Phospholipase C inhibits apoptosis of porcine oocytes cultured in vitro

Abstract

Oocyte apoptosis can be used as an indicator of oocyte quality and development competency. Phospholipase C (PLC) is a critical enzyme that participates in phosphoinositide metabolic regulation and performs many functions, including the regulation of reproduction. In this study, we aimed to explore whether PLC participates in the regulation of apoptosis in porcine oocytes and investigated its possible mechanism. In porcine oocytes, 0.5 μM U73122 (the PLC inhibitor) was considered to be the best concentration to facilitate maturation, and 0.5 μM m-3M3FBS (the PLC activator) was regarded as the most appropriate concentration to inhibit maturation. The percentage of cleavage and blastocysts treated with 0.5 μM U73122 was lower than that of the control group. Furthermore, the percentage of cleavage and blastocysts treated with 0.5 μM m-3M3FBS was higher than that of the control group. The relative PLC messenger RNA (mRNA) expression tested by a quantitative real-time polymerase chain reaction was found to be inhibited by 0.5 μM U73122 or activated by 0.5 μM m-3M3FBS. The relative mRNA abundance of BAK, BAX, CASP3, CASP8, and TP53 and protein abundance of Bak, cleaved caspase-3, caspase-8, and P53 was activated by U73122 or inhibited by m-3M3FBS, while the relative mRNA and protein level of BCL6 showed the opposite trend. The intracellular Ca 2+ concentration increased and the expression of PLCB1 protein also increased in porcine oocytes when they were cultured with 0.5 μM m-3M3FBS for 44 hours. The abundance of proteins PKCβ and CAMKIIα and the expression of several downstream genes (CDC42, NFATc1, NFATc2, NFκB, and NLK) were activated by m-3M3FBS or inhibited by U73122. Our findings indicate that PLC inhibits apoptosis and maturation in porcine oocytes. The intracellular Ca 2+ concentration, two Ca 2+ -sensitive proteins, and several downstream genes were positively regulated by PLC. K E Y W O R D Sapoptosis, Ca 2+ -sensitive proteins, intracellular Ca 2+ concentration, oocytes, phospholipase C, porcine

[Retracted] TRPM7 Upregulate the Activity of SMAD1 through PLC Signaling Way to Promote Osteogenesis of hBMSCs

“…Naltriben, a TRPM7-specific agonist, was used to activate the expression of TRPM7 [1,37,38]. U73122 or M-3M3FBS was reported to inhibit and induce the mRNA expression [39,40] and protein expression [41] of PLC.…”

Protodioscin protects porcine oocytes against H2O2-induced oxidative stress during in vitro maturation

“…Apoptosis is a process of programmed cell death that occurs regularly to ensure a homeostatic balance between the rates of cell formation and cell death, and involves many genes. However, excessive apoptosis can induce degeneration of oocytes and death of early embryos, and also affect normal blastocyst formation [ 24 ]. In this study, the average percentage of apoptotic cells in blastocysts was lower in the 10PD group than in the normal and control groups; however, this difference was not significant.…”

Strengthening the mechanical properties of polysulfide adhesives by introducing polyurethane

“…The viscosity of the prepolymer was the critical factor affecting the practical application. 26 The prepolymer compositions were listed in Table 1, and their corresponding viscosity values were determined by the cone plate viscometer at 60 C. In general, the molar feeding ratio of diisocyanate (-NCO) to liquid polysulfide (-SH) was closely related to the viscosity of polysulfide adhesives. 27 As shown in Figure 2, the viscosity value of prepolymers weakened with a rising R value.…”