| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
Original Studies |
Departments of Pathology (Y.M., T.S., H.S.), Psychosomatic Medicine (Y.M., A.T., M.H.), Internal Medicine (K.To.), and Molecular Biology (K.Ta.), Tohoku University School of Medicine, Sendai 980-8575; Department of Obstetrics and Gynecology, Yamaguchi University School of Medicine (N.S.), Ube 755-8505; and Second Division, Department of Medicine, Hamamatsu University School of Medicine (Y.O.), Hamamatsu 431-3192, Japan
Address all correspondence and requests for reprints to: Yasunari Muramatsu, M.D., Department of Pathology, Tohoku University School of Medicine, 2–1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan. E-mail: murayasu{at}patholo2.med.tohoku.ac.jp
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
were significantly higher in
the regressing corpus luteum than in the functioning corpus luteum. The
spatial and temporal immunolocalization patterns of CRF receptor were
similar to those of urocortin. These results suggest that urocortin is
locally synthesized in steroidogenic luteal cells and acts on them as
an autocrine and/or paracrine regulator of ovarian steroidogenesis,
especially during luteal regression. |
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
CRF, a 41-amino acid peptide originally isolated from the ovine hypothalamus, is generally considered to be the principal neuroregulator of the hypothalamic-pituitary-adrenal axis (12, 13). The synthesis and release of CRF has also been demonstrated in several peripheral tissues (13, 14, 15, 16, 17). The presence of immunoreactive CRF has been reported in rat and human ovaries and in human follicular fluid (18, 19). CRF and CRF-R1 mRNAs have also been detected in the thecal cells of human ovarian follicles (20). The results of several in vitro studies have demonstrated that CRF can suppress ovarian steroidogenesis (21, 22, 23). Therefore, it is suggested that the CRF system, including CRF, Ucn, and CRF receptors, may play important roles in various biological functions in the human ovary. However, Ucn and CRF-R2 expression in the human ovary have not been examined.
We previously demonstrated that the immunolocalization pattern of steroidogenic enzymes in follicles and corpora lutea were closely associated with the growth and regression of these ovarian structures (24, 25). Therefore, in the present study we examined the distribution of immunoreactive Ucn in ovarian stages classified according to the temporal and spatial expression patterns of steroidogenic enzymes in normal human ovaries throughout the menstrual cycle. Furthermore, we studied Ucn, CRF, and CRF receptor mRNA expression in corpora lutea obtained from the midluteal phase (days 19–24) and the regression phase (days 3–7) of the menstrual cycle and from early pregnant patients (6–8 weeks of pregnancy) using RT-PCR. In addition, we examined the localization of CRF and CRF receptor in the human ovary using immunohistochemistry.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Reagents
hUcn-(1–40) and r/hCRF-(1–41) were obtained from Peptide Institute (Osaka, Japan). Deoxyribonucleotide triphosphates and
Moloney murine leukemia virus reverse transcriptase were purchased from
Life Technologies, Inc. (Grand Island, NY). Random hexamer
and Taq DNA polymerase were obtained from Perkin-Elmer Corp. (Foster City, CA).
[-32P]Deoxy-CTP
([-32P]dCTP) was purchased from
Amersham Pharmacia Biotech (Arlington Heights, IL). Other
chemicals used in this study were obtained from Katayama Chemical, Inc. (Osaka, Japan) and Wako Pure Chemical Industries, Ltd. (Osaka, Japan).
Tissue collections for immunohistochemistry
Normal cycling human ovaries (n = 24) were obtained from patients who underwent oophorectomy and hysterectomy. All patients were diagnosed with squamous cell carcinoma of the uterine cervix. Oophorectomy was performed to rule out possible foci of metastasis in the ovary. Histopathological examination of these ovaries was performed in hematoxylin-eosin-stained tissue sections. No morphological abnormalities were observed in these sections, including metastasis of the carcinoma or polycystic changes. The age of the patients ranged from 26–45 yr, and the patients had regular menstrual cycles and no sex steroid abnormalities before surgery. Ovarian tissues were immediately fixed in 4% paraformaldehyde (pH 7.4) for 12 h at 4 C and were subsequently embedded in paraffin wax.
