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and ß in the Baboon Fetal Adrenal Gland1
Departments of Obstetrics, Gynecology, Reproductive Sciences, and Physiology (E.D.A., J.S.B.), Center for Studies in Reproduction, University of Maryland School of Medicine, Baltimore, Maryland 21201; and the Department of Physiological Sciences (W.A.D., M.G.L., G.J.P.), Eastern Virginia Medical School, Norfolk, Virginia 23501
Address all correspondence and requests for reprints to: Eugene D. Albrecht, Ph.D., Department of Obstetrics, Gynecology, and Reproductive Sciences, University of Maryland School of Medicine, Bressler Research Laboratories 11–019, 655 West Baltimore Street, Baltimore, Maryland 21201. E-mail: ealbrech{at}umaryland.edu
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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mRNA levels, determined by competitive RT-PCR, were
approximately 7-fold greater (P < 0.02) in the
fetal adrenal of late (187.8 ± 40.3 attomoles/µg RNA) compared
with mid (27.4 ± 5.4 attomoles/µg RNA) gestation. Moreover,
estrogen receptor mRNA expression, determined by quantitative
in situ hybridization, was approximately 2.5-fold
greater (P < 0.05) in the definitive/transitional
zones (21.6 ± 0.5 silver grains/0.025 mm2) than in
the fetal zone (8.3 ± 1.5 grains/0.025 mm2) late in
gestation. The mRNA for the ß-isoform of the estrogen receptor was
also expressed in the baboon fetal adrenal cortex. There was a gradient
of immunocytochemical staining for the estrogen receptor and ß
proteins, with extensive immunoreactivity for both isoforms in the
definitive zone and lower staining in the transitional zone and the
fetal zone. In summary, the results of the present study show that
estrogen receptor and ß were expressed in the fetal and
definitive/transitional zones of the baboon fetal adrenal cortex at mid
and late gestation. The presence of the estrogen receptor provides a
mechanism for mediating the action of estrogen in modulating
ACTH-dependent and cortical zone-specific development and function of
the primate fetal adrenal gland. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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5-3ß-hydroxy-steroid dehydrogenase (3ßHSD)
enzyme needed for the formation of aldosterone and cortisol,
respectively, hormones required for neonatal maturation and
homeostasis. The regulation of the maturation and function of this
important endocrine gland during fetal and neonatal stages of
development, however, is incompletely understood.
We have demonstrated, as illustrated in Fig. 1
, that estrogen has two different
actions on fetal adrenal development and function in the primate.
Estrogen has an indirect action on fetal adrenal development in the
second half of baboon pregnancy by regulating the placental 11ßHSD-1
and -2 enzyme system and thus the increase in transplacental oxidation
of cortisol to cortisone that results in fetal pituitary ACTH release
and maturation of the fetal adrenal transitional zone for cortisol
synthesis (reviewed in Refs. 1, 3, 4). In contrast, estrogen
elicits a direct action on the baboon fetal adrenal by inhibiting the
responsivity of the fetal zone to ACTH with respect to the production
of DHA (Fig. 1), as shown in vitro (5, 6) and in
vivo (7), possibly to maintain a physiologically normal balance of
estrogen production. These two effects of estrogen on fetal adrenal
function are consistent with the observation that in incubates of
dispersed baboon fetal adrenal cells the formation of cortisol
increased, whereas that of DHA decreased, when expressed on a per cell
basis between mid- and late gestation (8, 9). Therefore, we have
hypothesized that there is an estrogen-dependent divergence in cortical
zone-specific development and function in the fetal adrenal gland with
advancing gestation (4, 10, 11). This regulatory process may involve
the ACTH receptor, because although ACTH receptor messenger RNA (mRNA)
expression was enhanced in the transitional/definitive zones in late
gestation, ACTH receptor expression declined in the fetal zone between
mid- and late gestation (10, 11).
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and ß in
the primate fetal adrenal gland have not been determined. Therefore, a
developmental approach was used in the present study to determine the
potential expression of mRNA and protein for the estrogen receptor by
RT-PCR, in situ hybridization, and immunocytochemistry in
the fetal and definitive/transitional zones of the baboon fetal adrenal
gland at mid- and late gestation. |
Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Competitive RT-PCR
RNA preparation. The mRNA levels for the estrogen
receptor were quantified by the competitive RT-PCR assay
established by Riedy et al. (14) as modified in our
laboratory (15). Total RNA was obtained from baboon fetal adrenals by 4
M guanidine isothiocyanate homogenization,
chloroform-isoamyl alcohol extraction, and cesium chloride
centrifugation.
