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Experimental Studies |
Department of Obstetrics and Gynecology, University of Illinois College of Medicine, Chicago, Illinois 60612
Address all correspondence and requests for reprints to: Dr. Asgi Fazleabas, Department of Obstetrics and Gynecology, University of Illinois, 820 South Wood Street (M/C 808), Chicago, Illinois 60612-7313.
As estrogen and progesterone are proposed regulators of luteal
function, this study was undertaken to correlate the presence of
receptors for these steroids with luteal function during early
pregnancy. Corpora lutea (CL) were obtained from nonpregnant baboons
during the midluteal [ML; days 7–8 postovulation (PO)] and late
luteal (LL; days 11–12 PO) phases of the menstrual cycle or from
pregnant baboons on days 18, 25, 29, or 31–33 PO. Estrogen and
progestin receptors (ER and PR, respectively) and 3ß-hydroxysteroid
dehydrogenase (3ßHSD) were detected by immunocytochemistry using
specific monoclonal (H222 for ER; JZB39 for PR) or polyclonal (S683 for
3ßHSD) antibodies. In addition, ribonucleic acid (RNA) was extracted
from CL, processed for Northern blot analysis, and probed with
complementary DNAs to human PR, human 3ßHSD, and rat aromatase.
Levels of messenger RNA (mRNA) for 3ßHSD were quantified by laser
densitometric scanning, and the data were normalized to the expression
of a housekeeping gene (glyceraldehyde-3-phosphate dehydrogenase) to
correct for loading differences. CL did not demonstrate specific
nuclear stain for ER at any stage of the menstrual cycle or pregnancy.
In contrast, PR-positive cells were present during the ML phase, but
decreased during the LL phase (P < 0.05).
PR-positive cells were maintained during early pregnancy at levels
comparable to the ML phase (P > 0.05). Staining
for 3ßHSD was present at all stages of the cycle and pregnancy.
Although the percent of 3ßHSD-positive cells appeared to decrease as
pregnancy proceeded, this was not statistically different
(P > 0.05). The complementary DNA to PR hybridized
to multiple transcripts (
4.4, 3.1, 1.6, and 0.95 kilobases) in CL of
the cycle. A single transcript (1.8 kilobases) for 3ßHSD was
present in CL at all stages of the cycle and pregnancy. The level of
3ßHSD mRNA was highest during the ML phase and declined significantly
(P < 0.05) during the LL phase and early
pregnancy. Three transcripts (3.6, 3.0, and 1.7 kilobases) for
aromatase were detected in CL of the cycle and pregnancy. Aromatase
mRNA increased during early pregnancy. These results support the
concept of PR-mediated events, but not ER-regulated processes in the
primate CL. Furthermore, the data suggest that the steroidogenic
enzymes 3ßHSD and aromatase are differentially regulated during early
pregnancy.
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