The phase of the menstrual cycle was determined in each case by taking a detailed patient history and performing endometrial dating based on the criteria of Silverberg (26). In addition, serum levels of estradiol and progesterone were determined using an enzyme-linked immunosorbent assay kit (Diagnostic Products, Los Angeles, CA) at the time of surgery. Ovarian follicles and corpora lutea for each specimen were classified according to the phase of the menstrual cycle, morphological features, and immunolocalization patterns of steroidogenic enzymes, as reported by Suzuki et al. (24, 25).
Seven follicular stages were classified in the present study as
follows: primordial follicles (n = 54), primary follicles (n
= 40), preantral follicles (n = 16), nondominant antral follicles
in the follicular phase [P450 aromatase (P450arom)-negative, P450
cholesterol side-chain cleavage (P450scc)-positive, 3ß-hydroxysteroid
dehydrogenase (3ßHSD)-positive, and P450 17-hydroxylase
(P450c17)-positive antral follicles in the follicular phase; n =
13], dominant follicles (all enzyme-positive antral follicles in the
follicular phase; n = 7), nondominant antral follicles in the
luteal phase (P450arom-negative, P450scc-positive, 3ßHSD-positive,
and P450c17-positive antral follicles in the luteal phase; n =
12), atretic follicles (n = 24), and oocytes (n = 57).
Six luteal stages were identified as follows: early corpus luteum (P450scc-positive, 3ßHSD-positive, P450arom-positive, and P450c17-positive functioning corpus luteum in the early luteal phase; n = 4), midcorpus luteum (all enzyme-positive functioning corpus luteum in the mid luteal phase; n = 6), late corpus luteum (all enzyme-positive functioning corpus luteum in the late luteal phase; n = 6), early degenerating corpus luteum (P450arom-negative, P450scc-positive, 3ßHSD-positive and P450c17-positive corpus luteum; n = 8), late degenerating corpus luteum (all enzyme-negative corpus luteum; n = 12), and corpus albicans (n = 34).
Corpora lutea of early pregnancy (6–8 weeks of pregnancy; n = 3) were also obtained from patients, aged 24–30 yr, with ectopic pregnancy. These specimens were immediately fixed in 4% paraformaldehyde (pH 7.4) for 12 h at 4 C and subsequently embedded in paraffin wax. The pregnant corpus luteum showed the same immunolocalization patterns of steroidogenic enzymes as the functioning corpus luteum (i.e. all enzymes were positive).
Antibodies
r/hUcn-(21–35) (ARTQSQRERAEQNRI) was obtained from Iwaki Glass Co. (Funabashi, Japan), and rUcn1–40 was purchased from Sawady Technology (Tokyo, Japan). A specific antiserum against Ucn was raised in a rabbit immunized with a peptide corresponding to r/hUcn-(21–35). Methods of immunization and characterization of the antiserum were previously reported (27). An antiserum against rUcn-(1–40) (no. 1381) was also raised in a rabbit (9). Neither of the two Ucn antisera cross-reacted with other peptides, including human CRF and urotensin I (9, 27).
A rabbit polyclonal antibody against r/hCRF was provided by Dr. Mouri (28). Cross-reacion of this antibody with human Ucn and urotensin I was less than 0.001% (9, 28).
An antiserum against CRF receptor (CRF-R1 C-20) was obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA). The CRF receptor antibody was raised in a goat against a peptide corresponding to amino acids 425–444 mapping at the carboxyl-terminus of hCRF-R1. This antibody reacts with both CRF-R1 and CRF-R2 (29).
To characterize ovarian function in detail, we used a polyclonal antibody for Ad4-binding protein (Ad4BP; provided by Dr. K. Morohashi, Kyushu University, Fukuoka, Japan). Ad4BP has been identified as a steroidogenic cell-specific transcription factor that activates transcription of steroidogenic P450 genes and has been suggested to have the potential to regulate the expression of steroidogenic enzymes in the human ovary (30).
