RFRP-3 Influences Apoptosis and Steroidogenesis of Yak Cumulus Cells and Compromises Oocyte Meiotic Maturation and Subsequent Developmental Competence
RF amide-related peptide 3 (RFRP-3), a mammalian ortholog of gonadotropin-inhibitory hormone (GnIH), is identified to be a novel inhibitory endogenous neurohormonal peptide that regulates mammalian reproduction by binding with specific G protein-coupled receptors (GPRs) in various species. Herein, o...
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MDPI AG
2023-04-01
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| author | Xianrong Xiong Yulei Hu Bangting Pan Yanjin Zhu Xixi Fei Qinhui Yang Yumian Xie Yan Xiong Daoliang Lan Wei Fu Jian Li |
| author_facet | Xianrong Xiong Yulei Hu Bangting Pan Yanjin Zhu Xixi Fei Qinhui Yang Yumian Xie Yan Xiong Daoliang Lan Wei Fu Jian Li |
| author_sort | Xianrong Xiong |
| collection | DOAJ |
| description | RF amide-related peptide 3 (RFRP-3), a mammalian ortholog of gonadotropin-inhibitory hormone (GnIH), is identified to be a novel inhibitory endogenous neurohormonal peptide that regulates mammalian reproduction by binding with specific G protein-coupled receptors (GPRs) in various species. Herein, our objectives were to explore the biological functions of exogenous RFRP-3 on the apoptosis and steroidogenesis of yak cumulus cells (CCs) and the developmental potential of yak oocytes. The spatiotemporal expression pattern and localization of GnIH/RFRP-3 and its receptor GPR147 were determined in follicles and CCs. The effects of RFRP-3 on the proliferation and apoptosis of yak CCs were initially estimated by EdU assay and TUNEL staining. We confirmed that high-dose (10<sup>−6</sup> mol/L) RFRP-3 suppressed viability and increased the apoptotic rates, implying that RFRP-3 could repress proliferation and induce apoptosis. Subsequently, the concentrations of E<sub>2</sub> and P<sub>4</sub> were significantly lower with 10<sup>−6</sup> mol/L RFRP-3 treatment than that of the control counterparts, which indicated that the steroidogenesis of CCs was impaired after RFRP-3 treatment. Compared with the control group, 10<sup>−6</sup> mol/L RFRP-3 treatment decreased the maturation of yak oocytes efficiently and subsequent developmental potential. We sought to explore the potential mechanism of RFRP-3-induced apoptosis and steroidogenesis, so we observed the levels of apoptotic regulatory factors and hormone synthesis-related factors in yak CCs after RFRP-3 treatment. Our results indicated that RFRP-3 dose-dependently elevated the expression of apoptosis markers (<i>Caspase</i> and <i>Bax</i>), whereas the expression levels of steroidogenesis-related factors (<i>LHR</i>, <i>StAR</i>, 3<i>β</i>-HSD) were downregulated in a dose-dependent manner. However, all these effects were moderated by cotreatment with inhibitory RF9 of GPR147. These results demonstrated that RFRP-3 adjusted the expression of apoptotic and steroidogenic regulatory factors to induce apoptosis of CCs, probably through binding with its receptor GPR147, as well as compromised oocyte maturation and developmental potential. This research revealed the expression profiles of GnIH/RFRP-3 and GPR147 in yak CCs and supported a conserved inhibitory action on oocyte developmental competence. |
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| spelling | doaj.art-859f7c693bbf4a149c2447a8a01e9a482023-11-17T19:33:40ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-04-01248700010.3390/ijms24087000RFRP-3 Influences Apoptosis and Steroidogenesis of Yak Cumulus Cells and Compromises Oocyte Meiotic Maturation and Subsequent Developmental CompetenceXianrong Xiong0Yulei Hu1Bangting Pan2Yanjin Zhu3Xixi Fei4Qinhui Yang5Yumian Xie6Yan Xiong7Daoliang Lan8Wei Fu9Jian Li10Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Exploitation of Ministry of Education, College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, ChinaKey Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Exploitation of Ministry of Education, College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, ChinaKey Laboratory for Animal Science of National Ethnic Affairs Commission, College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, ChinaKey Laboratory for Animal Science of National Ethnic Affairs Commission, College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, ChinaKey Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Exploitation of Ministry of Education, College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, ChinaKey Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Exploitation of Ministry of Education, College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, ChinaKey Laboratory for Animal Science of National Ethnic Affairs