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Abstract
Before fertilization, chromatin of both mouse oocytes and
spermatozoa
contain very few acetylated histones. Soon after
fertilization, chromatins of both
gametes become highly acetylated. The same
deacetylation-reacetylation changes
occur with histones of somatic nuclei transferred into
enucleated oocytes. The
significance of these events in somatic chromatin
reprogramming to the totipotent
state is not known. To investigate their importance in
reprogramming, we injected
cumulus cell nuclei into enucleated mouse oocytes and
estimated the histone
deacetylation dynamics with immunocytochemistry. Other
reconstructed oocytes
were cultured before and/or after activation in the
presence of the highly potent
histone deacetylase inhibitor trychostatin A (TSA) for up
to 9 h post-activation. The
potential of TSA-treated and untreated oocytes to develop
to the blastocyst stage and
to full-term was compared. Global deacetylation of
histones in the cumulus nuclei
occurred between 1 to 3 h after injection. TSA inhibition
of histone deacetylation
did not affect the blastocyst rate (37% with and 34%
without TSA treatment), while
extension of the TSA treatment beyond the activation point
significantly increased
the blastocyst rate (up to 81% versus 40% without TSA
treatment) and quality (on
average, 59 vs. 45 cells in day-4 blastocysts with and
without TSA treatment,
respectively). TSA treatment also slightly increased full
term development (from
0.8% to 2.8%). Thus, deacetylation of somatic histones is
not important for
reprogramming, and hyperacetylation might actually improve
reprogramming.
Key words:
Assisted Reproductive Technology •
Developmental biology •
Early development •
Oocyte development
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