Immunohistochemistry
Immunohistochemical analysis was performed employing the streptavidin-biotin amplification method using a Histofine Kit (Nichirei, Tokyo, Japan). After deparaffinization, slides were heated in an autoclave at 120 C for 5 min in citric acid buffer (2 mmol/L citric acid and 9 mmol/L trisodium citrate dehydrate, pH 6.0) for anti-r/hUcn-(21–35), anti-rUcn-(1–40), anti-r/hCRF, and Ad4BP immunostaining. The dilutions of the primary antibodies used in our study were as follows: anti-r/hUcn-(21–35), 1:2500; anti-rUcn-(1–40), 1:1000; anti-r/hCRF, 1:1000; anti-CRF-R, 1:20; and Ad4BP, 1:700. The antigen-antibody complex was visualized with 3,3'-diaminobenzidine solution [1 mmol/L 3,3'-diaminobenzidine, 50 mmol/L Tris-HCl buffer (pH 7.6), and 0.006% H2O2] and counterstained with hematoxylin. As a negative control, preabsorbed antiserum and/or 10 mmol/L PBS was used instead of primary antibodies.
For all absorption tests of immunoreactivity, an antibody-antigen mixture containing an optimally diluted antiserum, which is equal to a peptide solution, including hUcn-(1–40) or r/hCRF-(1–41) in a volume of 20 µmol/L (final peptide concentration), was incubated at 4 C for 1 night. After centrifugation, the resultant supernatants were used as preabsorbed antibodies (8, 10).
Scoring of immunoreactivity
Immunohistochemical procedures were performed at least twice for each tissue section for confirmation of immunolocalization. Three of the authors (Y.M., T.S., and H.S.) independently graded the ovarian follicles and corpora lutea for each specimen and separated them into the following three groups: 2+, strongly positive; +, weakly positive; and -, negative. Disconcordant results among the observers were simultaneously reevaluated using multiheaded light microscopy.
Corpora lutea used for RT-PCR analysis
Corpora lutea used for RT-PCR analysis were obtained at hysterectomy from normal cycling women, aged 39–49 yr, who underwent surgery for myoma uteri or cervical cancer. Menstrual history and endometrial dating, diagnosed histologically according to the criteria of Silverberg (26), were used to determine the phase of the corpus luteum. Corpora lutea were obtained from the midluteal phase (days 19–24; n = 3) and the regression phase (days 3–7; n = 3) with day 1 being marked by the onset of menstruation. Corpora lutea of early pregnancy (6–8 weeks of pregnancy; n = 3) were obtained from patients, aged 24–30 yr, with ectopic pregnancy. Tissue samples were washed with saline to remove blood, immediately frozen in liquid nitrogen, and stored at -80 C until RNA isolation.
RT-PCR
Total RNA was isolated from corpora lutea with Isogen
(Wako Pure Chemical Industries, Ltd.) using the method
provided by the manufacturer. For mRNA analysis, RT-PCR was performed
as previously reported (31). Sequences of the primers are
summarized in Table 1
. Direct sequence
analyses of the PCR products were performed for sequence verification.
Each reaction also included primers (5'-CTGAAGGTCAAAGGGAATGTG-3' and
5'-GGACAGAGTCTTGATGATCTC-3') to amplify ribosomal protein L19 or
primers (5'-CGTTCACCTTGATGAGCCCATT-3' and 5'-TCCAAGGGTCCGCTGCAGTC-3')
to amplify ribosomal protein S16. Both L19 and S16 were used as
internal controls (36, 37). The predicted size of the
PCR-amplified product was 194 bp for L19 and 100 bp for S16. In brief,
3 µg total RNA were reverse transcribed at 42 C in a reaction mixture
(single strength PCR buffer, 2.5 mmol/L deoxynucleotide triphosphates,
5 µmol/L random hexamer primer, 1.5 mmol/L
MgCl2, and 200 U Moloney murine leukemia virus
reverse transcriptase). The RT product was aliquoted equally into two
tubes for Ucn, CRF, CRF-R1, CRF-R2 or CRF-R2ß primers and internal
control primers (L19 or S16). For PCR amplification, a mixture
containing the oligonucleotide primers (50 pmol),
[-32P]deoxy-CTP (2 µCi at 3000 Ci/mmol),
and Taq DNA polymerase (2.5 U) was added to each reaction.
