Involvement of insulin in early development of mouse one-cell stage embryos.
Recent studies have suggested that growth factors and hormones play important roles in cell proliferation and differentiation during early embryonic development. In the present study, we examined the expression and localization of insulin in the mouse oocytes and one-cell stage embryos by quantitative ELISA, RT-PCR, Western blot and immunofluorescence. In the mouse oocytes and one-cell stage embryos, expression of insulin was uniformly distributed in the cytoplasm. We also examined the expression, activity and localization of mTOR (mammalian target of rapamycin) and p70S6K. The expression of mTOR and p70S6K was not significantly different at the cell cycle of mouse one-cell stage embryos. mTOR and S6K were distributed evenly in the cytoplasm at G1, G2 and M phase phase, but at S phase, the distribution of mTOR and S6K was around the pronucleus. At different phases, the activity of mTOR fluctuated. We also used the PI3K specific inhibitor-Wortmannin to investigate the cleavage rate of eggs. The result showed that the rate obviously decreased. When the mTOR specific inhibitor Rapamycin was used, the first mitotic division of the mouse one-cell stage embryo was delayed. These results suggested that insulin was expressed both in mouse oocytes and one-cell stage embryos, and may play functional roles in regulation of mouse early embryogenesis by activating the signal pathway of PI3K/PKB/mTOR/S6K.
Yu
BingZhi
B
Department of Biochemical and Molecular Biology, China Medical University, Shenyang, 110001, China. ybzbiochem@yeah.net
Zhang
Zhe
Z
Deng
Xin
X
Xu
XiaoYan
X
Feng
Chen
C
Li
YanXiao
Y
Cui
Cheng
C
Su
WenHui
W
Zhao
HongMei
H
Yu
DaHai
D
D000730:Androstadienes / Q000378:metabolism; D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle / Q000502:physiology; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology*; D005260:Female; D007328:Insulin / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D009865:Oocytes / Q000166:cytology / Q000502:physiology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics / Q000378:metabolism; D011247:Pregnancy; D047428:Protein Kinase Inhibitors / Q000378:metabolism; D038762:Ribosomal Protein S6 Kinases, 70-kDa / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D020123:Sirolimus / Q000378:metabolism; D058570:TOR Serine-Threonine Kinases; D000077191:Wortmannin; D015053:Zygote / Q000166:cytology / Q000502:physiology*
D000730:Androstadienes; D000903:Antibiotics, Antineoplastic; D002352:Carrier Proteins; D007328:Insulin; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D017853:Phosphotransferases (Alcohol Group Acceptor); D047428:Protein Kinase Inhibitors; D038762:Ribosomal Protein S6 Kinases, 70-kDa; D020123:Sirolimus; D058570:TOR Serine-Threonine Kinases; D000077191:Wortmannin; C546843:mTOR protein, mouse