Primer sequence. The estrogen receptor oligonucleotide
primers were synthesized by Life Technologies, Inc. (Grand
Island, NY); they were selected from the human estrogen receptor
complementary DNA (cDNA) sequence (16) and flanked a portion of the
sequence spanning exons 4–7 and overlapping introns D, E, and F. The
primers were as follows: primer 1: downstream, 5'-TTC TCT TCC AGA GAC
TTC AGG GTG CTC ATG CGG AAC CGA GAT GAT-3' (position 1642–1623 linked
to 1544–1525); primer 2: upstream, 5'-AAT TTA ATA CGA CTC ACT
ATA GGG AGA TCC TAC CAG ACC CTT CAG-3' (position T7 polymerase
sequence; the underlined sequences are linked to
1226–1245); primer 3: downstream, 5'-TTC CAG AGA CTT CAG GGT GC-3'
(position 1642–1623); and primer 4: upstream, 5'-GAT CCT ACC AGA CCC
TTC AG-3' (position 1226–1245).
Construction of internal standard RNA. The estrogen
receptor competitive reference standard (CRS) was prepared by the
method of Riedy et al. (14), using RT-PCR to generate the
cDNA template and transcription with T7 polymerase. Total RNA (2 µg)
from baboon fetal adrenals was reversed transcribed at 42 C for 60 min
in a reaction mixture (20 µl) containing 1 mM each of
deoxy (d)-ATP, dCTP, dGTP, and dTTP (Promega Corp.,
Madison, WI); 1 mM dithiothreitol; 200 U SUPERSCRIPT
ribonuclease (RNase) H-RT (Life Technologies, Inc.); 40 U
RNAguard (Pharmacia Biotech, Piscataway, NJ); 50
mM Tris-HCl; 75 mM KCl; 3.0 mM
MgCl2; and 250 ng random primers (Life Technologies, Inc.). After 60 min, the RT mixture was incubated at 70 C for 15
min and cooled to 4 C, and 5 µl were added to a PCR mixture (45 µl)
containing 0.2 mM each of dATP, dCTP, dGTP, and dTTP; 10
mM Tris-HCl; 1.5 mM MgCl2; 50
mM KCl; 1.25 U cloned thermus aquaticus DNA polymerase
(Amplitag, Perkin Elmer Corp./Cetus, Norwalk, CT); and 20
pmol each of primers 1 and 2. PCR was performed in a programmable
thermal cycler (MJ Research, Inc., Cambridge, MA), and the
sample was amplified in 28 or 34 sequential cycles at 94 C for 1 min,
60 C for 1 min, and 72 C for 2 min. After the last cycle, the sample
was incubated for an additional 5 min at 72 C. An aliquot of the PCR
reaction was fractionated by electrophoresis in a 2% agarose gel and
visualized in ethidium bromide. The 417-bp amplified target
(i.e. wild-type) mRNA strand and 339-bp CRS synthesized with
the MEGAscript T7 in vitro transcription kit (Ambion, Inc., Austin, TX) were gel purified using the QIAEX II gel
extraction kit (QIAGEN, Valencia, CA).
Competitive RT-PCR. A constant amount of fetal adrenal total
RNA (3.0 µg) was added to the RT mixture containing 3-fold serial
dilutions of the estrogen receptor -CRS (30–2438 attomoles). After
completion of the RT, 20 pmol each of primers 3 and 4 were added for
the PCR. Negative controls in which either RNA or RT was omitted from
the reaction were also performed. The PCR products were fractionated by
electrophoresis, visualized with a UV transilluminator, and
photographed using type 665 positive/negative film (Polaroid Corp.,
Cambridge, MA).
Quantification of estrogen receptor . Photographs
(negative image), representing the amplified products, were analyzed by
audioradiography using a model 620 video densitometer (Bio-Rad Laboratories, Inc., Richmond, CA). The intensity of the
amplified products was represented as the relative area under each
sample band. A correction factor (17) was used to account for the
differences in size of the target and CRS cDNAs. The logarithm (log) of
the ratio of CRS to target area was plotted as a function of the
concentration of estrogen receptor -CRS added to each PCR reaction.
The concentration of estrogen receptor mRNA was determined where
the ratio of CRS/target area was equal to 1 (i.e.
equivalence point).