Amplification was carried out for 32 cycles at 94 C (1 min), 62 C (1
min), and 72 C (1 min) for Ucn; 32 cycles at 94 C (1 min), 70 C (1
min), and 72 C (1 min) for CRF; and 35 cycles at 94 C (1 min), 65 C (1
min), and 72 C (1 min) for CRF-R1, CRF-R2, and CRF-R2ß, followed
by 10 min of final extension at 72 C in a programmed temperature
control system (PC-800, ASTEC, Fukuoka, Japan). Reaction products were
electrophoresed on an 8% polyacrylamide nondenaturing gel. After
autoradiography, band intensities were analyzed using a bioimaging
analyzer BAS 2000 (Fuji Photo Film Co., Ltd., Tokyo,
Japan). For quantification, the intensity of Ucn, CRF, CRF-R1,
CRF-R2, or CRF-R2ß was normalized to that of the internal control
L19 or S16. All samples were subjected to RT-PCR at the same time.
|
Data were examined by ANOVA and Duncan’s new multiple range test. Differences were considered significant at P < 0.05.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Immunoreactivity for Ucn was present in the cytoplasm, and that of Ad4BP was detected in the nuclei. There were no differences in the immunolocalization patterns between anti-r/hUcn-(21–35) and anti-rUcn-(1–40) antibodies. The distribution of Ad4BP in the ovaries was in good agreement with the report by Takayama et al. (30).
In dominant follicles, weak immunoreactive Ucn was detected in both
granulosa and theca interna cells (Fig. 1A). Weak immunoreactivity was also
detected in theca interna cells of nondominant follicles (in both the
follicular and luteal phases) and in atretic follicles, but not in
granulosa cells (Fig. 1C). No significant immunoreactivity for Ucn was
detected in primordial, primary, or preantral follicles or oocytes.
|
. After ovulation, immunoreactive Ucn
was detected in luteinized granulosa and thecal cells in the midcorpus
luteum (Fig. 2A) and late corpus luteum.
In the early degenerating corpus luteum, intense Ucn immunoreactivity
was detected in luteinized thecal cells, whereas no immunoreactivity
for Ucn was detected in luteinized granulosa cells (Fig. 3A). Ad4BP was present only in luteinized
thecal cells (Fig. 3C). Ucn immunoreactivity in luteinized thecal cells
was relatively weak in the late degenerating corpus luteum and was
negative in the corpus albicans. In the pregnant corpus luteum, weak
Ucn immunoreactivity was detected in luteinized granulosa and thecal
cells. All Ucn immunoreactivity described above was prevented by
preabsorption with corresponding synthetic peptides hUcn-(1–40) (Figs. 1B
, 2B, and 3B), but not with r/hCRF-(1–41).
|
|
|
In dominant follicles, immunoreactive CRF was detected in theca interna cells. Weak CRF-R immunoreactivity was detected in both granulosa and theca interna cells of dominant follicles. No significant CRF or CRF-R immunoreactivity was detected in primordial, primary, preantral, nondominant, or atretic follicles or oocytes.
The results of our CRF immunohistochemical study in the corpus luteum
are summarized in Table 2. Immunolocalization patterns for CRF in the
corpus luteum were almost the same as those for Ucn described above.
Weak CRF immunoreactivity was observed in luteinized granulosa and
thecal cells of the functioning corpus luteum (Fig. 2C) and pregnant
corpus luteum. In the early degenerating corpus luteum, intense
immunoreactivity for CRF was detected predominantly in luteinized
thecal cells, and weak CRF immunoreactivity was also detected in
luteinized granulosa cells (Fig. 3D).
