{"title":"Melatonin regulates rabbit sperm motility and kinematics via the MT1/PKC signaling pathway.","authors":"Chongchong Wang, Yanyan Zhang, Shiwen He, Biao Jiang, Jinglei Huang, Hui Peng","doi":"10.5713/ab.24.0593","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>Melatonin, a highly conserved molecule, is essential for various physiological functions. Research suggests that incorporating melatonin into semen extender is beneficial to livestock sperm. However, the effect of melatonin on rabbit sperm and the molecular mechanism of melatonin-regulated rabbit sperm motility and kinematics remains unclear. The objective of this study was to reveal the molecular mechanism that melatonin regulates rabbit sperm motility and kinematics.</p><p><strong>Methods: </strong>In this study, we investigated the expression and localization of melatonin-related proteins in rabbit testis and epididymis. Rabbit sperm was incubated under different concentration of melatonin for 60 min or 90 min at 37 °C, and then evaluated sperm motility parameters by IVOS II computer assisted sperm analyzer (CASA) system. Then we detected the sperm plasma membrane integrity, mitochondrial membrane potentials and intracellular reactive oxygen species levels. Furthermore, melatonin receptor antagonists were added to the extender and investigated the involvement of the melatonin receptors in the regulation of sperm motility parameters. Meanwhile, we carried out phosphoproteomics analysis and confirmed the regulation of rabbit sperm kinematics by melatonin via inhibition of the potential signaling pathway.</p><p><strong>Results: </strong>We found that melatonin-related proteins were expressed and localized in rabbit testes and the epididymis. Melatonin (5 mM) markedly increased rabbit sperm motility and kinematics as well as sperm quality. Furthermore, melatonin regulated rabbit sperm motility and kinematics via the MT1 receptor. Meanwhile, phosphoproteomics of rabbit sperm combined with GPS 5.0 software revealed the potential signaling pathway by which melatonin regulates sperm kinematics. Moreover, inhibition of PKC markedly reduced rabbit sperm kinematics, while inhibition of ERK1/2, p38 MAPK, PKG and JNKs with kinase inhibitors did not obviously change sperm kinematics. In addition, inhibition of the MT1 receptor significantly weakened PKC activity, suggesting that PKC is downstream of MT1.</p><p><strong>Conclusion: </strong>We therefore conclude that melatonin regulates rabbit sperm kinematics through the MT1/PKC signaling pathway.</p>","PeriodicalId":7825,"journal":{"name":"Animal Bioscience","volume":" ","pages":""},"PeriodicalIF":2.4000,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Animal Bioscience","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.5713/ab.24.0593","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"AGRICULTURE, DAIRY & ANIMAL SCIENCE","Score":null,"Total":0}
引用次数: 0
Abstract
Objective: Melatonin, a highly conserved molecule, is essential for various physiological functions. Research suggests that incorporating melatonin into semen extender is beneficial to livestock sperm. However, the effect of melatonin on rabbit sperm and the molecular mechanism of melatonin-regulated rabbit sperm motility and kinematics remains unclear. The objective of this study was to reveal the molecular mechanism that melatonin regulates rabbit sperm motility and kinematics.
Methods: In this study, we investigated the expression and localization of melatonin-related proteins in rabbit testis and epididymis. Rabbit sperm was incubated under different concentration of melatonin for 60 min or 90 min at 37 °C, and then evaluated sperm motility parameters by IVOS II computer assisted sperm analyzer (CASA) system. Then we detected the sperm plasma membrane integrity, mitochondrial membrane potentials and intracellular reactive oxygen species levels. Furthermore, melatonin receptor antagonists were added to the extender and investigated the involvement of the melatonin receptors in the regulation of sperm motility parameters. Meanwhile, we carried out phosphoproteomics analysis and confirmed the regulation of rabbit sperm kinematics by melatonin via inhibition of the potential signaling pathway.
Results: We found that melatonin-related proteins were expressed and localized in rabbit testes and the epididymis. Melatonin (5 mM) markedly increased rabbit sperm motility and kinematics as well as sperm quality. Furthermore, melatonin regulated rabbit sperm motility and kinematics via the MT1 receptor. Meanwhile, phosphoproteomics of rabbit sperm combined with GPS 5.0 software revealed the potential signaling pathway by which melatonin regulates sperm kinematics. Moreover, inhibition of PKC markedly reduced rabbit sperm kinematics, while inhibition of ERK1/2, p38 MAPK, PKG and JNKs with kinase inhibitors did not obviously change sperm kinematics. In addition, inhibition of the MT1 receptor significantly weakened PKC activity, suggesting that PKC is downstream of MT1.
Conclusion: We therefore conclude that melatonin regulates rabbit sperm kinematics through the MT1/PKC signaling pathway.
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