P65 Targets FGFR1 to Regulate the Survival of Ovarian Granulosa Cells

Yuan, Xiaolong; Li, Zhonghui; Kong, Yaru; Zhong, Yuyi; He, Yingting; Zhang, Ailing; Zhou, Xiaofeng; Jiang, Yao; Zhang, Zhe; Zhang, Hao; Li, Jiaqi

doi:10.3390/cells8111334

Cited by 11 publications

(9 citation statements)

References 52 publications

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“…It is well known that transcription factors modulate gene expression by interacting with the promoter regions of related genes. For example, p65 may target the −348/−338 region of fibroblast growth factor receptor 1 ( FGFR1 ) to promote the transcription of FGFR1 and enhance the pro-proliferation and anti-apoptotic effect of FGFR1 to facilitate the growth of follicles [ 33 ]. Thus, to explore effects of rs81471943 on the expression of EXOC4 , the SNP markers which link with rs81471943 at the promoter of EXOC4 were further investigated by constructing 5′-deletions for EXOC4 .…”

Section: Discussionmentioning

confidence: 99%

The Association of an SNP in the EXOC4 Gene and Reproductive Traits Suggests Its Use as a Breeding Marker in Pigs

Zhou

Zheng

et al. 2021

Animals

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In mammals, the exocyst complex component 4 (EXOC4) gene has often been reported to be involved in vesicle transport. The SNP rs81471943 (C/T) is located in the intron of porcine EXOC4, while six quantitative trait loci (QTL) within 5–10 Mb around EXOC4 are associated with ovary weight, teat number, total offspring born alive, and corpus luteum number. However, the molecular mechanisms between EXOC4 and the reproductive performance of pigs remains to be elucidated. In this study, rs81471943 was genotyped from a total of 994 Duroc sows, and the genotype and allele frequency of SNP rs81471943 (C/T) were statistically analyzed. Then, the associations between SNP rs81471943 and four reproductive traits, including number of piglets born alive (NBA), litter weight at birth (LWB), number of piglets weaned (NW), and litter weight at weaning (LWW), were determined. Sanger sequencing and PCR restriction fragment length polymorphism (PCR-RFLP) were utilized to identify the rs81471943 genotype. We found that the genotype frequency of CC was significantly higher than that of CT and TT, and CC was the most frequent genotype for NBA, LWB, NW, and LWW. Moreover, 5′-deletion and luciferase assays identified a positive transcription regulatory element in the EXOC4 promoter. After exploring the EXOC4 promoter, SNP -1781G/A linked with SNP rs81471943 (C/T) were identified by analysis of the transcription activity of the haplotypes, and SNP -1781 G/A may influence the potential binding of P53, E26 transformation specific sequence -like 1 transcription factor (ELK1), and myeloid zinc finger 1 (MZF1). These findings provide useful information for identifying a molecular marker of EXOC4-assisted selection in pig breeding.

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Section: Discussionmentioning

confidence: 99%

The Association of an SNP in the EXOC4 Gene and Reproductive Traits Suggests Its Use as a Breeding Marker in Pigs

Zhou

Zheng

et al. 2021

Animals

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“…The RNA extraction and qPCR were performed as previously described (Yuan et al, 2018; Yuan et al, 2019). The expression level of glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) mRNA was used as internal reference, and the relative mRNA expression levels of the target genes were calculated by relative quantification (2 −ΔΔct ).…”

Section: Methodsmentioning

confidence: 99%

“…The culture of porcine ovarian GCs was performed according to our previously published methods (Yuan et al, 2018; Yuan et al, 2019). Briefly, the porcine ovaries from prepubertal gilts were collected from a local slaughterhouse for the commercial pigs (Duroc × Landrace × Yorkshire) in Guangzhou and transferred to our laboratory in phosphate‐buffered saline (PBS) containing penicillin (100 IU/ml) and streptomycin (100 μg/ml) (Invitrogen, Shanghai, China).…”

Section: Methodsmentioning

confidence: 99%

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FoxA2 and p53 regulate the transcription of HSD17B1 in ovarian granulosa cells of pigs

