Chen Dong, Zhang Qi-Tong, Li Wei-Jun, Han Hong-Liang, Smagghe Guy, Yan Ying, Jiang Hong-Bo, Wang Jin-Jun, Wei Dong
Chongqing Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection, Southwest University, Chongqing, China.
International Joint Laboratory of China-Belgium on Sustainable Crop Pest Control, Academy of Agricultural Sciences, Southwest University, Chongqing, China.
Pest Manag Sci. 2025 Sep;81(9):5201-5213. doi: 10.1002/ps.8874. Epub 2025 May 15.
BACKGROUND: Insect spermatogenesis is a complex process. Numerous genes are involved in sperm motility, which is crucial for male fertility. Few long non-coding RNAs (lncRNAs) in the testis regulate insect spermatogenesis. We previously identified 364 testis-enriched lncRNAs in the globally invasive pest Zeugodacus cucurbitae Coquillett. One of these lncRNAs, lnc94641, is abundantly expressed in the testis; however, its role in spermatogenesis remains unknown. RESULTS: Suppression of lnc94641 expression led to a 60% decrease in spermatozoa count and a 29% decrease in offspring hatchability. A microRNA (miRNA), miR-957-3p, was experimentally demonstrated to bind to lnc94641 competitively. miR-957-3p overexpression recapitulated reproductive defect phenotypes similar to those caused by lnc94641 knockdown. Furthermore, target gene predictions combined with quantitative reverse transcription PCR, RNA pull-down, and dual luciferase reporter assays confirmed that miR-957-3p targets voltage-gated potassium channel 5 (VGKC5) and odorant receptor 85c (OR85c), elucidating a functional lncRNA-miRNA-mRNA competing endogenous RNA (ceRNA) regulatory axis. Fluorescence in situ hybridization (FISH) assays demonstrated the co-localization of lnc94641, miR-957-3p, and VGKC5/OR85c in the mature and transformed regions of the testes. Suppression of VGKC5/OR85c expression resulted in a 68% and 50% decrease in spermatozoa number and an 18% and 21% decrease in offspring hatchability, mirroring the phenotype observed with lnc94641-silencing, thereby reinforcing the mechanistic coherence of this regulatory network. CONCLUSION: These results revealed a ceRNA axis mediated by 'lnc94641-miR957-3p-VGKC5/OR85c' involved in spermatogenesis that impairs male fertility in the melon fly. Molecular perturbations (lncRNA knockdown or miRNA overexpression) consistently impair sperm production and offspring viability by dysregulating ion channels and chemosensory genes. This mechanistically resolved pathway, centered on the core components VGKC5 and OR85c, revealed conserved reproductive vulnerabilities that could enable the targeted genetic control of this agricultural pest. © 2025 Society of Chemical Industry.
背景：昆虫精子发生是一个复杂的过程。许多基因参与精子活力，而精子活力对雄性生育力至关重要。睾丸中很少有长链非编码RNA（lncRNA）调控昆虫精子发生。我们之前在全球入侵害虫瓜实蝇（Zeugodacus cucurbitae Coquillett）中鉴定出364个睾丸富集lncRNA。其中一个lncRNA，lnc94641，在睾丸中大量表达；然而，其在精子发生中的作用仍不清楚。 结果：lnc94641表达的抑制导致精子数量减少60%，后代孵化率降低29%。实验证明一种微小RNA（miRNA），miR-957-3p，能竞争性结合lnc94641。miR-957-3p的过表达重现了与lnc94641敲低引起的类似生殖缺陷表型。此外，结合定量逆转录PCR、RNA下拉和双荧光素酶报告基因检测的靶基因预测证实miR-957-3p靶向电压门控钾通道5（VGKC5）和气味受体85c（OR85c），阐明了一个功能性lncRNA-miRNA-mRNA竞争性内源RNA（ceRNA）调控轴。荧光原位杂交（FISH）检测证明lnc94641、miR-957-3p和VGKC5/OR85c在睾丸的成熟和转化区域共定位。VGKC5/OR85c表达的抑制导致精子数量分别减少68%和50%，后代孵化率分别降低18%和21%，这与lnc94641沉默时观察到的表型相似，从而加强了这个调控网络的机制连贯性。 结论：这些结果揭示了一个由“lnc94641-miR957-3p-VGKC5/OR85c”介导的参与精子发生的ceRNA轴，该轴损害了瓜实蝇的雄性生育力。分子扰动（lncRNA敲低或miRNA过表达）通过失调离子通道和化学感应基因持续损害精子产生和后代活力。这个以核心成分VGKC5和OR85c为中心的机制解析途径揭示了保守的生殖脆弱性，这可能有助于对这种农业害虫进行靶向基因控制。© 2025化学工业协会。
Int J Mol Sci. 2025-6-17
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