Production of activin A in hyperplasia and adenocarcinoma of the human endometrium.

OBJECTIVES: To examine the possible localization and production of activin A in human normal endometrium, endometrial hyperplasia, and adenocarcinoma tissues. METHODS: Human endometrial tissues were collected from 45 patients who were undergoing abdominal hysterectomy. Tissue sections were stained with monoclonal antibodies against the inhibin/activin alpha- and beta A-subunits and activin A using an avidin-biotin-peroxidase complex technique. Concentrations of activin A and inhibin A in tissue extracts of the endometrial tissues were measured using enzyme-linked immunosorbent assays (ELISAs). The expressions of the inhibin alpha-subunit and activin beta A-subunit messenger RNA (mRNA) in the endometrial tissues were demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) analysis. RESULTS: No immunostaining with an antibody against the inhibin alpha-subunit was observed in human normal endometrium, endometrial hyperplasia, and adenocarcinoma. By contrast, immunostaining for the activin beta A-subunit and activin A was observed in the cytoplasm of glandular cells in normal endometrium, endometrial hyperplasia, and tumor cells of endometrial adenocarcinoma. The percentages of stained cells in endometrial adenocarcinoma were higher than those in normal endometrium. Also, the percentages of stained tumor cells with poor differentiation were higher than those with good and moderate differentiation of the endometrium. The stromal cells in normal endometrium, endometrial hyperplasia, and adenocarcinoma were weakly immunoreactive with antibodies against the beta A-subunit and activin A. Immunoreactivity of activin A in tissue extracts from normal endometrium and endometrial adenocarcinoma was detected by the two-site ELISA. Immunoreactivity of activin A was significantly higher in adenocarcinoma than in normal endometrium. On the other hand, the immunoreactive inhibin A was not detected. The expression of the alpha-subunit mRNA in endometrial tissues was demonstrated as the RT-PCR products migrated at 905 bp and the PCR products of the beta A-subunit showed a band at 366 bp. CONCLUSIONS: It is suggested that activin A, but not inhibins, are produced by endometrial tissues. The amounts of produced activin A were higher in adenocarcinoma tissues than in normal endometrium. Activin A might be involved in human endometrial tumorigenesis.

Apoptosis and the expression of Bax and Bcl-2 in hyperplasia and adenocarcinoma of the uterine endometrium.

BACKGROUND: Apoptosis plays a crucial role in carcinogenesis in various tumours. This study was designed to investigate the occurrence of apoptosis and the expression of Bcl-2 and Bax proteins in endometrial tumours of corpus uteri. METHODS: Endometrial tissues were obtained from 20 patients with endometrioid adenocarcinoma, 16 patients with endometrial hyperplasia, and 4 patients with myoma uteri (which were used as controls). The occurrence of apoptosis was examined by using molecular biochemical techniques. The expression of Bcl-2 and Bax proteins was also investigated using immunohistochemical staining with appropriate antibodies. RESULTS: The labelling of DNA in situ indicated that apoptotic cells were sporadically seen in postmenopausal endometrium (5.2 +/- 2.1, n = 4) and endometrial hyperplasia without atypia (2.6 +/- 0.5, n = 9). In contrast, labelled cells were detected in atypical endometrial hyperplasia (15.9 +/- 2.2, n = 7), and their numbers increased intensely in adenocarcinoma (29.3 +/- 3.7, n = 20). Autoradiographic analysis revealed DNA laddering in many cases of carcinoma. Bcl-2 was highly immunopositive in hyperplasia without atypia (36.2 +/- 6.5%, n = 9), but was decreased in the atypical endometrial hyperplasia (16.3 +/- 4.8%, n = 7). Large fractions of the carcinoma (6.3 +/- 1.8%, n = 20) and normal endometrium (2.8 +/- 1.4%, n = 4) were immunonegative or slightly immunopositive to Bcl-2. In contrast, Bax immunoreactivity was more frequent and stronger in adenocarcinoma (43.6 +/- 4.1%, n = 20) than that in normal endometrium (17.6 +/- 6.7%, n = 4) and hyperplasia (7.2 +/- 2.2%, n = 16). CONCLUSIONS: These results suggest that cells in hyperplasia expressing Bcl-2 might have prolonged survival ability. Neoplastic cells in adenocarcinoma might show apoptosis in association with a decreased expression of Bcl-2 and an increased expression of Bax. Therefore, the frequency of apoptosis and the expression of Bcl-2 and Bax might be correlated with carcinogenesis in the uterine endometrium of humans.