In situ hybridization
The methods for in situ hybridization previously
employed in our laboratories (18) were modified essentially as
described by Liuzzi et al. (19) for localization and
zone-specific expression of estrogen receptor and ß mRNA in the
baboon fetal adrenal gland at mid- and late gestation. Sense and
antisense estrogen receptor and ß riboprobes were prepared from
the SacII-SmaI fragment (bases 222–478) of the
human estrogen receptor (IGBMC, Strassburg, France) and the
SacII-PstI fragment (bases 1–308) of the human
estrogen receptor ß (provided by P. Webb, University of California,
San Francisco, CA) cDNAs, which were subcloned into pBluescript SK
(Stratagene, La Jolla, CA), and orientations were verified
by DNA sequencing (20). Probes were radiolabeled with
[33P]UTP using a Riboprobe In Vitro
Transcription Kit (Promega Corp.), and unincorporated
isotope was removed by filtration through a TE Midi Select Column (5
Prime-3 Prime, Boulder, CO).
At the time of hybridization, paraffin-embedded fetal adrenal sections
(4 µm) were incubated (10 min) with 0.02 M HCl, washed,
incubated (70 C, 30 min) with 2 x SSC (1 x SSC = 0.15
M NaCl and 0.015 M sodium citrate buffer, pH
7.2), and then treated (15 min; 37 C) with 5 µg/ml proteinase K
(Sigma Chemical Co., St. Louis, MO). After incubation (10
min) with 10% buffered formalin, sections were rinsed in PBS and
treated with 0.25% acetic anhydride in 0.1 M
triethanolamine hydrochloride-0.9% NaCl. After dehydration in ethanol,
delipidation in chloroform, and rehydration, sections were hybridized
overnight at 55 C with labeled sense or antisense estrogen receptor
or ß riboprobe (106 dpm/slide) in 50 µl hybridization
buffer consisting of 10 ml formamide, 1 ml 1 M Tris (pH
8.0), 0.1 ml 0.5 M EDTA, 500 µl yeast transfer RNA
(Sigma Chemical Co.), 400 µl 50 x Denhardt’s
solution (Sigma Chemical Co.), 20 µl polyadenylic acid
(0.1 µg/µl), 20 µl salmon sperm DNA (500 µg/ml; Life Technologies, Inc., Grand Island, NY), 2 g dextran sulfate,
0.8 ml 5 M NaCl, and diethylpyrocarbonate-H2O.
After incubation, sections were washed repeatedly in 4 x SSC,
serially dehydrated, and washed in 50% formamide, 300 nM
NaCl, 25 mM Tris, and 1 mM EDTA. After a 10-min
rinse in 2 x SSC, nonspecific hybridized probe was removed by
incubation (10 min; 37 C) with RNase A (26 µg/ml; Promega Corp.) and RNase T1 (2000 U; Promega Corp.) in
RNase buffer consisting of 500 mM NaCl, 10 mM
Tris, and 1 mM EDTA (pH 8.0). Subsequently, slides were
washed twice in RNase buffer at 50 C for 15 min each time and then four
times in 2 x SSC for 15 min each time at 50 C. Slides were then
washed in 0.2 x SSC at 60 C for 2 h before serial
dehydration and vacuum drying. Slides were coated with
Kodak NTB2 nuclear emulsion (Eastman Kodak Co., Rochester, NY) diluted 1:1 with distilled water, and after
21 days were developed and lightly counterstained with hematoxylin.
Specific zonal localization of estrogen receptor expression in the
fetal adrenal was confirmed by both light microscopy and darkfield
optics. Quantification of estrogen receptor mRNA expression was
performed by counting the specific number of silver grains per 0.025
mm2 using an Optiphot 2 microscope attached to a
video-based Image-1 analysis system (Universal Imaging Corp., West
Chester, PA). Spatial calibration of the imaging system using a x40
objective was 512 pixels along the horizontal axis and 480 pixels along
the vertical axis. An average of 10 different 145 x 180-µm
areas of the fetal zone and the definitive/transitional zones were
examined. After establishing a gray area, cells with a grain density
10-fold greater than background were considered estrogen receptor
positive. All silver grain counts (number per 0.025 mm2)
were averaged, and a single value for each animal was determined and
used to calculate the overall group mean value.