The results of the immunohistochemical study for CRF-R in the corpus
luteum are summarized in Table 2. Immunolocalization patterns for CRF-R
were similar to those of CRF-R agonists (i.e. Ucn and CRF).
In the early degenerating corpus luteum, where both Ucn and CRF
immunoreactivities were predominantly detected in luteinized thecal
cells, CRF-R immunoreactivity was detected in luteinized thecal cells
(Fig. 3E).
RT-PCR analysis of corpora lutea
RT-PCR analyses were performed three times. Representative data
are shown in
Figs. 4–7.
RT-PCR analyses revealed that mRNA levels for Ucn, CRF, CRF-R1, and
CRF-R2 were significantly higher in the regressing corpus luteum
than in the midluteal phase and pregnant corpus luteum, and that mRNA
for CRF-R2ß was not detected. There was no significant difference
between the expression of these genes in the midluteal phase of the
menstrual cycle and early pregnancy.
|
|
|
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Mastrakos et al. (19) also reported the presence of immunoreactive CRF in the human corpus luteum. In their study CRF immunoreactivity was detected in small theca-derived and large granulosa-derived luteinized cells of developing corpora lutea, but immunoreactive CRF was less prominent or was totally absent in regressing corpora lutea. Our RT-PCR analyses demonstrated that mRNA levels for CRF were significantly higher in the regressing corpus luteum than in the midluteal phase or pregnant corpus luteum. We also demonstrated that immunoreactive CRF was markedly present in luteinized thecal cells of the early degenerating corpus luteum and that weak CRF immunoreactivity was also detected in luteinized granulosa cells of the early degenerating corpus luteum. In the functioning corpus luteum (mid and late luteal phases) and pregnant corpus luteum, weak immunostaining for CRF was detected in luteinized granulosa and thecal cells. The results from our immunohistochemical studies were consistent with our RT-PCR findings. The differences between the present study and the report by Mastrakos et al. may be due to the differences in the antibodies employed or other factors. Further investigations are required to clarify these discrepancies.
Ucn has been demonstrated to bind with both CRF-R1 and CRF-R2. In
CRF-R1-transfected cells, Ucn and CRF both function to induce cAMP
levels in the same concentration range, with dissociation constants
(Ki) of 0.41 and 0.95 nmol/L, respectively. Ucn
is at least 10-fold more potent than CRF in stimulating cAMP
accumulation in CRF-R2-transfected cells (6). RT-PCR
analyses in our study demonstrated that CRF-R1 and CRF-R2 mRNA
levels were significantly higher in the regressing corpus luteum than
in the midluteal phase and pregnant corpus luteum. CRF-R2ß mRNA was
undetectable in all corpora lutea samples examined by RT-PCR analyses
in the present study. The absence of CRF-R2ß is consistent with the
fact that this subtype of CRF-R2 is generally considered to be a minor
isoform in human tissues (34, 35). In the regressing
corpus luteum, which was shown to express prominent levels of CRF-R1
and CRF-R2, CRF-R was immunolocalized only to luteinized thecal
cells, whereas no CRF-R immunoreactivity was detected in luteinized
granulosa cells. In the functioning and pregnant corpus luteum, CRF-R
immunoreactivity was detected in luteinized granulosa and thecal cells.
The spatial and temporal colocalization of CRF receptors and their
agonists (i.e. Ucn and CRF) suggests that Ucn and CRF may
act as autocrine and/or paracrine regulators in the ovarian corpus
luteum, most likely by inhibiting ovarian steroidogenesis in the
process of luteal degeneration.
