JOURNAL OF PROTEOME RESEARCH
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2022
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IF 5.37
Renzheng Zhang,Xian Guo,Chunnian Liang,Jie Pei,Pengjia Bao,Mancai Yin,Fude Wu,Min Chu,Ping Yan
To achieve fertilization, mammalian spermatozoa must undergo capacitation and the acrosome reaction (AR) within the female reproductive tract. However, the effects of cryopreservation on sperm maturation and fertilizing potential have yet to be established. To gain insight into changes in protein levels within sperm cells prepared for use in the context of fertilization, a comprehensive quantitative proteomic profiling approach was used to analyze frozen–thawed Ashidan yak spermatozoa under three sequential conditions: density gradient centrifugation-based purification, incubation in a capacitation medium, and treatment with the calcium ionophore A23187 to facilitate AR induction. In total, 3280 proteins were detected in these yak sperm samples, of which 3074 were quantified, with 68 and 32 being significantly altered following sperm capacitation and AR induction. Differentially abundant capacitation-related proteins were enriched in the metabolism and PPAR signaling pathways, while differentially abundant AR-related proteins were enriched in the AMPK signaling pathway. These data confirmed a role for superoxide dismutase 1 (SOD1) as a regulator of sperm capacitation while also offering indirect evidence that heat shock protein 90 alpha (HSP90AA1) regulates the AR. Together, these findings offer a means whereby sperm fertility-related marker proteins can be effectively identified. Data are available via Proteome Xchange with identifier PXD035038.
研究领域:
生殖生物学
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蛋白质组学
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动物科学
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产品关联
DOI
PubMed
高影响因子
Animals
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2024
·Vol. 14
·期 2
·240
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IF 2.7
Xudong Jiang,Yao Ma,Sanni Gong,Xiangdong Zi,Dawei Zhang
Simple SummaryThe growth and development of follicles is a prerequisite for normal fertility in female animals. Granulosa cells, as an important component of the ovary, play a key role in follicular development, and their apoptosis is an important cause of follicular atresia. Resveratrol, as a common antioxidant, has various biological properties in animals in vitro and in vivo. However, the functional role of resveratrol in many aspects of yak granulosa cell activity remains unclear. Thus, we evaluated the effects of resveratrol on yak granulosa cell activity in vitro. The results showed that resveratrol when added at an appropriate concentration to the culture media of yak granulosa cells in vitro, could promote proliferation, inhibit apoptosis, enhance antioxidant properties, and promote lipid metabolism and the progesterone secretion of yak granulosa cells. This study provides some theoretical basis for research to further improve yak reproduction.AbstractResveratrol (RES) is a class of natural polyphenolic compounds known for its strong anti-apoptotic and antioxidant properties. Granulosa cells (GCs) are one of the important components of ovarian follicles and play crucial roles in follicular development of follicles in the ovary. Here, we explored the effects of RES on the proliferation and functions of yak GCs. Firstly, we evaluated the effect of RES dose and time in culture on the viability of GCs, and then the optimum treatment protocol (10 μM RES, 36 h) was selected to analyze the effects of RES on the proliferation, cell cycle, apoptosis, malondialdehyde (MDA), glutathione (GSH), reactive oxygen species (ROS) accumulation, lipid droplet content, ATP production, and steroidogenesis of GCs, as well as the expression of related genes. The results show that RES treatment significantly (1) increased cell viability and proliferation and inhibited cell apoptosis by upregulatingBCL-2andSIRT1genes and downregulatingBAX,CASP3,P53, andKU70genes; (2) increased the proportion of GCs in the S phase and upregulatedCCND1,PCNA,CDK4, andCDK5genes; (3) reduced ROS accumulation and MDA content and increased GSH content, as well as upregulating the relative expression levels ofCAT,SOD2, andGPX1genes; (4) decreased lipid droplet content and increased ATP production; (5) promoted progesterone (P4) secretion and the expression of P4 synthesis-related genes (StAR,HSD3B1,andCYP11A1); and (6) inhibited E2 secretion andCYP19A1expression. These findings suggest that RES at 10 μM increases the proliferation and antioxidant properties, inhibits apoptosis, and promotes ATP production, lipid droplet consumption, and P4 secretion of yak GCs.Keywords:yak;resveratrol;granulosa cell;cell proliferation;antioxidant property;steroidogenesis
研究领域:
生殖生物学
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动物科学
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细胞生物学
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内分泌学
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DOI
PubMed
高影响因子
THERIOGENOLOGY
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2023
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IF 2.8
Xu-dong Jiang,Yu Liu,Jian-fei Wu,San-ni Gong,Yao Ma,Xiang-dong Zi
Supplementation with acetyl- l -carnitine (ALC) during in vitro maturation significantly improves the rates of oocyte cleavage and morula and blastocyst formation in sheep and buffalo ; however, the mode of action of ALC in improving oocyte competence is not completely understood. Therefore, the aim of this study was to investigate the effects of ALC on proliferation, antioxidant properties, lipid droplet accumulation and steroid hormone secretion in yak ( Bos grunniens ) granulosa cells (GCs). Yak GCs were identified using FSHR immunofluorescence. The cells were treated with different concentrations of ALC, cell proliferation was detected by cell counting kit-8, and the optimal concentration and treatment time were determined for subsequent experiments. Then, reactive oxygen species (ROS) were detected by a DCFH-DA probe, and lipid droplet accumulation was observed by oil red O staining. Estradiol (E2) and progesterone (P4) in the medium were detected by ELISA, and the expression of genes related to cell proliferation, apoptosis, the cell cycle, antioxidants and steroid synthesis was determined by RT‒qPCR. The results showed that 1 mM ALC treatment for 48 h was the optimum treatment. It significantly increased cell viability ( P < 0.05), significantly decreased the amount of ROS and lipid droplet content, and promoted P4 and E2 secretion ( P < 0.05) of yak GCs. RT‒qPCR results verified that GCs treated with 1 mM ALC for 48 h significantly increased the expression of genes related to anti-apoptosis and the cell cycle ( BCL-2 , PCNA , CCND1 and CCNB1 ), antioxidants ( CAT , SOD2 and GPX1 ), and E2 and P4 secretion ( StAR, CYP19A1 and HSD3B1 ) ( P < 0.05), but it significantly decreased the expression of apoptosis genes ( BAX and P53 ) ( P < 0.05). In conclusion, ALC increased the viability of yak GCs, reduced the amount of ROS and lipid droplets, increased P4 and E2 synthesis and affected the expression of related genes in yak GCs.
