Phospholipase C (PLC)는 다양한 세포주에서 세포내 신호전달에 중요한 역할을 한다고 알려져 있으나, 생쥐 난자성숙 과정과 착성전 배아발생 과정에서 PLC의 역할과 발현은 아직 연구된 바 없다. 본 연구에서는 난자성숙과 착상전 배아발생 과정에서 생쥐의 PLC β1과 γ1의 유전자 발현을 조사하기 위하여 한 개의 난자 혹은 배아에서 추출된 total RNA를 사용하여 경쟁적 RT-PCR 방법으로 mRNA를 정량하였다. PL

It has been wel known that phospholipase C(PLC) plays an important role in the intracellular signaling in a variety of cell types. However, involvement of PLC in mouse oocyte maturation and preimplantation embryo development remains unknown. The present study examined the expression patterns of the mouse PLC β1 and γ1 during oocyte maturatio and preimplantation embryo development study examined the expression patterns of the mouse PLC β1 and γ1 during oocyte maturation and preimplantation embryo development by the competitive reverse transcription-polymerase chain reaction (RT-PCR method). PLC γ1 mRNA (0.1 fg) was readily detected in germinal vesicle (GV)-stage oocyte and its level was reduced as meiotic resumption proceeded. PLC-β1 mRNA (<0.1 fg) as detected at low level at GV-stage oocytes and scarcely detected at germinal vescle breakdown (GVBD)-stage oocytes. After fertilization, both PLC β1 and γ1 mRNA levels began to increase at morula-stage embryos (0.2 fg) and were more prominent in blastocyst-stage embryos(1 fg). to elucidate the possible involvement of PLC via protein kinase C(PKC) pathway during oocyte maturation and preimplantation embryo development , the effects of sphingosine (PKC inhibitor), sn-diC8 (PKC activator) anc U73122 (PLC ingibitor) were examined. Treatment of GV-stage oocytes with sphingosine (20 μM) facilitated the meiotic resuption by 10-20 over the control within 1 h as judged by GVBD, whereas U73122 failed to show any significant effect. U73122 (10 μM) effectively blocked the compaction of morula, while sn-diC8 (50 μM). In summary, the present study shows that the mouse PLC β1 and γ1 are expressed in a developmental stage-specific manner and PLC-PKC pathway may be involved in early preimplantation embryo development.

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