Effect of cis-dichlorodiammineplatinum(II) on cloned ovarian adenocarcinoma cells of the rat in vitro

Cloned rat ovarian adenocarcinoma cells (ROT58/C3) treated for 1 hr with 0.5 (μg/ml or more of cis-dichlorodiammineplatinum(II) (DDP) or Adriamycin (ADM) revealed a marked decrease in survival as estimated by the colony-forming method. Compared to cloned rat uterine adenocarcinoma cells (HTP/C1), the ROT58/C3 cells were more sensitive to the cytotoxic effect of both drugs. The survival of exponentially growing ROT58/C3 cells was reduced to less than 10% by a 0.5 μg/ml dose of DDP, as compared to a 1 μg/ml dose of ADM. Extensive morphological changes such as enlargement of the nuclear and nucleolar diameters, increase in the number of nucleoli, and the appearance of coarse and opaque chromatins were observed by optical and electron microscopy at as early as 48 hr after exposure to DDP for 1 hr. In addition, accumulation of cells at the G₂ phase in the cell cycle was observed by flow microfluorometry.     

Regulation of interleukin-8 synthesis in human lower uterine segment fibroblasts by cytokines and growth factors

OBJECTIVE: To investigate the influence of lipopolysaccharide, cytokines, growth factors, and progesterone on the synthesis of interleukin-8 by human lower uterine segment fibroblasts. METHODS: Fibroblasts derived from a lower uterine segment biopsy specimen obtained from a woman undergoing elective cesarean delivery at term were exposed to lipopolysaccharide, interleukin-1beta, transforming growth factor-beta(1), platelet-derived growth factor-AB, and combinations of these substances. All experiments were performed in the absence and presence of progesterone. The concentration of interleukin-8 in the culture medium was determined by enzyme immunoassay after 24 hours. RESULTS: Compared with controls (0.71 +/- 0.04 ng interleukin-8/10(6) cells), fibroblasts exposed to lipopolysaccharide, transforming growth factor-beta(1), or platelet-derived growth factor-AB exhibited no increase, or at most, only a minor but significant increase, in interleukin-8 secretion. Incubation with interleukin-1beta led to a moderate increase, whereas the combinations interleukin-1beta/transforming growth factor-beta(1) (105.0 +/- 7.5 ng interleukin-8/10(6) cells) and interleukin-1beta/platelet-derived growth factor-AB (387.3 +/- 25.6 ng interleukin-8/10(6) cells) increased interleukin-8 secretion dramatically. No further increase was observed with the combination interleukin-1beta/platelet-derived growth factor-AB/transforming growth factor-beta(1). When progesterone was added, interleukin-8 secretion decreased significantly by 16-34%, depending on the stimulator, or did not change. CONCLUSION: The findings indicate that interleukin-8 secretion by human lower uterine segment fibroblasts in vitro is upregulated by interleukin-1beta, transforming growth factor-beta(1), and platelet-derived growth factor-AB in a synergistic fashion. Because interleukin-8 mediates the invasion of neutrophils into the cervical stroma, this may be an important mechanism controlling cervical dilatation during parturition.     

Localization of sex steroid hormone and sex steroid hormone receptors in human ovarian epithelial tumor

In some ovarian tumors, such as endometrioid adenocarcinoma, dysfunctional uterine bleeding occurs, and the endocrinological aspects were studied in the following way: 1) In 3 patients with epithelial ovarian tumor and postmenopausal uterine bleeding, preoperative plasma estradiol (E2) and progesterone (P) concentrations were significantly high, but dropped to normal following complete resections of the ovarian tumor. Plasma E2, P and testosterone concentrations in ovarian veins were 3 to 7 times as great as in peripheral veins. 2) Tissues obtained from 25 ovarian tumors were immunohistochemically examined by the PAP method. In 9 cases of endometrioid adenocarcinoma, positive stainings for both E2 and P were demonstrated in cancer cells from 3 patients (33%) and in connective tissues from 7 patients (78%), respectively. In contrast, 16 tumors of other histological types gave only one positive staining for E2 on cancer cells and connective tissues. As for P, in 3 patients there was a positive stain on cancer cells and in 2 patients on connective tissues. 3) Five out of 8 patients with immunohistochemically positive staining for E2 on connective tissues in ovarian tumors demonstrated dysfunctional uterine bleeding. 4) Cytosolic estrogen receptor (ER) and progesterone receptor (PR) in ovarian tumors obtained from 26 patients were measured by the DCC method. All of three patients with endometrioid adenocarcinoma had both ER and PR. The former were 19-202 fmol/mg, and the latter 535-1,000 fmol/mg, which were significantly higher levels and positive rates than those from other patients.(ABSTRACT TRUNCATED AT 250 WORDS)     

