Involvement of Phosphorylated Akt and FOXO3a in the Effects of Growth and Differentiation Factor-9 (GDF-9) on Inhibition of Follicular Apoptosis and Induction of Granulosa Cell Proliferation After In Vitro Culture of Sheep Ovarian Tissue

Monte, Alane Pains Oliveira do; Bezerra, M. É. S.; Menezes, Vanúzia Gonçalves; Gouveia, B.B.; Barberino, Ricássio de Sousa; Lins, T.L.B.G.; Barros, Vanessa Raquel Pinto de; Santos, J.M.S.; Donfack, Nathalie Jiatsa; Matos, Maria Helena Tavares de

doi:10.1007/s43032-020-00409-x

Cited by 9 publications

(4 citation statements)

References 52 publications

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“…Our results also showed that follicular activation has been inhibited by LY294002 in both α-MEM + and 100 pg/ml melatonin treatments. These data suggest that the PI3K pathway could be potentially involved in ovine primordial follicle activation in vitro, which agrees with previous studies in different species (rat: Keating et al, 2009;mouse: Q. Zhao et al, 2014;cat: Thuwanut et al, 2017;sheep: Lins et al, 2021;Monte et al, 2021). The possible explanation for the involvement of the PI3K pathway in follicular activation observed in the α-MEM + is the presence of insulin in the medium, which acts by activating this pathway (Khorami et al, 2015).…”

Section: Discussionsupporting

confidence: 92%

Immunolocalization of melatonin receptor type 1 in the sheep ovary and involvement of the PI3K/Akt/FOXO3a signaling pathway in the effects of melatonin on survival and in vitro activation of primordial follicles

Barberino

Macedo

Lins

et al. 2022

Molecular Reproduction Devel

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This study characterized the expression of melatonin receptor type 1 (MT 1 ) protein in sheep ovaries, evaluated melatonin effects on primordial follicle survival and development after in vitro culture of ovarian tissue and verified the possible involvement of the phosphatidylinositol-3-kinase/protein kinase B/forkhead box O3a (PI3K/Akt/FOXO3a) pathway in the melatonin actions. Ovine ovarian fragments were cultured in α-modified minimum essential medium alone (α-MEM + ) or supplemented with 100, 500, or 1000 pg/ml melatonin for 7 days. PI3K inhibition was performed through pretreatment of ovarian fragments with LY294002. Thereafter, immunohistochemistry was performed to evaluate the expression of cleaved caspase-3, Akt, phosphorylated-Akt, and phosphorylated-FOXO3a (p-FOXO3a). The immunohistochemical localization of the MT 1 receptor protein was documented in sheep preantral and antral follicles. After in vitro culture, 100 pg/ml melatonin showed higher follicular survival and activation than α-MEM + and other melatonin concentrations.After PI3K inhibition, there was an increase in cleaved caspase-3-positive follicles, and a decrease in the primordial follicle activation, Akt phosphorylation, and nuclear exclusion of p-FOXO3a. In conclusion, MT 1 receptor protein is present in the sheep ovary.Furthermore, 100 pg/ml melatonin maintains survival and stimulates activation of primordial follicles through the PI3K/Akt/FOXO3a signaling pathway after in vitro culture of sheep ovarian tissue. K E Y W O R D S hormone receptor, immunohistochemistry, in vitro culture, ovarian cortex, preantral follicle 1 | INTRODUCTION In vitro activation of primordial follicles cultured within the ovary itself is an essential tool to study the early follicle development and optimize the use of the female germ cell pool (Telfer & Zelinski, 2013; J. J. Wang et al., 2017). Studies on in vitro activation of primordial follicles, and subsequent growth and maturation have gained a lot of attention as a potential source of mature eggs for in vitro fertilization and embryo transfer, followed by the delivery of healthy born

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Section: Discussionsupporting

confidence: 92%

Immunolocalization of melatonin receptor type 1 in the sheep ovary and involvement of the PI3K/Akt/FOXO3a signaling pathway in the effects of melatonin on survival and in vitro activation of primordial follicles

Barberino

Macedo

Lins

et al. 2022

Molecular Reproduction Devel

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“…Subsequently, AKT further activates transcription factors such as FRK (forkhead family transcription factor), BAD, and GSK-3, which affect GC proliferation [58]. The level of AKT protein phosphorylation increases following the activation of the PI3K-AKT signaling pathway, which in turn promotes granulosa cell proliferation [59]. To verify the role of NTRK2 in the PI3K-AKT signaling pathway, the protein levels of PI3K and AKT as well as changes in their phosphorylation levels were evaluated in this study.…”

Section: Discussionmentioning

confidence: 99%

NTRK2 Promotes Sheep Granulosa Cells Proliferation and Reproductive Hormone Secretion and Activates the PI3K/AKT Pathway

