Supplement

effect on ovarian as well as adrenal androgen production. These effects may
be, at least in part, related to an improvement of insulin sensitivity and a
decline of insulin level.
Supported by: Intramural Funding.
O-93 Tuesday, October 18, 2016 11:45 AM
GENOME-WIDE DNA METHYLATION CHANGES IN MOUSE
ZYGOTES ASSOCIATED WITH SUPEROVULATION. B. Yu. a,b
a OBGYN, University of Washington, Seattle, WA; b Albert Einstein College
of Medicine, Bronx, NY.
OBJECTIVE: Several animal and human studies have shown Assisted
Reproductive Technologies (ART) procedures can result in abnormal
methylation of some imprinted genes in early embryos. However, it is not
know what genome-wide effect these ART procedures may have, and which
ART manipulations cause these changes. The objective of this study is to
identify DNA methylome changes in pre-implantation mouse embryos after
ovarian hyperstimulation.
DESIGN: Basic research using animal model.
MATERIALS AND METHODS: Using Whole Genome Bisulfite
Sequencing (WGBS) method, we compared DNA methylomes in mouse
pre-implantation embryos from superovulated and negative control
mice (n¼3 in each group). Superovulated mice were injected with 10IU
of PMSG to stimulate multiple oocyte growth, followed by 10IU of
hCG 48 hours later to induce ovulation. The negative control (or natural
mating) group was injected with normal saline at the same time interval
as the study group. Both groups were mated with fertile male mice overnight,
and zygotes were harvested the next morning. All zygotes were
pooled from each female mouse. WGBS was performed according to
our established protocol and libraries were sequenced on HiSeq-2500
with 150-bp single-end reads.
RESULTS: Significant differences in CpG methylation levels were
observed in zygotes from superovulation group compared to natural mating
group, with 31223 differentially methylated regions (600bp in size)
(p