Apoptosis and the expression of Bcl-2 and Bax in patients with endometrioid, clear cell, and serous carcinomas of the uterine endometrium.

OBJECTIVES: Clear cell and serous carcinoma of the uterus are rare types of endometrial carcinomas. This study was designed to investigate the differential occurrence of apoptosis, Bcl-2, and Bax in endometrioid, clear cell, and serous carcinomas. METHODS: In a total of 28 endometrial carcinomas as well as 4 samples of normal postmenopausal endometria, apoptotic changes were examined using molecular biochemical techniques. The expression of Bcl-2 and Bax proteins was also investigated by immunohistochemical staining with appropriate antibodies. RESULTS: Labeling of DNA in situ indicated that apoptotic cells were sporadically seen in postmenopausal endometrium (5.2 +/- 2.1, n = 4). In contrast, cells undergoing apoptosis apparently were detected in endometrioid carcinoma (29.3 +/- 3.7, n = 20), and their numbers increased intensely in clear cell (49.5 +/- 5.6, n = 5) and serous carcinomas (50.8 +/- 6.0, n = 3). Autoradiographic analysis revealed that high-molecular-weight DNA was predominant in postmenopausal endometrium. However, a DNA ladder was identified in 7 of 10 carcinomas. Although Bcl-2 was immunonegative or faintly immunopositive in all cases, many cases of endometrioid carcinoma (43.6 +/- 4.1%, n = 20) were immunopositive for Bax, unlike postmenopausal endometrium (17.6 +/- 6.7%, n = 4). Moreover, the number of cells expressing Bax increased in clear cell (60.4 +/- 6.5%, n = 5) and serous carcinomas (66.8 +/- 7.6%, n = 3) compared with that in endometrioid carcinoma. CONCLUSIONS: These results indicate that apoptosis occurs in a specific population of cells in different histologic components of endometrial carcinomas. The expression of Bax, but not of Bcl-2, might suggest histologic differentiation in endometrial carcinomas.

The vascular endothelial growth factor/fms-like tyrosine kinase system in human ovary during the menstrual cycle and early pregnancy.

In human ovaries, angiogenesis is known to be associated with the development of follicles and the formation of the corpus luteum (CL). A complex vascular network is formed within the thecal cell layer during follicular growth, and rapid neovascularization occurs toward the granulosa cell layer after ovulation. Vascular endothelial growth factor (VEGF) is a multifunctional cytokine, stimulating endothelial cell growth and enhancing microvascular permeability. A specific receptor for VEGF, fms-like tyrosine kinase (Flt-1), is expressed in vascular endothelial cells that mediates the action of VEGF. We examined the localization and expression of VEGF and Flt-1, using an immunohistochemical technique and RT-PCR analysis, in human follicles and corpora lutea during the normal menstrual cycle and early pregnancy. We measured concentrations of VEGF in extracts of human CL using an enzyme-linked immunosorbent assay during the luteal phase and early pregnancy. Immunostaining for VEGF was observed in granulosa cells from small antral follicles to preovulatory follicles. The staining was detected in thecal cells from medium-sized to preovulatory follicles. The intensity of the staining was gradually increased as a follicle grew. Flt-1 was localized in granulosa and thecal cells of preovulatory follicles as well as in endothelial cells. In the human CL, the intense staining for VEGF was observed in granulosa and thecal lutein cells, especially in the midluteal phase. The immunostaining for Flt-1 was faint in endothelial cells in the CL, whereas it was distinct in granulosa and thecal lutein cells. The concentrations of VEGF in lutein extracts were high in the early and midluteal phases and tended to decrease toward the late luteal phase. During early pregnancy, a measurable amount of VEGF was detected. RT-PCR analysis demonstrated that messenger ribonucleic acids encoding VEGF121, VEGF165, and Flt-1 were expressed in the CL. These results suggest that VEGF might have an autocrine role in the ovulatory process and luteal function as well as a paracrine role in angiogenesis.

