PS48 promotes in vitro maturation and developmental competence of porcine oocytes through activating PI3K/Akt signalling pathway

Abstract

The quality of oocytes is closely associated with the successful production of porcine oocytes in vitro. However, the regulating mechanisms of oocytes in vitro maturation (IVM) need to be further improved, and the quality of oocyte maturation in vitro is still inferior to that of oocyte maturation in vivo. Compared with oocytes matured in vivo, the oocytes matured in vitro tend to have such problems as more high precision fertilization rate, low blastocyst formation rate, not fully unified oocyte cytoplasm and nuclei of mature in the current maturation systems (Jiao et al., 2020). Moreover, the mature quality of porcine oocytes has an essential effect on embryo development after in vitro fertilization (IVF), nuclear transfer (NT) and parthenogenetic activation (PA) (Kim et al., 2010). Oocyte

The Role of MAPK3/1 and AKT in the Acquisition of High Meiotic and Developmental Competence of Porcine Oocytes Cultured In Vitro in FLI Medium

“…The addition of the PDK1/PI3K/AKT activator PS48 into the culture medium increased the expansion of cumulus cells and the maturation rate of porcine oocytes [57]. Vice versa, PI3K/AKT inhibitors LY294002 and SH6 reduced the cumulus expansion and developmental competence of porcine and bovine oocytes assessed by blastocyst rate after parthenogenetic activation [38,58,59]. These phenotype changes were accompanied by substantial changes in the expression of specific genes in CCs as well as in mRNA abundance in the oocytes.…”

RNA-Seq Reveals the Underlying Molecular Mechanism of First Cleavage Time Affecting Porcine Embryo Development

“…Thirty oocytes were washed three times with electrical activation medium containing 0.3 mM mannitol, 0.1 mM CaCl 2 , 0.1 mM MgSO 4 , 0.5 mM HEPES, and 0.01 g/L BSA and then transferred into a BTX (BTX, San Diego, CA, USA) covered with electrical activation medium. Electrical stimulation was performed referring to a previous study [ 23 ]. Three direct current (DC) pulses of 50 V/cm for 80 µsec were applied to oocytes with a Cell Fusion-Active device (BLS CF-150/B).…”

“…Blastocysts were fixed with 4% paraformaldehyde in a DPBS (Dulbecco’s Phosphate Buffered Saline) for 10 min at room temperature. After fixation, samples were washed with PBS containing 1 mg/mL polyvinyl alcohol (PBS-PVA) and stained with 10 μg/mL Hoechst 33,342 for 15–20 min [ 23 ]. Thereafter, blastocysts were rinsed with PBS-PVA, mounted on a glass slide, and gently flattened with coverslips.…”

“…cDNA was synthesized by reverse transcription of RNA extracted from early cleavage embryos and late cleavage embryos [ 23 ]. Each 20 μL reaction contained 1 μL cDNA, 1 μL of 10 μmol/L forward and reverse primers, 10 μL FastStart Universal SYBR Green Master (Roche, Basel, Switzerland), and 8 μL nuclease-free water.…”

The influence of ovarian follicular niche on oocyte development

“…The PI3k-Akt pathway is related to follicular quality and oocyte competence in many species such as cattle, swine and humans models, and its alteration is detrimental to oocyte quality (ANDRADE et al, 2017;DU;JIAO et al, 2020). Since both pathways (PI3K and Wnt) relay on the GSK3 enzyme, a multifaceted kinase that plays a critical role in both the canonical pathways in order to control genes related to survival, neurogenesis and follicular development (SHORNING et al, 2020;VALVEZAN;KLEIN, 2012).…”