P-81: Distinct Epigenetic Status of Fully Grown and Growing Oocytes Selected by BCB Staining

Authors

  • Forouzanfar M
  • Ghanaei HR
  • Hajian M
  • Hosseini SM
  • Jafarpour F
  • Ostadhosseini S
Abstract:

Background: Since the sheep ‘Dolly’ was successfully cloned by somatic cell nuclear transfer (SCNT), the inefficiency of SCNT cloning and abnormality of cloned animals have been a focus of considerable controversy. Oocyte quality has been shown to contribute to poor somatic cloning and cell reprogramming efficiency with many factors affecting oocyte quality and developmental potential. It has been found that immature oocytes with reduced reproductive performance or slaughtered at the end of their use are heterogeneous in quality. Then selecting immature oocyte is necessary and important for embryos laboratory. It is generally believed that glucose- 6-phosphate dehydrogenase (G6PDH) protein is active in the growing oocyte, but its activity is decreased in oocytes that have finished their growth phase, then likely to have achieved developmental competence. The enzyme G6PDH can degrade brilliant cresyl blue (BCB). Thus, oocytes yielding decreased G6PDH (finished growth phase) show a blue cytoplasm (BCB+) after BCB staining, while growing oocytes (unfinished growth phase) have abundant G6PDH and a colorless cytoplasm (BCB-). Materials and Methods: The oocytes were exposed to 26μM BCB and classified according to their cytoplasm coloration. However, to address the effects of oocyte selection by BCB staining on the nuclear reprogramming capacity of oocytes, embryo quality, we analyze developmental competence, status of epigenetic MII oocyte modifications, embryo quality assessment. Results: Levels of DNA Methylation, H3K9ac and H3K4me3 in BCB + group was significantly higher (89.19±1.49), (90.08±2.18), (98.53±2.62) in compare to the BCB- group (81.95±1.27), (78.73±2.16), (80.32±3.82). The percentage of Parthenogenetic blastocyst development was higher in BCB+ and was significantly different between BCB+ (39.25±9.03) and BCB- (23±7.48) oocytes. Total cell number, TE cell number, ICM cell number: TE ratio of blastocysts were significantly higher in the BCB+ group compared with the BCB- group. Conclusion: Therefore, the BCB test is a useful method for selecting more competent immature sheep oocytes and it made improve Scnt method.

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

P-100: Improved Efficiency of Somatic Cell Nuclear Transfer in Sheep by Persecution of Recipient Oocytes with Brilliant Cresyl Blue Staining

Background: The most important factor affecting the efficiency of somatic cell nuclear transfer (SCNT) is the initial quality of the oocyte. Since the abattoir-derived oocytes are heterogeneous in quality, selection of cohort fully grown/competent oocytes is necessary for proper development of SCNT embryos. Materials and Methods: Glucose-6-phosphate dehydrogenase (G6PDH) is actively expressed i...

full text

Oocytes Selected Using BCB Staining Enhance Nuclear Reprogramming and the In Vivo Development of SCNT Embryos in Cattle

The selection of good quality oocytes is crucial for in vitro fertilization and somatic cloning. Brilliant cresyl blue (BCB) staining has been used for selection of oocytes from several mammalian species. However, the effects of differential oocyte selection by BCB staining on nuclear reprogramming and in vivo development of SCNT embryos are not well understood. Immature compact cumulus-oocyte ...

full text

P-101: Developmental Competence of Bovine Vitrified GV Oocyte Selected by BCB

Background: Cryopreservation of farm animal embryos is nowadays a widely used routine technique. Success in cryopreservation of form animal depends on many factors including: freezing medium, types of cryoproctants, melting process, lipid content in ooplasm and oocyte quality It has been found that immature Oocytes with reduced reproductive performance or slaughtered at the end of their use are...

full text

P-102: Evaluation of G6PDH Enzyme Activity in GV Oocytes Cytoplasm to Recompense The Effects of Vitrification Process

Background: Process of oocyte freezing, seriously damages the molecular structure of the oocyte into the life and impair its ability to develop. It seems Oocytes can partially recompense those damages. The pentose phosphate pathway (PPP) constitutes the major source of NADPH required for the neutralization of reactive oxygen species (ROS). Glucose-6-phosphate dehydrogenase (G6PDH) is the first ...

full text

P-90: Effect of Phosphodiesterase Type3 Inhibitor, Cilostamide, on The Developmental Competence of Ovine OocytesIsolated by Glucose 6-Phosphate Dehydrogenase Activity

Background: The developmental competence of oocytes matured in vitro (IVM) is yet far below than in vivo counterparts. Recent studies suggest that the asynchrony between nuclear/cytoplasmic maturation and the initial low/heterogeneous quality of oocytes are the most important factors affecting IVM success. We investigated whether selection of growing oocytes (based on their glucose 6- phosphate...

full text

P-103: Connection of The G6PDH Activity on Fertilization and Developmental Competency in The Bovine

Background: This study was conducted in order to determine whether BCB staining performed on immature bovine oocytes to assess G6pdhlevel has an impact on ivf/ivp technology in bovine embryos. Both spermatozoa and oocytes metabolize glucose through the pentose phosphate pathway (PPP), and NADPH appears necessary for gamete fusion. Activity of glucose 6-phosphate dehydrogenase (G6PDH), the key e...

full text

My Resources

Save resource for easier access later

Save to my library Already added to my library

{@ msg_add @}


Journal title

volume 7  issue 3

pages  67- 67

publication date 2013-09-01

By following a journal you will be notified via email when a new issue of this journal is published.

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023