Objective: To investigate the mechanisms of nitric oxide (NO) in the develo
pment and apoptosis of preimplantation mouse embryos.
Design: Prospective, controlled study.
Setting: Medical college laboratory.
Subject(s): Two-cell embryos from outbred ICR mice.
Intervention(s): Hyperstimulation protocol, two-cell embryos were collected
, then treated with or without an NO synthase inhibitor (L-NAME) or an NO d
onor (SNP) and combined with a cGMP analogue (8-Br-cGMP) or a selective inh
ibitor of NO-sensitive soluble guanylyl cyclase (ODQ).
Main Outcome Measure(s): The development of ICR mouse embryo from two cells
to blastocyst stages in vitro.
Result(s): The development of blastocyst was inhibited by L-NAME in a conce
ntration-dependent manner (0.1-10 muM) and 0.1 muM SNP reversed this effect
(80.5% of control). Annexin-V/propidium iodide and terminal deoxynucleotid
yl transferase-mediated dUTP nick end-labeling techniques demonstrated that
excessive NO (greater than or equal to 10 muM) might induce apoptosis in t
he mouse embryos. 8-Br-cGMP reversed the inhibitory effect of L-NAME and re
scued the embryo growth. ODQ inhibited the embryo development in a dose-res
ponsive fashion (0.1-100 muM) but had no effect in the NO-induced embryo ap
optosis. P53 and Bax were found to be up-regulated during the embryo fragme
ntation.
Conclusion(s): These results indicate that the cGMP pathway might be involv
ed in the NO-regulated embryonic development, but not in NO-induced apoptos
is, for which P53/Bax pathway might be involved. (Fertil Steril(R) 2001,75:
1163-71. (C) 2001 by American Society for Reproductive Medicine.).