Laser capture microdissection
Zone-specific cells of fetal adrenal glands obtained in late
gestation were isolated by laser capture microdissection using an
Arcturus Pix Cell LCM System (Arcturus, Los Angeles. CA) and methods
described by the manufacturer and described originally by Emmert-Buck
et al. (21). Briefly, frozen adrenal tissue was sectioned (4
µm) onto nontreated glass microscope slides, and the
definitive/transitional zone was demarcated from the fetal zone by
previously determined immunocytochemical expression/image analysis of
definitive/transitional zone-specific 3ßHSD. Approximately 200 cells
from each zone were microdissected and transferred to microfuge tubes,
RNA was extracted using the RNeasy kit (QIAGEN), and
estrogen-receptor mRNA was determined by RT-PCR as described above.
As a control, endometrium was obtained by laser capture from a section
of frozen baboon uterus from the late follicular phase of the menstrual
cycle, and estrogen receptor mRNA was determined by RT-PCR
concomitantly with cells from the fetal adrenal.
Immunocytochemistry
Estrogen receptor and ß protein expression in the baboon
fetal adrenal was determined by immunocytochemistry using methods
previously established in our laboratory (22). Fetal adrenals were
sectioned at 4 µm, and tissue was mounted on slides, heat fixed,
placed in a microwave for 20 min, and endogenous peroxidase inhibited
with 0.4% H2O2 in methanol. Tissue was then
incubated overnight at 4 C with polyclonal NCL-ER6 F11 antibody to the
rat estrogen receptor (Vector Laboratories, Inc.,
Burlingame, CA) diluted 1:400 in 5% goat serum or polyclonal PA1–313
antibody to the C-terminal amino acid 467–485 of the human estrogen
receptor ß (Affinity BioReagents, Inc., Golden, CO)
diluted in PBS (2 µg/ml). The expression of 3ßHSD was determined by
incubation of nonmicrowaved adrenal sections overnight at 4 C with
polyclonal antibody to rabbit antihuman 3ßHSD (supplied by Dr. Ian
Mason, University of Edinburgh, Edinburgh, Scotland) diluted 1:2500 in
5% goat serum. Sections were then washed and incubated with
biotinylated goat antimouse IgG (estrogen receptor ) or antirabbit
IgG (estrogen receptor ß and 3ßHSD; Vector Laboratories, Inc.) and then with Vectastain Elite Kit
(Vector Laboratories, Inc.). After rinsing in PBS,
sections were stained with diaminobenzidene
(DAB)-imidazole-H2O2 (estrogen receptor ß and
3ßHSD) or DAB-nickel sulfate (estrogen receptor , 0.250 g/10 ml,
in 0.05 M Tris buffer, pH 7.2; Sigma Chemical Co.) as described by Berghorn et al. (23). Tissue
sections were then counterstained with Gill’s hematoxylin and mounted
in Biomount.
Statistics
mRNA levels were analyzed by one-way ANOVA and Newman-Keuls
multiple comparison test or by Student’s t test for
independent observations to identify significant differences between
individual group means.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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mRNA levels by competitive RT-PCR shows a representative
quantitative analysis of estrogen receptor mRNA levels by
competitive RT-PCR in whole baboon fetal adrenal tissue obtained at
mid- and late gestation. The expected 417-bp estrogen receptor target
product and the 339-bp estrogen receptor CRS product generated by PCR
are shown in Fig. 2A. In contrast, there was no PCR product when either
RNA or RT was omitted from the reaction (data not shown). The slopes of
the log of the CRS to target areas plotted as a function of increasing
amounts of CRS were similar for RNA obtained from the fetal adrenal at
mid- and late gestation (Fig. 2B), indicating no difference in
amplification efficiency. However, analysis of the equivalence points
from each plot indicated that fetal adrenal estrogen receptor mRNA
levels were different at mid- and late gestation. Thus, overall mean
(±SE) mRNA levels for estrogen receptor were
approximately 7-fold greater (P < 0.02) in the fetal
adrenal of late (187.8 ± 40.3 attomoles/µg RNA) compared with
mid (27.4 ± 5.4 attomoles/µg RNA) gestation (Fig. 2C).
