INHBA transfection regulates proliferation, apoptosis and hormone synthesis in sheep granulosa cells

Bao, Yongjin; Yao, Xiaolei; Li, Xiaodan; EI-Samahy, M.A.; Yang, Hua; Liang, Yaxu; Liu, Zifei; Wang, Zhen

doi:10.1016/j.theriogenology.2021.09.004

Cited by 22 publications

(6 citation statements)

References 41 publications

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“…The IHC assay was performed as previously described [ 61 ]. Briefly, ovarian paraffin sections (six mm thickness) were rehydrated through graded ethanol series.…”

Section: Methodsmentioning

confidence: 99%

Neuromedin S Regulates Steroidogenesis through Maintaining Mitochondrial Morphology and Function via NMUR2 in Goat Ovarian Granulosa Cells

Sun

Cheng

Wang

et al. 2022

IJMS

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Neuromedin S (NMS) plays various roles in reproductive regulation, while the mechanism by which NMS regulates ovarian steroidogenesis remains unclear. In the current study, we confirmed the enhancement role of NMS in steroidogenesis in goat ovarian granulosa cells (GCs). To further explore the specific mechanism, we conducted a knockdown of NMUR2 in GCs followed by treatment with NMS and determined the effects of NMS treatment on mitochondrial morphology and function. The results found that NMS treatment increased the production of estrogen and up-regulated the expression of STAR, CYP11A1, 3BHSD, and CYP19A1, while the effects of NMS treatment were blocked by the knockdown of NMUR2 in goat GCs. Moreover, NMS treatment enhanced the fusion of mitochondria and up-regulated the expression of OPA1, MFN1, and MFN2, and increased mitochondrial membrane potential, the activity of respiratory chain enzymes and ATP production by maintaining a low expression level of mitochondrial unfolded protein response markers. The effects of NMS treatment on mitochondria were reversed by NMUR2 knockdown and NMS cotreatment. The possible mechanism of the results above was revealed by NMS treatment activating the Hippo pathway effector YAP1 and then managing the expression of phosphorylation PPARGC1A (Ser571). Together, these data showed that NMS promoted the fusion of mitochondria and protected mitochondrial function from mitochondrial unfolded protein response possibly via the NMUR2/YAP1/PPARGC1A pathway, thereby affecting the steroidogenesis of goat GCs. By elaborating the potential mechanism of NMS in regulating estrogen production in goat GCs, our results can serve as the mechanism reference for follicular growth and development.

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“…The IHC assay was performed as previously described [ 61 ]. Briefly, ovarian paraffin sections (six mm thickness) were rehydrated through graded ethanol series.…”

Section: Methodsmentioning

confidence: 99%

Neuromedin S Regulates Steroidogenesis through Maintaining Mitochondrial Morphology and Function via NMUR2 in Goat Ovarian Granulosa Cells

Sun

Cheng

Wang

et al. 2022

IJMS

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“…Ovaries and uterus, as essential reproductive organs, undergo the development and ovulation of dominant follicles on the ovaries, fertilization in the oviduct, and subsequent development in the uterus, leading to parturition under the influence of reproductive hormones. Transcriptomic studies related to ovaries and follicles have been applied to various species, such as humans ( 30 ), cows ( 17 , 31 , 32 ), pigs ( 33 , 34 ), sheep ( 35 ), chickens ( 36 ), and geese ( 37 ). However, research on the ovaries of equine animals, especially horses, is relatively scarce.…”

Section: Discussionmentioning

confidence: 99%

Differential age-related transcriptomic analysis of ovarian granulosa cells in Kazakh horses

Ren,

Wang,

Zeng

et al. 2024

Front. Endocrinol.