Yuan

Zhou

Qiao

et al. 2020

Reprod Domestic Animals

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The oestrogens have been highly implicated in the fertility of female animals. It is widely known that the oestrogens are primarily synthetized by the ovarian granulosa cells (GCs), and the final and essential step of this process is to catalyse the oestrone to the more active oestradiol by the protein coded by hydroxysteroid 17‐beta dehydrogenase 1 (HSD17B1) gene. However, the molecular mechanism regarding the transcription of HSD17B1 remains to be fully elucidated in ovarian GCs. In this study, the 5’‐deletion, luciferase assay and chromatin immunoprecipitation (ChIP) were utilized to explore the molecular regulation of transcription of HSD17B1 with the porcine ovarian GCs as the cellular model. After the deletions with −2105 to −1754 bp, −1753 to −1429 bp, −1430 to −1081 bp and −1082 to −730 bp, the relative luciferase activity of HSD17B1 promoter did not change significantly, but the deletion of −731 to −332 bp significantly increased the relative luciferase activity of HSD17B1 promoter, and an insertion (GTTT) that might raise the transcription of HSD17B1 was identified at −401 bp of HSD17B1. These findings suggested the region from −731 to +38 bp was the core promoter of HSD17B1, and the region between −731 to −332 bp might be a silence element for HSD17B1. Furthermore, the forkhead box A2 (FoxA2) directly bound at −412 to −401 bp to negatively but p53 bound at −383 to −374 bp to positively regulate the transcription and translation of HSD17B1 in ovarian GCs. These findings will improve our understanding on HSD17B1‐mediated oestrogens and provide useful information for further investigations into fertility of females.

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“…The culture medium was refreshed after 48 h. The oocytes were washed away because they were suspended in the medium. The pGCs were used in further research when they reached 90%-100% confluency at 24-48 h. The culture of pGCs was performed following the methods of reference (Yuan et al, 2019).…”

Section: Porcine Granulosa Cells Culturementioning

confidence: 99%

MIR143 Inhibits Steroidogenesis and Induces Apoptosis Repressed by H3K27me3 in Granulosa Cells

Zhong

Chen

et al. 2020

Front. Cell Dev. Biol.

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The granulosa cell growth factor and apoptotic factor are two factors to determine follicular apoptosis. Whether ssc-miR-143-3p (MIR143) plays as an apoptosis factor in porcine granulosa cells (pGCs) remain unclear. This study tries to investigate what function of MIR143 is and how MIR143 gets these functions in pGCs from 3 to 5 mm medium-sized follicles. Firstly, 5 RACE was used to identify the structure of MIR143, and in situ hybridization, qPCR, and DNA pull-down were employed to exhibit the spatio-temporal expression and transcriptional regulation of MIR143. Furthermore, ELISA, Western blotting, and flow cytometry were adopted to explore the functions of MIR143 in pGCs. It was found that MIR143 was an exonic miRNA located in host gene LOC100514340 with an increasing expression during follicular growth. Moreover, MIR143 suppressed steroidogenesis related genes of HSD17β4, ER1, and PTGS2, negatively regulating estrogen, androgen, progesterone, and prostaglandin. MIR143 induced the apoptosis via activation of BAX-dependent Caspase 3 signaling. Furthermore, H3K27me3 influenced the recruitment of transcription factors and binding proteins to repress MIR143 transcription. At last, H3K27me3 agonist with MIR143 inhibition activated steroidogenesis but repressed apoptosis. These findings suggest that H3K27me3-mediated MIR143 inhibition play a critical role in follicular atresia by regulating cell apoptosis and steroidogenesis, which will provide useful information for further investigations of H3K27me3-miediated MIR143 epigenetic regulation in follicular growth in mammals.

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P65 Targets FGFR1 to Regulate the Survival of Ovarian Granulosa Cells

Cited by 11 publications

References 52 publications

The Association of an SNP in the EXOC4 Gene and Reproductive Traits Suggests Its Use as a Breeding Marker in Pigs

The Association of an SNP in the EXOC4 Gene and Reproductive Traits Suggests Its Use as a Breeding Marker in Pigs

FoxA2 and p53 regulate the transcription of HSD17B1 in ovarian granulosa cells of pigs

MIR143 Inhibits Steroidogenesis and Induces Apoptosis Repressed by H3K27me3 in Granulosa Cells

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