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and ß mRNA expression by in situ
hybridization mRNA expression are shown for sections of baboon fetal
adrenals of mid (Fig. 3, A and C) and
late (Fig. 3, B, D, and F) gestation incubated with
33P-labeled antisense (Fig. 3, A, B, and F) or sense (Fig. 3, C and D) probes. The specificity of estrogen receptor labeling was
evident by the relative absence of silver grains over cells with the
sense riboprobe and the selectively greater distribution of silver
grains over cells of the outer definitive/transitional zones (Fig. 3B, DZ) compared with the fetal zone (Fig. 3F, FZ) in fetal adrenals of
late gestation. Estrogen receptor mRNA was expressed in the fetal
adrenal at midgestation (Fig. 3A), a time when the gland is comprised
almost exclusively of the fetal zone. In late gestation there was
extensive expression of estrogen receptor mRNA in the outer
definitive/transitional zones (Fig. 3B), which also stained selectively
and intensely for 3ßHSD (not shown). Estrogen receptor was still
expressed in the fetal zone late in gestation (Fig. 3F), although at an
apparently lower level than in the definitive/transitional zones.
Localization of the estrogen receptor in the endometrium of the
baboon uterus, a well established site of this steroid receptor, is
shown in Fig. 3E. When quantified by image analysis, estrogen receptor
mRNA levels (silver grains per 0.025 mm2) in the fetal
adrenal appeared to decline, although not significantly, in the fetal
zone between mid- and late gestation (Fig. 4) and were approximately 2.5-fold
greater (P < 0.05) in the definitive/transitional
zones (21.6 ± 0.5) than in the fetal zone (8.3 ± 1.5) in
late gestation.
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mRNA observed by in situ
hybridization in the baboon fetal adrenal was confirmed in tissue
isolated by laser capture microdissection and analyzed by RT-PCR (Fig. 5). Thus, estrogen receptor mRNA was
detected in both the fetal (lane 2) and definitive/transitional (lanes
3 and 4) zones of the baboon fetal adrenal obtained in late
gestation.
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) and the
fetal and definitive/transitional zones in late gestation (Fig. 6B).
The specificity for the ß isoform was evident by the relative lack of
silver grains over the adrenal cortex in sections incubated with a
sense riboprobe (Fig. 6, C and D).
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and ß proteins by immunocytochemistry protein using DAB-nickel sulfate (purple
staining, Fig. 7A) and the estrogen
receptor ß protein using DAB-imidazole (brown staining,
Fig. 8B) was observed in the baboon fetal
adrenal of late gestation. Estrogen receptor and ß
immunoreactivity was extensive in the definitive zone and somewhat less
in the transitional zone. The definitive and transitional zones were
identified by the specific staining for 3ßHSD in these two zones
(Figs. 7B and 8C). Predominant nuclear localization of the estrogen
receptor compared with cytoplasmic localization of the 3ßHSD was
observed. Consistent with mRNA expression, there was a lower level of
immunoreactivity for the estrogen receptor and ß proteins in the
fetal zone of the adrenal near term (lower halves of Figs. 7A and 8B). Specificity for estrogen receptor immunocytochemical
staining was evident by the absence of reactivity when the primary
antibody was omitted (Fig. 7C) and when the antibody was preabsorbed
with estrogen receptor protein (data not shown).
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). However, the estrogen receptor
was again expressed to a greater extent in the outer region of the
cortex. |
Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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and ß, as determined by RT-PCR, in
situ hybridization, and immunocytochemistry, were expressed in the
fetal zone and the definitive/transitional zones of the baboon fetal
adrenal gland at mid- and late gestation. Expression of estrogen
receptor and ß in the fetal zone throughout the second half of
baboon pregnancy would provide a mechanism for mediating the action of
estrogen directly within this unique cortical zone of the developing
fetal adrenal gland. We suggest, therefore, that the estrogen-dependent
suppression of ACTH-regulated DHA formation by the baboon fetal adrenal
gland (5, 6, 7) may be mediated in the classical manner by the estrogen
receptor. A direct action of estrogen on the fetal zone would
potentially provide a feedback system for modulating
C19-steroid DHA/DHAS production by the fetal adrenal gland
and thereby placental estrogen synthesis, as illustrated in Fig. 1.
Estrogen receptor and ß mRNA and protein expression appeared to
be much greater in the definitive/transitional zones than in the fetal
zone of the baboon fetal adrenal late in gestation. Moreover,
competitive RT-PCR analysis indicated that estrogen receptor mRNA
levels were much greater in the fetal adrenal of late than of
midgestation, apparently reflecting the enhanced expression of this
receptor in the definitive/transitional zones which develop in later
stages of gestation. The relatively high expression of the estrogen
receptor in the definitive/transitional zones was surprising, because
our experimental findings suggest that estrogen only indirectly
promotes maturation and function of the transitional zone (1, 3)
while simultaneously directly suppressing function of the fetal zone
(4). Further study is needed, therefore, to determine whether estrogen
may have a potentially direct action on the definitive/transitional
zones to modulate the development and/or function of the outer
compartments of the maturing fetal adrenal cortex as well.