Immunohistochemical localization of inhibin and activin subunits in human epithelial ovarian tumors.

OBJECTIVE: Our purpose was to examine the cellular localization of inhibin and activin subunits in human epithelial ovarian tumors. STUDY DESIGN: We examined the immunohistochemical localization of the alpha, betaA, and betaB subunits of inhibin in human mucinous and serous ovarian tumors including adenoma, cystic tumor with borderline malignancy, and adenocarcinoma. RESULTS: Immunostaining specific for the alpha, betaA, and betaB subunits of inhibin was observed in the tumor cells of the mucinous adenoma and the cystic tumor with borderline malignancy. We observed negative immunostaining specific for the alpha subunit and positive staining specific for the betaA and betaB subunits in the tumor cells of the mucinous adenocarcinoma. We did not observe any staining for the alpha subunit of inhibin in the serous tumors including benign adenoma, cystic tumor with borderline malignancy, and adenocarcinoma. However, positive staining results for the betaA and betaB subunits were observed in the serous tumor cells. CONCLUSION: Our results suggest that inhibins and activins might be secreted by the mucinous adenoma and the cystic tumor with borderline malignancy and that activins might be secreted by the mucinous adenocarcinoma and the serous tumors including benign adenoma, cystic tumor with borderline malignancy, and adenocarcinoma.

The role of tyrosine kinase in gonadotropin-induced ovulation in the rat ovary.

OBJECTIVES: Our purpose was to elucidate the involvement of the tyrosine kinase pathway in gonadotropin-induced ovulation in the rat ovary. STUDY DESIGN: We investigated the effect of a tyrosine kinase inhibitor, tyrphostin, on the rat ovulatory process in vivo and in vitro. METHODS: In cultured rat granulosa cells, the effect of tyrphostin on LH-, dibutyryl cyclic AMP ((Bu)2cAMP)- or forskolin-stimulated tissue type plasminogen activator (tPA) activities was examined by using a fibrin autography technique. In an in vivo system, tyrphostin was injected into the bursal cavity of the ovary in pregnant mare serum gonadotropin-treated rats, just before human chorionic gonadotropin administration. After 24 h, the number of oocytes in the oviduct was counted and the tyrphostin-treated ovaries were examined histologically. RESULTS: Tyrphostin inhibited LH-stimulated tPA activity but did not affect (Bu)2cAMP- or forskolin-stimulated ones. In an in vivo study, tyrphostin suppressed oocyte release dose-dependently. Histological observations revealed that tyrphostin-treated ovaries contained many large unruptured follicles and a few corpora lutea. CONCLUSION: This study suggests that the suppressive effect of tyrphostin on ovulation may be partly due to tPA activity inhibition in the granulosa cells via the suppression of tyrosine kinase activity. Additionally, tyrosine kinase phosphorylation may be involved in gonadotropin-activated signaling systems in the rat ovulatory process.

Immunohistochemical localization of activin A in human endometrial tissues during the menstrual cycle and in early pregnancy.