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IntroductionThe Kazakh horse, renowned for its excellence as a breed, exhibits distinctive reproductive traits characterized by early maturity and seasonal estrus. While normal reproductive function is crucial for ensuring the breeding and expansion of the Kazakh horse population, a noteworthy decline in reproductive capabilities is observed after reaching 14 years of age.MethodsIn this study, ovarian granulosa cells (GCs) were meticulously collected from Kazakh horses aged 1, 2, 7, and above 15 years old (excluding 15 years old) for whole transcriptome sequencing.ResultsThe analysis identified and selected differentially expressed mRNAs, lncRNAs, miRNAs, and circRNAs for each age group, followed by a thorough examination through GO enrichment analysis. The study uncovered significant variations in the expression profiles of mRNAs, lncRNAs, miRNAs, and circRNAs within GCs at different stages of maturity. Notably, eca-miR-486-3p and miR-486-y exhibited the highest degree of connectivity. Subsequent GO, KEGG, PPI, and ceRNA network analyses elucidated that the differentially expressed target genes actively participate in signaling pathways associated with cell proliferation, apoptosis, and hormonal regulation. These pathways include but are not limited to the MAPK signaling pathway, Hippo signaling pathway, Wnt signaling pathway, Calcium signaling pathway, Aldosterone synthesis and secretion, Cellular senescence, and NF-kappa B signaling pathway—essentially encompassing signal transduction pathways crucial to reproductive processes.DiscussionThis research significantly contributes to unraveling the molecular mechanisms governing follicular development in Kazakh horses. It establishes and preliminarily validates a differential regulatory network involving lncRNA-miRNA-mRNA, intricately associated with processes such as cell proliferation, differentiation, and apoptosis and integral to the developmental intricacies of stromal follicles. The findings of this study provide a solid theoretical foundation for delving deeper into the realm of reproductive aging in Kazakh mares, presenting itself as a pivotal regulatory pathway in the context of horse ovarian development.

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“…Our protocol used growth factors associated with female gonadal development and GC differentiation: BMP4 and bFGF induces mesoderm commitment which is critical during the development of the urogenital system, while EGF and bFGF is known to aid proliferation of primary GCs in culture (Bruckova et al, 2011; Evseenko et al, 2010; Lawson et al, 1999). Inhibin-alpha and inhibin-beta which are secreted by GCs play a vital role in the maintenance of ovarian follicular development and maturation (Bao et al, 2021; Myers et al, 2009). The final step in our differentiation protocol involved addition to the medium of oocyte-secreted factors in combinations of either a) TGF-α and TGF-β or b) BMP15 and GDF9 – resulting in the GLC-A and GLC-B clones, respectively.…”

Section: Discussionmentioning

confidence: 99%

Generation of a human ovarian granulosa cell model from induced pluripotent stem cells

Hart

Gutiérrez

Biason‐Lauber

2022

Preprint

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SUMMARYSex development is an intricate and crucial process in all vertebrates that ensures the continued propagation of genetic diversity within a species, and ultimately their survival. Perturbations in this process can manifest as variations/differences of sex development (VSD/DSD). Primary gonadal somatic cells – the ovarian granulosa cells (GCs) in the case of women – represent the absolute model to investigate mechanism of disease in VSDs. Collection of these cells in humans is laborious and invasive, while classical animal models fail to recapitulate the human phenotype and function. Furthermore, in patients with the most severe forms of VSD gonadal cells are totally absent. It is therefore vital to develop an alternative cell-model. In view of this, we established an efficient method to reprogram donor-derived urinary progenitor cells (UPs) and differentiate iPSCs into granulosa-like cells (GLCs). The UPs presented a less invasive and high-quality cell source, improving the clinical applicability of the model along with utilising a non-integrative reprogramming method that eliminates alteration of the original genome. This novel GLC model closely resembles human GCs in morphology and marker gene expression of GC cell-fate and essential function. These results provide the prospect to generate patient-specific personalised GC models to investigate mechanism of disease in VSDs and could improve understanding of the intricacies in female gonadal development.

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INHBA transfection regulates proliferation, apoptosis and hormone synthesis in sheep granulosa cells

Cited by 22 publications

References 41 publications

Neuromedin S Regulates Steroidogenesis through Maintaining Mitochondrial Morphology and Function via NMUR2 in Goat Ovarian Granulosa Cells

Neuromedin S Regulates Steroidogenesis through Maintaining Mitochondrial Morphology and Function via NMUR2 in Goat Ovarian Granulosa Cells

Differential age-related transcriptomic analysis of ovarian granulosa cells in Kazakh horses

Generation of a human ovarian granulosa cell model from induced pluripotent stem cells

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