The presence of estrogen receptor ß in the baboon fetal adrenal gland
at mid- and late gestation is consistent with the demonstration of
estrogen receptor ß mRNA by RT-PCR in the human fetal adrenal (13).
The molecular characterization, heterodimerization and interaction with
the estrogen response element (24, 25), cellular distribution, and
physiological roles of the various and ß species are only
beginning to be elucidated. Therefore, it remains to be determined
whether the action of estrogen within the primate fetal adrenal
involves interaction with either or both forms of the estrogen
receptor.
Although the specific mechanisms by which the estrogen-estrogen
receptor interaction inhibits DHA formation by the fetal zone remain to
be determined, estrogen may act to suppress the expression of the ACTH
receptor and thus cellular responsivity to ACTH. In support of this
possibility, ACTH receptor mRNA levels become decreased in the fetal
zone and elevated in the definitive/transitional zones of the baboon
fetal adrenal between mid- and late gestation in association with the
rise in estrogen in both the maternal and fetal circulations (10, 11).
Consistent with these findings in the baboon, Jaffe and co-workers (26)
have shown that ACTH receptor mRNA expression was greater in the
3ßHSD-positive definitive/transitional zones than in the fetal zone
of the human fetal adrenal and have suggested that the cells in the
outermost cortical zones are more sensitive to ACTH than are cells of
the fetal zone. Estrogen, however, may also modify ACTH
receptor-dependent secondary intracellular messengers within the fetal
adrenal, e.g. protein kinase C, which decreases the activity
of the P-450 17-hydroxylase, 17–20-lyase enzyme that catalyzes the
conversion of pregnenolone to DHA in the fetal adrenal (27). Indeed,
estrogen has been shown to regulate this enzyme in the human adrenal
(28) as well as in the rat ovary (29) and placenta (30).
In contrast to the observation of an inhibitory influence of estrogen
on DHA output as studied in short term incubates of baboon fetal
adrenal cells, in long term cultures of human fetal adrenal cells
estrogen stimulated ACTH-induced DHAS production and inhibited cortisol
synthesis by suppressing the 3ßHSD enzyme catalyzing the conversion
of DHA to 4-steroids such as cortisol (31, 32, 33). These
apparently disparate effects of estrogen may reflect a difference in
the degree of fetal adrenal cellular differentiation associated with
the different experimental conditions. For example, in tissue culture
conditions, human fetal adrenal cells develop an ultrastructure of
definitive or zona glomerulosa cells (34). Consequently, we suggest
that the apparently different responses to estrogen reported in the
literature for the human and baboon fetal adrenal do not reflect a
species difference, but, rather, appear to be due to a difference in
the qualitative nature of the cells resulting from the use of very
different in vitro experimental approaches. Regardless of
the nature of the response to estrogen, it is evident from the current
study that the estrogen receptor is expressed within the primate fetal
adrenal cortex, providing a means to mediate the action of
estrogen.
In summary, the results of the present study show that estrogen
receptor and ß were expressed in the fetal and
definitive/transitional zones of the baboon fetal adrenal cortex at
mid- and late gestation. The presence of the estrogen receptor provides
a mechanism for mediating the action of estrogen in modulating
ACTH-dependent and cortical zone-specific development and function of
the primate fetal adrenal gland.
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Acknowledgments |
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and ß. |
Footnotes |
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Received April 29, 1999.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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(ER-) and ß
(ER-ß) mRNA in the midgestational human fetus. J Clin Endocrinol
Metab 2:3509–3512
and ERß at AP1 sites. Science 277:1508–1510-hydroxylase, and
dehydroepiandrosterone sulfotransferase by adenosine 3',
5'-mono-phosphate and activators of protein kinase C in cultured human
adrenocortical cells. Endocrinology 122:2012–2018[Abstract]
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) and late (•)
gestation was plotted as a function of CRS added to each PCR reaction.
Lines were constructed by linear regression analysis, and estrogen
receptor 
200 cells) obtained by laser capture
microdissection from the fetal (lane 2) and definitive/transitional
(lanes 3 and 4) zones of the baboon fetal adrenal of late gestation.
Lane 5, Baboon endometrium; lane 1, ladder.
