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Promotion of ovarian follicle growth following mTOR activation: synergistic effects of AKT stimulators.

Cheng Y, Kim J, Li XX, Hsueh AJ - PLoS ONE (2015)

Bottom Line: Mature oocytes derived from MHY1485-activated ovarian grafts could be successfully fertilized, leading the delivery of healthy pups.We further treated ovaries with the mTOR activator together with AKT activators (PTEN inhibitor and phosphoinositol-3-kinase stimulator) before grafting and found additive enhancement of follicle growth.Our studies demonstrate the ability of an mTOR activator in promoting follicle growth, leading to a potential strategy to stimulate preantral follicle growth in infertile patients.

View Article: PubMed Central - PubMed

Affiliation: Program of Reproductive and Stem Cell Biology, Department of Ob/Gyn, Stanford University School of Medicine, Stanford, CA, United States of America.

ABSTRACT
Mammalian target of rapamycin (mTOR) is a serine/threonine kinase and mTOR signaling is important in regulating cell growth and proliferation. Recent studies using oocyte- and granulosa cell-specific deletion of mTOR inhibitor genes TSC1 or TSC2 demonstrated the important role of mTOR signaling in the promotion of ovarian follicle development. We now report that treatment of ovaries from juvenile mice with an mTOR activator MHY1485 stimulated mTOR, S6K1 and rpS6 phosphorylation. Culturing ovaries for 4 days with MHY1485 increased ovarian explant weights and follicle development. In vivo studies further demonstrated that pre-incubation of these ovaries with MHY1485 for 2 days, followed by allo-grafting into kidney capsules of adult ovariectomized hosts for 5 days, led to marked increases in graft weights and promotion of follicle development. Mature oocytes derived from MHY1485-activated ovarian grafts could be successfully fertilized, leading the delivery of healthy pups. We further treated ovaries with the mTOR activator together with AKT activators (PTEN inhibitor and phosphoinositol-3-kinase stimulator) before grafting and found additive enhancement of follicle growth. Our studies demonstrate the ability of an mTOR activator in promoting follicle growth, leading to a potential strategy to stimulate preantral follicle growth in infertile patients.

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Treatment with MHY1485 and subsequent grafting allowed the derivation of mature oocytes and healthy offspring.A) Early embryonic development of oocytes after mTOR activator treatment. Ovaries were treated with MHY1485 for 2 days to activate follicles, followed by grafting into hosts for 5 days. Hosts were then treated with eCG and hCG. At 12h after hCG injection, mature oocytes were obtained and fertilized with sperm before culturing for 4 days. B) Percentage of oocytes developed into each embryonic stage. Early embryonic development for mice at 25 days of age served as controls. N = 30. C) Some 2-cell stage embryos were transferred into pseudopregnant hosts and pups were delivered.
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pone.0117769.g003: Treatment with MHY1485 and subsequent grafting allowed the derivation of mature oocytes and healthy offspring.A) Early embryonic development of oocytes after mTOR activator treatment. Ovaries were treated with MHY1485 for 2 days to activate follicles, followed by grafting into hosts for 5 days. Hosts were then treated with eCG and hCG. At 12h after hCG injection, mature oocytes were obtained and fertilized with sperm before culturing for 4 days. B) Percentage of oocytes developed into each embryonic stage. Early embryonic development for mice at 25 days of age served as controls. N = 30. C) Some 2-cell stage embryos were transferred into pseudopregnant hosts and pups were delivered.

Mentions: We further treated day 10 ovaries with MHY1485 for 2 days in vitro, followed by grafting them into adult hosts treated daily with FSH for 5 days. As shown in Fig. 2A, treatment with MHY1485 increased graft weights. Following histological analyses (Fig. 2B) and follicle counting (Fig. 2C), increases in the development of antral/preovulatory follicles were apparent, together with a decrease of early secondary follicles and an increase of primary follicles. Using this model, we further treated the hosts with eCG for 2 days to promote the growth of preovulatory follicles, followed by an injection of hCG to promote oocyte maturation. At 12h after hCG injection, mature oocytes were punctured from preovulatory follicles for in vitro fertilization. As shown in Fig. 3A, oocytes obtained from MHY1485-pretreated ovaries could develop into blastocysts. As compared with mature oocytes obtained from day 25 mice without MHY1485 treatment, which served as controls, comparable early embryonic development was apparent based on the percentage of oocytes developing into each embryonic stage (Fig. 3B). Some of the 2-cell embryos derived from MHY1485-pretreated grafts were transferred into pseudopregnant surrogate mothers and healthy pups were delivered (Fig. 3C).