CRF and CRF-R1 expression in human ovarian follicles have been investigated by Mastrakos et al. (19) and Asakura et al. (20). CRF protein and mRNA have been shown to be localized in the thecal cells of follicles (19, 20). CRF-R1 mRNA signals have been reported in thecal cells of follicles. Granulosa cells were devoid of CRF and CRF-R1 mRNA and protein (20). In our immunohistochemical study, Ucn, CRF, and CRF-R were weakly detected in thecal cells of dominant follicles, but the presence of these immunoreactivities in thecal cells of ovarian follicles was consistent with previous investigations. Jacobs et al. (38) and Mastrakos et al. (19) reported that no immunoreactive CRF was present in oocytes of the human ovary. We also did not detect Ucn, CRF, or CRF-R immunoreactivity in oocytes, which was consistent with the findings of previous reports.
In the functioning corpus luteum, which is actively involved in the production and secretion of sex steroids, both P450scc and 3ßHSD are highly expressed in luteinized thecal and granulosa cells. Almost all cells also express Ad4BP. P450arom is expressed in luteinized granulosa cells, and P450c17 is present in luteinized thecal cells. After menstruation, the functioning corpus luteum starts to degenerate. Degenerating corpora lutea can be classified into early (steroid-producing) degenerating corpora lutea and late (nonsteroid-producing) degenerating corpora lutea. Corpora lutea in the early degenerating phase have been demonstrated to express P450scc, 3ßHSD, P450c17, and Ad4BP only in luteinized thecal cells. Luteinized granulosa cells are senescent and thus lose their steroidogenic function or Ad4BP expression. Subsequently, luteinized thecal cells in the late degenerating corpus luteum become quiescent in both the process of steroidogenesis and cell proliferation (24, 30, 39, 40). Macrophages and/or T lymphocytes have been shown to participate in luteal regression. The numbers of macrophages and T lymphocytes are highest in the late degenerating corpus luteum (41, 42). The peak of Ucn, CRF, and CRF receptor expression occurred during the stage of regression, earlier than the appearance of leukocytes. The spatial and temporal localization of Ucn, CRF, and CRF receptors correlated well with that of Ad4BP. In the functioning and pregnant corpus luteum, Ucn, CRF, CRF-R, and Ad4BP were all detected in luteinized granulosa and thecal cells. In the early degenerating corpus luteum, they were detected only in luteinized thecal cells, whereas no significant immunoreactivity was present in luteinized granulosa cells. Ad4BP is a steroidogenic cell-specific transcription factor that activates the transcription of steroidogenic P450 genes, including P450scc, P450c17, and P450arom (30). Therefore, our results suggest that Ucn and CRF are produced in steroidogenic luteal cells and act on them in a paracrine/autocrine/intracrine fashion. Locally synthesized Ucn and CRF may be involved in ovarian steroidogenesis. Furthermore, marked expression of Ucn, CRF, and CRF receptors in the regressing corpus luteum suggests that Ucn and CRF play important roles in luteal regression rather than in the maintenance of the functioning or pregnant corpus luteum.
Several studies have reported that CRF can suppress ovarian steroidogenesis in vitro (21, 22, 23). Both Calogero et al. (21) and Ghizzoni et al. (22) have isolated human granulosa-luteal cells from the follicular fluid upon oocyte retrieval. These patients were undergoing in vitro fertilization and had received a GnRH analog and FSH therapy before oocyte retrieval. These two groups of investigators demonstrated that CRF exerts an inhibitory effect on estrogen and progesterone production in cultured human granulosa-luteal cells. This effect was mediated via CRF and interleukin-1 receptor, and was independent of adenylate cyclase-cAMP generation (21, 22). Erden et al. (23) isolated follicular thecal cells from patients undergoing benign gynecological surgery during the follicular phase and demonstrated that CRF inhibited LH-stimulated dehydroepiandrosterone and androstenedione production in isolated thecal cells. In addition, they found that CRF reduced LH-stimulated P450c17 mRNA levels. CRF and CRF receptors in the present investigation were demonstrated to be predominantly expressed in luteinized thecal cells of the early degenerating corpus luteum, which were losing their steroidogenic function. This finding may be one of the pieces of evidence that suggests that CRF has the capacity to suppress ovarian steroidogenesis in vivo as it does in vitro. Moreover, Ucn, like CRF, may also suppress ovarian steroidogenesis. However, to date there have been no in vitro studies looking at Ucn regulation of ovarian steroidogenesis. Furthermore, studies concerning CRF regulation have been limited to the use of follicular cells or cell components within the follicular fluid. There have also been no studies on CRF or Ucn using corpus luteum components. Further studies are necessary to clarify the biological roles of Ucn and CRF in the human ovarian corpus luteum.
