST-B, Ast-B, AstB, AstB-1, AstB/MIP}, Dh31-R (Diuretic hormone 31 Receptor) [NCBI Gene 36475] {aka CG17043, CG17415, CG32843, DH31, DH31-R1, DH31R}, CREB1 (cAMP responsive element binding protein 1) [NCBI Gene 1385] {aka CREB, CREB-1}, Pburs (Partner of Bursicon) [NCBI Gene 34845] {aka A, Bur beta, Burs beta, Bursbeta, CG15284, Dmel\CG15284}, AstC (Allatostatin C) [NCBI Gene 34537] {aka ASC, AST-C, Ast-C, Ast2, BEST:GH06087, BcDNA:RH36507}, SIFa (SIFamide) [NCBI Gene 3346202] {aka CG33527, Dmel\CG33527, Drm-IFa, Drm-SIFa, IFAMIDE, IFa}, CCHa1 (CCHamide-1) [NCBI Gene 41711] {aka CCH1, CCM, CG14358, Dm-CCHa1, Dmel\CG14358}, FMRFa (FMRFamide) [NCBI Gene 36030] {aka CG2346, DPKQDFMRF-amide, DPKQDFMRFamide, Dmel\CG2346, Drm-FMRF-1, Drm-FMRF-2}
- **Diseases:** egg-laying (MESH:D021181), aggression (MESH:D010554)
- **Chemicals:** agar (MESH:D000362), lipid (MESH:D008055), 4',6-diamidino-2-phenylindole (MESH:C007293), follicle-stimulating hormone (MESH:D005640), polyamine (MESH:D011073), phalloidin (MESH:D010590), Alexa fluor (-), luteinizing hormone (MESH:D007986), ecdysteroid (MESH:D026461), carbon dioxide (MESH:D002245), carbohydrate (MESH:D002241), ecdysone (MESH:D004440)
- **Species:** Drosophila melanogaster (fruit fly, species) [taxon 7227], Diptera (flies, order) [taxon 7147], Homo sapiens (human, species) [taxon 9606], Gallus gallus (bantam, species) [taxon 9031], Mus musculus (house mouse, species) [taxon 10090], Saccharomyces cerevisiae (baker's yeast, species) [taxon 4932], Sarcophaga bullata (grey fleshfly, species) [taxon 7385]

## Full text

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## Figures

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## References

232 references — full list in the complete paper: https://tomesphere.com/paper/PMC13028521/full.md

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Source: https://tomesphere.com/paper/PMC13028521