Commission, College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, ChinaKey Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Exploitation of Ministry of Education, College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, ChinaKey Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Exploitation of Ministry of Education, College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, ChinaKey Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Exploitation of Ministry of Education, College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, ChinaKey Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Exploitation of Ministry of Education, College of Animal & Veterinary Sciences, Southwest Minzu University, Chengdu 610041, ChinaRF amide-related peptide 3 (RFRP-3), a mammalian ortholog of gonadotropin-inhibitory hormone (GnIH), is identified to be a novel inhibitory endogenous neurohormonal peptide that regulates mammalian reproduction by binding with specific G protein-coupled receptors (GPRs) in various species. Herein, our objectives were to explore the biological functions of exogenous RFRP-3 on the apoptosis and steroidogenesis of yak cumulus cells (CCs) and the developmental potential of yak oocytes. The spatiotemporal expression pattern and localization of GnIH/RFRP-3 and its receptor GPR147 were determined in follicles and CCs. The effects of RFRP-3 on the proliferation and apoptosis of yak CCs were initially estimated by EdU assay and TUNEL staining. We confirmed that high-dose (10<sup>−6</sup> mol/L) RFRP-3 suppressed viability and increased the apoptotic rates, implying that RFRP-3 could repress proliferation and induce apoptosis. Subsequently, the concentrations of E<sub>2</sub> and P<sub>4</sub> were significantly lower with 10<sup>−6</sup> mol/L RFRP-3 treatment than that of the control counterparts, which indicated that the steroidogenesis of CCs was impaired after RFRP-3 treatment. Compared with the control group, 10<sup>−6</sup> mol/L RFRP-3 treatment decreased the maturation of yak oocytes efficiently and subsequent developmental potential. We sought to explore the potential mechanism of RFRP-3-induced apoptosis and steroidogenesis, so we observed the levels of apoptotic regulatory factors and hormone synthesis-related factors in yak CCs after RFRP-3 treatment. Our results indicated that RFRP-3 dose-dependently elevated the expression of apoptosis markers (<i>Caspase</i> and <i>Bax</i>), whereas the expression levels of steroidogenesis-related factors (<i>LHR</i>, <i>StAR</i>, 3<i>β</i>-HSD) were downregulated in a dose-dependent manner. However, all these effects were moderated by cotreatment with inhibitory RF9 of GPR147. These results demonstrated that RFRP-3 adjusted the expression of apoptotic and steroidogenic regulatory factors to induce apoptosis of CCs, probably through binding with its receptor GPR147, as well as compromised oocyte maturation and developmental potential. This research revealed the expression profiles of GnIH/RFRP-3 and GPR147 in yak CCs and supported a conserved inhibitory action on oocyte developmental competence.https://www.mdpi.com/1422-0067/24/8/7000RFRP-3apoptosissteroidogenesiscumulus celldevelopment |
| spellingShingle | Xianrong Xiong Yulei Hu Bangting Pan Yanjin Zhu Xixi Fei Qinhui Yang Yumian Xie Yan Xiong Daoliang Lan Wei Fu Jian Li RFRP-3 Influences Apoptosis and Steroidogenesis of Yak Cumulus Cells and Compromises Oocyte Meiotic Maturation and Subsequent Developmental Competence International Journal of Molecular Sciences RFRP-3 apoptosis steroidogenesis cumulus cell development |
| title | RFRP-3 Influences Apoptosis and Steroidogenesis of Yak Cumulus Cells and Compromises Oocyte Meiotic Maturation and Subsequent Developmental Competence |
| title_full | RFRP-3 Influences Apoptosis and Steroidogenesis of Yak Cumulus Cells and Compromises Oocyte Meiotic Maturation and Subsequent Developmental Competence |
| title_fullStr | RFRP-3 Influences Apoptosis and Steroidogenesis of Yak Cumulus Cells and Compromises Oocyte Meiotic Maturation and Subsequent Developmental Competence |
| title_full_unstemmed | RFRP-3 Influences Apoptosis and Steroidogenesis of Yak Cumulus Cells and Compromises Oocyte Meiotic Maturation and Subsequent Developmental Competence |
| title_short | RFRP-3 Influences Apoptosis and Steroidogenesis of Yak Cumulus Cells and Compromises Oocyte Meiotic Maturation and Subsequent Developmental Competence |
| title_sort | rfrp 3 influences apoptosis and steroidogenesis of yak cumulus cells and compromises oocyte meiotic maturation and subsequent developmental competence |
| topic | RFRP-3 apoptosis steroidogenesis cumulus cell development |
| url | https://www.mdpi.com/1422-0067/24/8/7000 |
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