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In mice deficient in progesterone receptor (PR),
ovulatory-size follicles develop but fail to ovulate,
providing evidence for an essential role for progesterone
and PR in ovulation in mice. However, little is known
about the expression and regulation of PR mRNA in
preovulatory follicles of ruminant species. One objective
of this study was to determine if and when PR mRNA is
expressed in bovine follicular cells during the
periovulatory period. Luteolysis and the LH/FSH surge
were induced with PGF2
and a GnRH
analogue, respectively, and the preovulatory follicle was
obtained at 0, 3.5, 6, 12, 18, or 24 h after GnRH. RNase
protection assays revealed a transient increase in levels
of PR mRNA, that peaked at 6 h after GnRH and declined to
the time 0 value by 12 h, and a second increase at 24 h.
The second objective was to investigate the mechanisms
that regulate PR mRNA expression through in vitro studies
on follicular cells of preovulatory follicles obtained
before the LH/FSH surge. Theca and granulosa cells were
isolated and cultured without or with a luteinizing dose
of LH or FSH, progesterone, LH + progesterone, or LH +
antiprogestin (RU486). Levels of PR mRNA increased in a
time-dependent manner in granulosa cells cultured with LH
or FSH and in theca cells cultured with LH, peaking at 10
h of culture. In contrast, progesterone (200 ng/ml) did
not upregulate mRNA for its own receptor and neither
progesterone nor RU486 affected LH-stimulated PR mRNA
accumulation. Furthermore, RU486 completely blocked
LH-stimulated expression of oxytocin mRNA, indicating that
PR induced by LH in vitro are functional. These results
show that the gonadotropin surge induces a rapid and
transient increase in expression of PR mRNA in both theca
and granulosa cells of bovine periovulatory follicles,
followed by a second rise close to the time of ovulation
and that the first increase in PR mRNA can be mimicked in
vitro by gonadotropins, but not by progesterone. These
results suggest multiple and time-dependent roles for
progesterone and PR in the regulation of periovulatory
events in cattle.
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