Jia,

Liu,

Wang

et al. 2024

Animals

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Neurotrophin receptor B (NTRK2), also named TRKB, belongs to the neurotrophic factor family. Previous studies have shown that NTRK2 is associated with high fertility in mammals. However, the molecular mechanism and regulatory pathway of this neurotrophic factor remain unclear. In this study, NTRK2 overexpression and NTRK2-siRNA were constructed to detect the effects of NTRK2 on the proliferation and hormone secretion of the ovarian granulosa cells (GCs) of sheep. We successfully isolated follicular phase granulosa cells in vitro from the ovaries of sheep in simultaneous estrus, and the immunofluorescence results confirmed that NTRK2 was expressed in the collected cells. Subsequently, the effect of NTRK2 on the proliferation of sheep granulosa cells was examined via cell transfection experiments. The results showed that the expression of CDK4 and CyclinD2 was significantly increased after NTRK2 overexpression, while the opposite trend was observed after the inhibition of NTRK2 expression (p < 0.05). The EdU and CCK-8 assays showed that the proliferation rate of sheep GCs was significantly increased after NTRK2 overexpression, while the opposite trend was observed after the inhibition of NTRK2 expression (p < 0.05). Moreover, NTRK2 significantly increased the expression of steroidogenesis-related genes, including steroidogenic acute regulatory protein (STAR) and hydroxy-δ-5-steroid dehydrogenase (HSD3B1), and cytochrome P450 family 19 subfamily A member 1 (CYP19A1). The ELISA results showed that the secretion levels of E2 and P4 significantly increased after NTRK2 overexpression, while the opposite trend was observed after the inhibition of NTRK2 expression (p < 0.05). Previous studies had confirmed that NTRK2 gene belongs to the PI3K-AKT signaling pathway and participates in the signaling of this pathway. This was demonstrated by protein‒protein interaction analysis and NTRK2 belongs to the PI3K-AKT pathway. The modification of PI3K and AKT, markers of the PI3K-AKT pathway, via phosphorylation was increased after NTRK2 overexpression in the sheep GCs, while the opposite trend was observed after the inhibition of NTRK2 expression (p < 0.05). Overall, these results suggest that the NTRK2 gene regulates the proliferation of GCs and the secretion of steroid hormones in sheep, and that it influences the phosphorylation level of the PI3K/AKT signaling pathway. These findings provided a theoretical basis and new perspectives for exploring the regulation of NTRK2 gene in the development of ovine follicles.

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“…Indeed, the concept of in vitro activation (125) has been introduced recently as a clinical treatment in conjunction with autologous ovarian tissue transplantation as a means to restore fertility in patients with premature ovarian insufficiency (126)(127)(128)(129). The in vitro activation effect is mediated via manipulation of the Hippo-PI3K/AKT pathway and is supported by evidence from numerous studies of animal and human ovaries in vivo and in vitro (for example, [130][131][132][133][134][135][136]. Although this research is encouraging, mouse studies have raised concerns that interventions of this nature may lead to the desynchronization of granulosaoocyte interactions, resulting in accelerated follicle depletion in vivo, an effect that may yet prove to negatively impact follicles grown in vitro (137).…”

Section: Novel Tools and Culture Approachesmentioning

confidence: 99%

Therapeutic Potential of In Vitro–Derived Oocytes for the Restoration and Treatment of Female Fertility

Picton

2022

Annu. Rev. Anim. Biosci.

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Considerable progress has been made with the development of culture systems for the in vitro growth and maturation (IVGM) of oocytes from the earliest-staged primordial follicles and from the more advanced secondary follicles in rodents, ruminants, nonhuman primates, and humans. Successful oocyte production in vitro depends on the development of a dynamic culture strategy that replicates the follicular microenvironment required for oocyte activation and to support oocyte growth and maturation in vivo while enabling the coordinated and timely acquisition of oocyte developmental competence. Significant heterogeneity exists between the culture protocols used for different stages of follicle development and for different species. To date, the fertile potential of IVGM oocytes derived from primordial follicles has been realized only in mice. Although many technical challenges remain, significant advances have been made, and there is an increasing consensus that complete IVGM will require a dynamic, multiphase culture approach. The production of healthy offspring from in vitro–produced oocytes in a secondary large animal species is a vital next step before IVGM can be tested for therapeutic use in humans. Expected final online publication date for the Annual Review of Animal Biosciences, Volume 10 is February 2022. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

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Involvement of Phosphorylated Akt and FOXO3a in the Effects of Growth and Differentiation Factor-9 (GDF-9) on Inhibition of Follicular Apoptosis and Induction of Granulosa Cell Proliferation After In Vitro Culture of Sheep Ovarian Tissue

Cited by 9 publications

References 52 publications

Immunolocalization of melatonin receptor type 1 in the sheep ovary and involvement of the PI3K/Akt/FOXO3a signaling pathway in the effects of melatonin on survival and in vitro activation of primordial follicles

Immunolocalization of melatonin receptor type 1 in the sheep ovary and involvement of the PI3K/Akt/FOXO3a signaling pathway in the effects of melatonin on survival and in vitro activation of primordial follicles

NTRK2 Promotes Sheep Granulosa Cells Proliferation and Reproductive Hormone Secretion and Activates the PI3K/AKT Pathway

Therapeutic Potential of In Vitro–Derived Oocytes for the Restoration and Treatment of Female Fertility

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