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In vitro embryos production after oocytes treatment with forskolin
Daniela Martins Paschoal1, Rosiára Rosária Dias Maziero2, Mateus José Sudano2
1São Paulo State University (UNESP),School of Veterinary Medicine and Animal Science (FMVZ),Department of Animal Reproduction and Veterinary Radiology,Rubião Jr. s/n,Botucatu/SP,Brazil.
Abstract:
The inhibition of nuclear maturation allows time for the oocyte to accumulate molecules that are important for embryonic development. Thus, the objective of this work was to evaluate the effect of blocking oocyte meiosis with the addition of forskolin, an efficient inhibitor of nuclear maturation, in in vitro maturation (IVM) medium. Forskolin was added to the IVM medium for 6 h at concentrations of 0.1 mM, 0.05 mM or 0.025 mM, then the oocytes were allowed to mature in drug-free medium for 18 h. The oocytes were assessed for the stage of nuclear maturation, the activity and distribution of mitochondria, oocyte ultrastructure, the number of viable cells and the apoptosis rate. After forskolin treatment, the oocytes were fertilized in vitro and cultured for 7 days. On day 7, the blastocyst rate, the ultrastructure, the number of intact cells and the apoptosis rate of the blastocysts were measured. No differences were observed for the stage of nuclear maturation of the oocyte, the mitochondrial activity and distribution, the blastocyst rate or total number of intact cells. However, a higher rate of apoptosis was observed in the blastocysts produced from oocytes blocked for 6 h with the higher concentration of forskolin (P < 0.05). We conclude that all the experimental groups reached the MII stage after the addition of forskolin and that the highest concentration of forskolin caused cellular degeneration without harming embryo production on the 7th day.
Insights
Blocking oocyte meiosis with forskolin aids molecule accumulation for embryonic development. While generally safe, high forskolin concentrations increased blastocyst apoptosis, indicating potential cellular degeneration.
Area of Science:
- Reproductive Biology
- Developmental Biology
- Cell Biology
Background:
- Inhibiting nuclear maturation in oocytes allows accumulation of essential molecules for embryonic development.
- In vitro maturation (IVM) is a key technique in assisted reproductive technologies.
Purpose of the Study:
- To evaluate the effect of blocking oocyte meiosis using forskolin on subsequent embryonic development.
- To assess the impact of different forskolin concentrations on oocyte maturation, mitochondrial activity, ultrastructure, and apoptosis.
Main Methods:
- Oocytes were treated with forskolin (0.1 mM, 0.05 mM, 0.025 mM) for 6 hours during IVM.
- Nuclear maturation, mitochondrial activity, ultrastructure, cell viability, and apoptosis were assessed.
- Oocytes were fertilized in vitro, and resulting embryos were cultured for 7 days to evaluate blastocyst formation and quality.
Main Results:
- No significant differences were observed in oocyte nuclear maturation stage, mitochondrial activity, blastocyst rate, or total intact cells across groups.
- A higher apoptosis rate was detected in blastocysts derived from oocytes treated with the highest forskolin concentration (0.1 mM).
- All experimental groups successfully reached the MII stage after forskolin treatment.
Conclusions:
- Forskolin effectively blocks oocyte meiosis, allowing for potential accumulation of developmental molecules.
- While generally not detrimental to embryo production, high forskolin concentrations can induce cellular degeneration and increase blastocyst apoptosis.