研究领域:
生殖生物学
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细胞生物学
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内分泌学
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分子生物学
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DOI
PubMed
高影响因子
ANIMAL BIOTECHNOLOGY
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2024
·
IF 1.7
Yueyue Li,Hongmei Xu,Zifeng Ma,Qiao Li,Yan Xiong,Xianrong Xiong,Jian Li,Daoliang Lan,Wei Fu
Apoptosis-inducing factor mitochondrion-associated 2 (AIFM2) has been identified as a gene with anti-ferroptosis properties. To explore whether AIFM2 exerts anti-ferroptosis role in yaks (Bos grunniens), we cloned yak AIFM2 gene and analyzed its biological characteristics. The coding region of AIFM2 had 1122 bp and encoded 373 amino acids, which was conserved in mammals. Next, RT-qPCR results showed an extensive expression of AIMF2 in yak tissues. Furthermore, we isolated yak skin fibroblasts (YSFs) and established a bisphenol A (BPA)-induced ferroptosis model to further investigate the role of AIFM2. BPA elevated oxidative stress (reactive oxygen species, ROS) and lipid peroxidation (malondialdehyde, MDA and BODIPY), and reduced cell viability and antioxidant capacity (glutathione, GSH), with the severity depending on the dosage. Of note, a supplement of Ferrostatin-1 (Fer), an inhibitor of ferroptosis, restored the previously mentioned indicators. Subsequently, we constructed an AIFM2 overexpression vector and designed AIFM2 specific interfering siRNAs, which were transfected into YSFs. The results showed that overexpressing AIFM2 alleviated ferroptosis, characterizing by significant changes of cell viability, ROS, BODIPY, MDA and GSH. Meanwhile, interfering AIFM2 aggravated ferroptosis, demonstrating the critical anti-ferroptosis role of the yak AIFM2 gene. This study shed light on further exploring the molecular mechanism of AIFM2 in plateau adaptability.
研究领域:
分子生物学
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遗传学
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细胞生物学
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毒理学
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DOI
高影响因子
MOLECULAR AND CELLULAR ENDOCRINOLOGY
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2026
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IF 3.4
Yanbing Lou,Tingting Yang,Chenglong Xia,Shijun Yang,Huidan Deng,Yanqiu Zhu,Jing Fang,Zhicai Zuo,Hongrui Guo
CCS and ATOX1 is a specific target of copper in hormone secretion of ovarian granulosa cells. • MEK1-ERK1/2 pathway is involved in copper-affected hormone secretion. • Copper affects hormones secretion through CCS and ATOX1 dependent MEK1-ERK1/2 pathway. Steroid hormone synthesis in granulosa cells (GCs) is crucial for normal follicular development and ovulation. Emerging evidence suggests that copper (Cu), a trace element, influences hormone secretion in yak granulosa cells (YGCs); however, the underlying mechanisms remain unclear. In this study, we demonstrated that Cu supplementation upregulates estradiol (E2) and progesterone (P4) secretion in YGCs through the activation of the MEK1-ERK1/2 signaling pathway. Our findings indicate that the Cu chaperones ATOX1 and CCS, but not the Cu-dependent enzyme LOX, are essential for Cu-induced MEK1-ERK1/2 activation and subsequent hormone secretion. Gene silencing of ATOX1 or CCS resulted in reduced MEK1-ERK1/2 phosphorylation and steroidogenesis, highlighting their regulatory roles in this pathway. Although LOX influenced hormone secretion, it did not affect MEK-ERK phosphorylation, suggesting a MEK-independent mechanism. These findings reveal that Cu regulates steroid hormone synthesis in YGCs via a CCS- and ATOX1-dependent MEK1–ERK1/2 pathway, providing new insights into the molecular basis of Cu-associated reproductive disorders in yaks. Download: Download high-res image (289KB) Download: Download full-size image
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DOI
PubMed
高影响因子
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