OBJECTIVE: To investigate the possible localization of activin A in human endometrial tissue. METHODS: Human endometrial tissue was collected from 33 patients who were undergoing abdominal hysterectomy. Human decidual tissue was collected from 11 patients, who were having a therapeutic abortion. Tissue was fixed in Bouin's solution and made into paraffin sections. Tissue sections were stained with monoclonal antibodies against the inhibin/activin alpha- and betaA-subunits and activin A using an avidin-biotin-peroxidase complex technique. RESULTS: No immunostaining with antibody against the alpha-subunit was observed in the human endometrium during the menstrual cycle or in the decidua during early pregnancy. By contrast, immunostaining for the betaA-subunit and activin A was observed in the cytoplasm of endometrial glands at all phases of the menstrual cycle and in the decidua during early pregnancy. The intensity of immunostaining for the betaA-subunit was strong during the menstrual phase, became weaker during the early proliferative phase, and was intense again at the late proliferative phase. The immunostaining for the betaA-subunit was weak during the early secretory phase and became very intense toward the midsecretory and late secretory phases. The intensity of immunostaining for activin A changed during the menstrual cycle and showed a tendency similar to that for betaA-subunit. The stromal cells were weakly immunoreactive with antibodies against the betaA-subunit and activin A from the menstrual to the midsecretory phase and became strong in the late secretory phase. Intense staining for the betaA-subunit and activin A was observed in the cytoplasm of decidual cells during early pregnancy. CONCLUSION: Activin A, but not inhibins, is localized in the endometrial tissue. The endometrium may be a major source of activin A during the normal menstrual cycle, and the decidua may be one of the sources of activin A during early pregnancy.

[Anaphylactoid shock caused by chlorhexidine gluconate].

We report two cases of anaphylactoid shock caused by chlorhexidine gluconate. Both patients had skin flare, severe hypotension and increased airway pressure during cannulation of an antibacterial IVH catheter containing chlorhexidine gluconate after skin sterilization with chlorhexidine gluconate. In case 1, we did not identify the mechanism and causative drugs. In case 2, an intradermal test to chlorhexidine gluconate was positive 2 months later. Then we confirmed that the anaphylactoid shock was caused by chlorhexidine gluconate. We should bear in mind the risk of anaphylactoid shock when we use chlorhexidine gluconate or the IVH catheter containing the bactericide.

Serum levels of inhibin A and inhibin B in women with normal and abnormal luteal function.

OBJECTIVE: To determine whether serum inhibin A and inhibin B concentrations are lower in patients with luteal dysfunction than in women with normal luteal function. METHODS: Serum samples were collected from seven healthy women with regular menstrual cycles. Serum samples on days +5 to +9 after the LH surge were collected from patients with luteal dysfunction. The diagnosis of luteal dysfunction was based on a luteal phase duration less than 11 days and a single midluteal progesterone level below 10 ng/mL. Serum levels of inhibin A, inhibin B, progesterone, estradiol (E2), FSH, and LH were measured. RESULTS: The serum inhibin A levels were increased toward the late follicular phase. The levels reached a maximum during the midluteal phase, followed by a fall during the late luteal phase. The serum inhibin B levels were high during the follicular phases and the early luteal phase. The levels decreased during the midluteal and late luteal phases. Serum levels (mean +/- standard error of the mean) of inhibin A in patients with luteal dysfunction were significantly lower than those in women during the midluteal phase (26.2 +/- 2.9 compared to 41.9 +/- 2.8 pg/mL; P < .01) in addition to the expected decrease in serum progesterone levels (6.3 +/- 0.7 compared to 14.7 +/- 1.2 ng/mL; P < .01). Serum inhibin B levels did not differ significantly between normal women and those with luteal dysfunction. There also were no significant differences in the E2, FSH, and LH levels. CONCLUSION: Levels of inhibin A, but not of inhibin B, may reflect the human luteal function.

Apoptosis of human ovary and uterine endometrium during the menstrual cycle.