|
Acknowledgments |
|---|
|
Footnotes |
|---|
Received June 27, 2000.
Revised October 16, 2000.
Accepted November 8, 2000.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
This article has been cited by other articles:
 |
|
 |
|
  A. Yata, K. Nakabayashi, S. Wakahashi, N. Maruo, N. Ohara, and T. Maruo Suppression of progesterone production by stresscopin/urocortin 3 in cultured human granulosa-lutein cells Hum. Reprod., July 1, 2009; 24(7): 1748 - 1753. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Xu, F. Xu, J. D. Hennebold, T. A. Molskness, and R. L. Stouffer Expression and Role of the Corticotropin-Releasing Hormone/Urocortin-Receptor-Binding Protein System in the Primate Corpus Luteum during the Menstrual Cycle Endocrinology, November 1, 2007; 148(11): 5385 - 5395. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Mastorakos, E. I Karoutsou, and M. Mizamtsidi Corticotropin releasing hormone and the immune/inflammatory response Eur. J. Endocrinol., November 1, 2006; 155(suppl_1): S77 - S84. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Xu, J. D. Hennebold, and R. L. Stouffer Dynamic Expression and Regulation of the Corticotropin-Releasing Hormone/Urocortin-Receptor-Binding Protein System in the Primate Ovary during the Menstrual Cycle J. Clin. Endocrinol. Metab., April 1, 2006; 91(4): 1544 - 1553. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. R. Lindsay and L. K. Nieman The Hypothalamic-Pituitary-Adrenal Axis in Pregnancy: Challenges in Disease Detection and Treatment Endocr. Rev., October 1, 2005; 26(6): 775 - 799. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Fukuda, K. Takahashi, T. Suzuki, M. Saruta, M. Watanabe, T. Nakata, and H. Sasano Urocortin 1, Urocortin 3/Stresscopin, and Corticotropin-Releasing Factor Receptors in Human Adrenal and Its Disorders J. Clin. Endocrinol. Metab., August 1, 2005; 90(8): 4671 - 4678. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Xu, R.L. Stouffer, R.P. Searles, and J.D. Hennebold Discovery of LH-regulated genes in the primate corpus luteum Mol. Hum. Reprod., March 1, 2005; 11(3): 151 - 159. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. C. Reubi, B. Waser, W. Vale, and J. Rivier Expression of CRF1 and CRF2 Receptors in Human Cancers J. Clin. Endocrinol. Metab., July 1, 2003; 88(7): 3312 - 3320. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. D. Catalano, T. Kyriakou, J. Chen, A. Easton, and E. W. Hillhouse Regulation of Corticotropin-Releasing Hormone Type 2 Receptors by Multiple Promoters and Alternative Splicing: Identification of Multiple Splice Variants Mol. Endocrinol., March 1, 2003; 17(3): 395 - 410. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Kimura, K. Takahashi, K. Totsune, Y. Muramatsu, C. Kaneko, A. D. Darnel, T. Suzuki, M. Ebina, T. Nukiwa, and H. Sasano Expression of Urocortin and Corticotropin-Releasing Factor Receptor Subtypes in the Human Heart J. Clin. Endocrinol. Metab., January 1, 2002; 87(1): 340 - 346. [Abstract] [Full Text] [PDF]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| Endocrinology | Endocrine Reviews | J. Clin. End. & Metab. |
| Molecular Endocrinology | Recent Prog. Horm. Res. | All Endocrine Journals |