Interleukin-6 (IL-6) production in carcinoma of the cervix

Interleukin-6 (IL-6) is a pleiotropic cytokine that is not only a mediator in major immunologic reactions but also a growth factor of keratinocytes. We studied the IL-6 secretion in vitro of 15 human cell lines derived from both squamous cell carcinoma (SCC) and adenocarcinoma of the uterine cervix. Four of the eight well differentiated SCC secreted a large amount (> 1500 pg/48 h/10⁶ cells) of IL-6 in nude mice. In contrast, poorly differentiated SCC cell lines and all of the 7 adenocarcinoma cell lines secreted a small amount (< 500 pg/48 h/10⁶ cells of IL-6). The expression of IL-6 mRNA of the cell lines correlated well with their IL-6 secretion potential. However, the expression of IL-6 receptor did not correlate with the IL-6 secretory potential. We also studied the IL-6 secretion of freshly isolated normal squamous epithelium and of dysplastic epithelium. In culture, two normal squamous epithelia secreted a large amount (> 2000 pg/48 h/10⁶ cells), whereas 8 dysplasia epithelia secreted an extremely small amount (< 10 pg/48 h/10⁶ cells). About one-third of patients with SCC had a raised serum IL-6 value. IL-6 production may help to differentiate between SCC and adenocarcinoma of the uterine cervix. IL-6 regulation seems to change in the course of SCC carcinogenesis.     

Endocrine aspects of human uterine sarcoma: a preliminary study

The biologic effect of estrogen and progesterone in human uterine sarcoma is poorly understood in comparison to that of endometrial adenocarcinoma. In an attempt to elucidate the endocrine status of these tumors, we have investigated the ability of these tumors to synthesize estrogen by measuring the aromatase activity and studied the effect of aromatase inhibitors on the activity. In addition, the effect of estrogen and progesterone on aromatase activity and the growth pattern of these tumors were studied in cell culture and athymic mice systems. Aromatase activities in eight uterine sarcomas ranged from 0.7 to 37 fmol/hr X mg protein, which were within the range or higher than the activity found in normal proliferative endometrium (0.5 to 3 fmol/hr X mg of protein, means = 1.6, n = 10). These results indicate that uterine sarcomas are capable of producing estrogen. However, the enzyme activity showed no correlation with the morphology of tumors or the age of patients. Results from the kinetic studies of aromatase activity in one of the uterine sarcomas indicated that 19-nortestosterone, testolactone, and aminoglutethimide (the most effective one) inhibited aromatase activity. In addition, induction of aromatase activity in two uterine sarcomas was investigated in cell cultures. Progesterone caused an eightfold increase in activity in a sarcoma that was estrogen and progesterone receptor positive but had no effect in a tumor that was estrogen and progesterone receptor negative. The growth rate of two estrogen/progesterone receptor-negative uterine sarcomas was studied in cell culture and in athymic mice. Progestin, but not estrogen, reduced the growth rate in both systems; 30 nmol/L of estrogen had no effect on the growth rate. In summary, we have found that human uterine sarcoma is able to synthesize estrogen. Progesterone is able to induce the aromatase activity in estrogen/progesterone receptor-positive tumors, and progesterone also suppresses the tumor growth rate in estrogen/progesterone receptor-negative tumors. These results suggest that a select group of uterine sarcomas is sensitive to steroid hormone and that progesterone may be potentially beneficial for therapeutic treatment of select uterine sarcomas.     