To investigate apoptotic changes in the ovary or uterine endometrium, we studied the cleavage of DNA in these tissues obtained from regularly cycling women by in situ analysis of DNA integrity and quantitative end labeling of DNA gel fractionation. In situ analysis of several sized atretic follicles of ovaries revealed that granulosa cells showed positive staining to some extent, however, these methods do not discriminate between cells undergoing apoptosis and those undergoing necrosis. Total DNA extracted from human corpora lutea (CL) of the early luteal phase contained predominantly high molecular weight DNA, whereas CL of the midluteal phase exhibited the appearance of DNA cleavage into low molecular weight ladders characteristic of apoptosis. Although apoptotic DNA cleavage of human CL increased from the midluteal phase to the late luteal phase, CL of early pregnancy did not exhibit apoptotic DNA fragmentation. Both large and small luteal cells were the primary cell type exhibiting DNA cleavage in human CL of the midluteal and late luteal phases and in regressive CL. Biochemical analysis of human endometrium revealed that the ladder pattern cleavage of DNA was identified at three different phases of the menstrual cycle, namely the early proliferative, late secretory, and menstrual phases. Cells undergoing apoptosis were scattered in the functional layer of the early proliferative endometrium, but not in the late proliferative phase to midsecretory phase: At the beginning of the late secretory phase, apoptosis reappeared in the stromal cells and spread gradually to almost all components of the functional layer. By contrast, cells in the basal layer showed no evidence of apoptosis throughout the menstrual cycle. The present findings suggest that: (1) human luteal regression may be mediated by apoptosis; (2) CL of early pregnancy may be rescued from luteolysis through inhibition of the occurrence of apoptotic luteal cell death, and (3) apoptosis occurs in specific populations of endometrial cells during the human endometrial cycle. In conclusion, apoptosis might play an important role in the regulation of the menstrual cycle in women.

Production of inhibin A and inhibin B in human ovarian sex cord stromal tumors.

OBJECTIVE: Our purpose was to examine the cellular localization of inhibin subunits and messenger ribonucleic acid expressions for the inhibin subunits and the serum levels of inhibin A and inhibin B in human ovarian sex cord stromal tumors. STUDY DESIGN: We examined the immunohistochemical localization of the inhibin subunits and the expression of the corresponding messenger ribonucleic acids by Northern blot analysis in a granulosa cell tumor and a Sertoli-Leydig cell tumor. We also measured serum concentrations of dimeric inhibin A and inhibin B by two-site enzyme-linked immunosorbent assay. RESULTS: Immunostaining specific for the inhibin alpha, betaA, and betaB subunits was observed in the granulosa cell tumor. In the Sertoli-Leydig cell tumor we observed immunostaining specific for the alpha subunit in Leydig tumor cells and that specific for the betaA subunit in Sertoli tumor cells and that specific for the betaB subunit in both tumor cells. Northern blot analysis revealed the presence of messenger ribonucleic acids for the alpha, betaA, and betaB subunits in the granulosa cell tumor and the Sertoli-Leydig cell tumor. The serum levels of dimeric inhibin A and inhibin B in patients were elevated preoperatively and decreased progressively after surgery. CONCLUSION: Our results suggest that inhibin A and inhibin B are produced by the human sex cord stromal tumors and that inhibins might be the useful markers of the tumors.

Cytokine modulation of inhibin secretion in cultured rat granulosa cells.

In the present study, we examined the effects of tumour necrosis factor alpha (TNF alpha) and interleukin-1 beta (IL-1 beta) on inhibin secretion by cultured rat granulosa cells using immunoblotting and two-site enzyme immunoassay for inhibin A (alpha-beta A dimer). FSH stimulated the secretion of the inhibin alpha-beta A dimer (32 kDa) by the cells in a dose-dependent manner. In addition to the predominant 32 kDa inhibin alpha-beta A dimer staining, staining of minor immunoreactive bands was also enhanced by FSH. TNF alpha alone did not have any effect on inhibin secretion. Immunoblot analyses using an antiserum against alpha-subunit and an antiserum against beta A-subunit revealed a dose-dependent inhibition by TNF alpha of FSH-stimulated secretion of inhibin by rat granulosa cells. Similarly, TNF alpha inhibited in a dose-dependent manner FSH-stimulated inhibin secretion when measured using a two-site enzyme immunoassay. IL-1 beta alone did not exert any effect on inhibin secretion but it inhibited FSH-stimulated inhibin release in a dose-dependent manner (using both immunoblotting and a two-site assay for inhibin A). The present observations suggest that TNF alpha and IL-1 beta inhibit gonadotrophin-stimulated inhibin production by cultured rat granulosa cells.

Presence and localization of endothelin receptor in the rat ovary and its regulation by pituitary gonadotropins.

