R18 Peptide 14-3-3 Inhibitor Review

Results Summary

In a preliminary experiment to examine YWHA protein function, we injected oocytes with the synthetic YWHA-inhibitory peptide R18. For these experiments, we determined the threshold or critical concentration of dbcAMP, just enough to maintain prophase I arrest in at least 75% of oocytes cultured overnight, to be 0.05?mg/mL. Oocyte maturation experiments in this concentration of dbcAMP permitted us to examine treatments that could promote maturation compared to control cells in which meiotic arrest is maintained but is not dominated by the activation of PKA by dbcAMP. Microinjection of 0.5?mg/mL R18 into oocytes, followed by overnight incubation in media containing 0.05?mg/mL dbcAMP, promoted release from meiotic arrest and GVBD in a larger number of cells compared to uninjected control oocytes or control oocytes injected with deionized water. Ten of 15 (67%) of oocytes underwent GVBD following R-18 injection, while only 4 of 16 (25%) did so when injected with deionized water, and only 5 of 18 (28%) control uninjected oocytes underwent GVBD (Fig. 5a). Examination of the data in the R18 study using Fisher’s exact test and pairwise comparison of the percentage of GVBD in the control uninjected cells and control water-injected cells revealed no significant difference between those groups (P =?>?0.999). Pairwise comparison of the R18-injected cells to the control water-injected cells revealed a significant difference (P?