Promotion of ovarian follicle growth following mTOR activation: synergistic effects of AKT stimulators.

Cheng Y, Kim J, Li XX, Hsueh AJ - PLoS ONE (2015)

Treatment with MHY1485 and subsequent grafting allowed the derivation of mature oocytes and healthy offspring.A) Early embryonic development of oocytes after mTOR activator treatment. Ovaries were treated with MHY1485 for 2 days to activate follicles, followed by grafting into hosts for 5 days. Hosts were then treated with eCG and hCG. At 12h after hCG injection, mature oocytes were obtained and fertilized with sperm before culturing for 4 days. B) Percentage of oocytes developed into each embryonic stage. Early embryonic development for mice at 25 days of age served as controls. N = 30. C) Some 2-cell stage embryos were transferred into pseudopregnant hosts and pups were delivered.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4340052&req=5

pone.0117769.g003: Treatment with MHY1485 and subsequent grafting allowed the derivation of mature oocytes and healthy offspring.A) Early embryonic development of oocytes after mTOR activator treatment. Ovaries were treated with MHY1485 for 2 days to activate follicles, followed by grafting into hosts for 5 days. Hosts were then treated with eCG and hCG. At 12h after hCG injection, mature oocytes were obtained and fertilized with sperm before culturing for 4 days. B) Percentage of oocytes developed into each embryonic stage. Early embryonic development for mice at 25 days of age served as controls. N = 30. C) Some 2-cell stage embryos were transferred into pseudopregnant hosts and pups were delivered.
Mentions: We further treated day 10 ovaries with MHY1485 for 2 days in vitro, followed by grafting them into adult hosts treated daily with FSH for 5 days. As shown in Fig. 2A, treatment with MHY1485 increased graft weights. Following histological analyses (Fig. 2B) and follicle counting (Fig. 2C), increases in the development of antral/preovulatory follicles were apparent, together with a decrease of early secondary follicles and an increase of primary follicles. Using this model, we further treated the hosts with eCG for 2 days to promote the growth of preovulatory follicles, followed by an injection of hCG to promote oocyte maturation. At 12h after hCG injection, mature oocytes were punctured from preovulatory follicles for in vitro fertilization. As shown in Fig. 3A, oocytes obtained from MHY1485-pretreated ovaries could develop into blastocysts. As compared with mature oocytes obtained from day 25 mice without MHY1485 treatment, which served as controls, comparable early embryonic development was apparent based on the percentage of oocytes developing into each embryonic stage (Fig. 3B). Some of the 2-cell embryos derived from MHY1485-pretreated grafts were transferred into pseudopregnant surrogate mothers and healthy pups were delivered (Fig. 3C).

Bottom Line: Mature oocytes derived from MHY1485-activated ovarian grafts could be successfully fertilized, leading the delivery of healthy pups.We further treated ovaries with the mTOR activator together with AKT activators (PTEN inhibitor and phosphoinositol-3-kinase stimulator) before grafting and found additive enhancement of follicle growth.Our studies demonstrate the ability of an mTOR activator in promoting follicle growth, leading to a potential strategy to stimulate preantral follicle growth in infertile patients.

View Article: PubMed Central - PubMed

Affiliation: Program of Reproductive and Stem Cell Biology, Department of Ob/Gyn, Stanford University School of Medicine, Stanford, CA, United States of America.

ABSTRACT
Mammalian target of rapamycin (mTOR) is a serine/threonine kinase and mTOR signaling is important in regulating cell growth and proliferation. Recent studies using oocyte- and granulosa cell-specific deletion of mTOR inhibitor genes TSC1 or TSC2 demonstrated the important role of mTOR signaling in the promotion of ovarian follicle development. We now report that treatment of ovaries from juvenile mice with an mTOR activator MHY1485 stimulated mTOR, S6K1 and rpS6 phosphorylation. Culturing ovaries for 4 days with MHY1485 increased ovarian explant weights and follicle development. In vivo studies further demonstrated that pre-incubation of these ovaries with MHY1485 for 2 days, followed by allo-grafting into kidney capsules of adult ovariectomized hosts for 5 days, led to marked increases in graft weights and promotion of follicle development. Mature oocytes derived from MHY1485-activated ovarian grafts could be successfully fertilized, leading the delivery of healthy pups. We further treated ovaries with the mTOR activator together with AKT activators (PTEN inhibitor and phosphoinositol-3-kinase stimulator) before grafting and found additive enhancement of follicle growth. Our studies demonstrate the ability of an mTOR activator in promoting follicle growth, leading to a potential strategy to stimulate preantral follicle growth in infertile patients.

Show MeSH
Related in: MedlinePlus