In the present study we examined the regulation of receptors for endothelin 1 (ET-1) in rat granulosa cells. We examined the localization and regulation of ET receptors in immature rat ovary and the effects of ET-1 on steroidogenesis in cultured rat granulosa cells. The ovaries used in autoradiography were derived from pregnant mare serum gonadotropin and human chorionic gonadotropin-treated immature rats. Granulosa cells were obtained from diethylstilbestrol-treated immature rats and incubated with 125I-ET-1. Granulosa cells were cultured with ET-1 in the presence or absence of ovine follicle-stimulating hormone. The concentrations of sex steroid hormones in conditioned media were measured by radioimmunoassay. The binding site for ET-1 was localized in the granulosa cells, but not in thecal and luteal cells. Follicle-stimulating hormone (FSH) induced a dose-dependent increase in specific binding for ET-1 to cultured rat granulosa cells. In contrast, luteinizing hormone (LH) induced a dose-dependent decrease in specific binding for ET-1 to cultured rat granulosa cells. Conversely, treatment with prolactin and several sex steroid hormones had no effects on the specific binding of ET-1. Treatment with ET-1 inhibited FSH-stimulated accumulation of progesterone and estradiol in cultured rat granulosa cells. The results indicate that both FSH and LH influence the expression of ET-1 receptor, and that ET-1 may play a regulatory role in the ontogeny of the granulosa cell.

Alterations in distribution and composition of the extracellular matrix during decidualization of the human endometrium.

The distribution of the extracellular matrix, including type I, III, IV and VI collagens and laminin, and of prolyl hydroxylase was investigated in the human endometrium by an indirect immunofluorescence method with specific monoclonal antibodies. Collagens were also extracted from the endometrial tissues in the proliferative and secretory phases and from the decidual tissues in the first trimester of pregnancy. Immunohistochemical studies demonstrated that interstitial collagens, such as type I, III, and VI collagens, were localized diffusely in the stroma of the endometrium throughout the menstrual cycle, as well as in the decidua. Type IV collagen and laminin were localized exclusively in the basement membrane of the endometrial glands and in the walls of blood vessels during the proliferative and secretory phases. However, strong staining for type IV collagen and laminin was recognized in the pericellular region of endometrial stromal cells in the decidua. Prolyl hydroxylase was localized in the cytoplasm of endometrial stromal cells and endometrial glandular cells during the menstrual cycle. Intense immunostaining for prolyl hydroxylase was observed in the decidual cells. However, immunoreactivity for prolyl hydroxylase in the endometrial glandular cells disappeared during the process of decidualization. The ratio of type III to type I collagen was significantly decreased (P < 0.05) and the ratio of type V to type I collagen was significantly increased (P < 0.01) in the decidua, as compared with ratios in the endometrium during the proliferative phase. The present results suggest that changes in the extracellular matrix may play an important role in implantation, in invasion of trophoblastic cells and in the maintenance of pregnancy.

Apoptosis of human corpora lutea during cyclic luteal regression and early pregnancy.

To elucidate the role of apoptotic cell death in human corpus luteum (CL) regression, human CL during the menstrual cycle and early pregnancy were isolated and processed for biochemical (radio-labeling) analysis of DNA integrity. Total DNA extracted from human CL of the early luteal phase contained predominantly high mol wt DNA, whereas CL of the midluteal phase exhibited the appearance of DNA cleavage into low mol wt ladders characteristic of apoptosis. Although apoptotic DNA cleavage of human CL significantly increased from the midluteal phase to the late luteal phase (P < 0.05), CL of early pregnancy did not exhibit apoptotic DNA fragmentation by biochemical analysis. In situ analysis of DNA fragmentation revealed that both large and small luteal cells exhibited DNA cleavage in human CL of the midluteal and late luteal phases and in regressive CL. The present findings suggest that 1) human luteal regression may be mediated by apoptosis; and 2) CL of early pregnancy may be rescued from luteolysis through inhibiting the occurrence of apoptotic luteal cell death.

[3H]naloxone binding sites in porcine ovarian follicles and corpora lutea during the ovarian cycle.

We demonstrated the presence of opioid receptors in the porcine ovary using [3H]naloxone. We also examined the change in the number of opioid receptors during follicular maturation. In addition, we found specific binding of [3H]naloxone in the porcine ovary using naloxone, beta-endorphin, methionine-enkephalin and dynorphin. The binding of [3H]naloxone to porcine granulosa cells and the 2000-g subcellular fraction of corpora lutea was examined to demonstrate the presence of specific [3H]naloxone binding in the porcine ovary. Binding of [3H]naloxone to porcine granulosa cells was displaced by cold naloxone and beta-endorphin but not by dynorphin and methionine-enkephalin. A similar phenomenon was also demonstrated in the 2000 g subcellular fraction of porcine corpora lutea. However, Scatchard analyses revealed a single class of high-affinity (Kd = 28.5 x 10(-9) mol/l) and low-capacity binding sites (Bmax = 30.5 fmol/5 x 10(6) cells) in porcine granulosa cells. Similar binding parameters were obtained in the 2000-g subcellular fraction of porcine luteal tissue (Kd = 28.3 x 10(-9) mol/l, Bmax = 59.3 nmol/kg protein). [3H]Naloxone binding sites in the porcine ovary showed binding characteristics similar to those of opioid receptors in other organs like brain, uterus and placenta. Furthermore, we demonstrated that the specific binding sites of [3H]naloxone in porcine granulosa cells decreased during follicular maturation. Opioid receptors have been detected in the uterus, placenta and Sertoli cell cultures in some species. However, there is no detailed study on opioid receptors in granulosa cells and luteal tissues in any species. Our data suggest a relationship between folliculogenesis and ovarian opioid peptides.(ABSTRACT TRUNCATED AT 250 WORDS)

Expression of luteinizing hormone and chorionic gonadotropin receptor messenger ribonucleic acid in human corpora lutea during menstrual cycle and pregnancy.

In the present study, we examined the expression of LH and CG receptor messenger RNA (mRNA) in human corpora lutea (CL) during the menstrual cycle and pregnancy. Poly(A)-enriched RNA was extracted from CL and analyzed by Northern and slot blots, using a radiolabeled complementary RNA probe derived from the human LH receptor complementary DNA. Northern blot analysis indicated the presence of multiple LH receptor mRNA transcripts with molecular sizes of 8.0, 7.0 and 4.5 kilobases in human CL during the menstrual cycle. The predominant transcript was 4.5 kilobases in size. However, no hybridization signals were observed in nongonadal tissues (heart, liver, and kidney). Densitometric analyses revealed that the levels of LH receptor mRNA increased from early luteal phase to midluteal phase and subsequently decreased during late luteal phase. After the onset of menstruation, the LH receptor mRNA level was undetectable in the regressing CL. Moreover, radioligand receptor assay (RRA) showed a close parallelism between LH receptor mRNA levels and LH receptor content in CL throughout the menstrual cycle. LH receptor mRNA expression was also found in CL during early pregnancy. The level of LH receptor mRNA was relatively high in early pregnancy CL, whereas LH receptor content was low. Using in situ hybridization, LH receptor mRNAs were uniformly expressed in both large and small luteal cells during early and midluteal phase and early pregnancy, but not in regressing CL. In conclusion, these data demonstrate that the regulation of LH receptor content in human CL during luteal phase is associated with similar changes in the receptor message levels, suggesting the physiological roles for LH receptor mRNA during the menstrual cycle in the human. In addition, the expression of LH receptor mRNA was demonstrated in human CL during early pregnancy.

Increased serum levels of type IV collagen and laminin in a patient with a giant retroperitoneal schwannoma.

A 63 year old woman presented with a giant abdominal tumor. Ultrasonography, computerized tomography and drop-infused pyelography revealed a suspected retroperitoneal tumor. The tumor was removed surgically and weighted 2800 g. The pathological diagnosis of the tumor was schwannoma. Strong immunohistochemical staining specific for type IV collagen and laminin was observed in the tumor, and these components were localized in the pericellular region of Schwann cells. The serum levels of these antigens, as determined by radio-immunoassay, were very high before the operation but decreased rapidly thereafter.