Interleukin-1beta and Interleukin-8 Concentrations in the Lower Uterine Segment During Parturition at Term

Objective: To assess the roles of interleukin-1β, interleukin-8, and fibroblasts in the lower uterine segment during parturition.Methods: Lower uterine segment biopsy specimens were obtained from 36 women undergoing cesarean delivery at various stages of cervical dilation (less than 2 cm, n = 8; 2 to less than 4 cm, n = 9; 4–6 cm, n = 10; more than 6 cm, n = 9). The concentrations of interleukin-1β and interleukin-8 in protein extracts prepared from the tissue samples were measured by enzyme immunoassays. The effect of incubation with interleukin-1β (30 U/mL) on interleukin-8 secretion by lower uterine segment fibroblasts in vitro also was determined.Results: The median interleukin-1β concentration in the specimens increased from 1.3 pg/mg of total protein at less than 2 cm of dilation to 22.2 pg/mg of total protein at 4–6 cm of dilation (P < .05). No further increase was detectable after 6 cm of dilation. The interleukin-8 concentration increased from 17.2 pg/mg of total protein at less than 2 cm of dilation to 2080.7 pg/mg of total protein at 4–6 cm of dilation (P < .05), thus paralleling the increase in interleukin-1β concentration. Interleukin-1β induced a significant increase in interleukin-8 secretion by fibroblasts in vitro, from 0.8 ng/10⁶ cells to 35.6 ng/10⁶ cells.Conclusion: The increase in interleukin-8 concentration in the lower uterine segment during parturition may be induced by interleukin-1β and fibroblasts may be one of the sources of this interleukin-8.     

Danazol binds to progesterone receptors and inhibits the growth of human endometrial cancer cells in vitro

Based on our recent findings that danazol, an isoxazol derivative of ethinyltestosterone, has a profound growth-inhibitory effect on an established human endometrial adenocarcinoma cell line, the effects of danazol on cancer cells from human endometrial adenocarcinomas obtained by hysterectomy were investigated in the present study. Of the 22 uterine adenocarcinomas, estrogen, progesterone, and androgen receptors were found in 12, 14, and 4 tumors, respectively. Competitive binding studies showed that danazol specifically binds to progesterone and androgen receptors but not to estrogen receptors. Of the five cancer cells from five patients succeeded in primary cell culture, a marked inhibition of cell growth was demonstrated by addition of danazol in two cancer cells having progesterone but not androgen receptors. However, danazol did not affect the growth of the remaining three cancer cells lacking progesterone receptors. These results strongly suggest that danazol has a significant growth-inhibitory effect on human endometrial adenocarcinoma cells, possibly through progesterone receptors in the cells.     

Effect of androgen substrates on the steroidogenic pattern of cumulus cells: Correlation with cumulus culture morphology

Background: In previous studies, higher progesterone secretion was observed in mature versus immature cumulusoocyte complexes. In mature cumulus mass that become homogeneously spread in culture (type C/D) progesterone secretion was higher than in partially (type B) or totally (type A) aggregated morphology. In sharp contrast, estradiol-17β secretion was significantly higher in type A than type C/D cumulus. Purpose: Our purpose was to assess whether the decreased estradiol-17β level in type C/D cumulus culture is caused by deficiency of substrates. Methods: The different cumulus types were incubated with or without 10⁻⁷ M dehydroepiandrosterone, 4-androstane-3, 17-dione, or testosterone. The levels of estradiol-17β, testosterone, and progesterone, were measured after 24 hr of culture. Results: The addition of dehydroepiandrosterone or 4-androstane-3,17-dione significantly increased the estradiol-17β levels in all types of cumulus cells, whereas the addition of testosterone was less effective. In all types of cumulus cells the testosterone levels increased significantly on adding these androgen substrates. In the type C/D cumulus, the testosterone increased to lower levels compared to type A cumulus cells. In the presence of these androgens progesterone secretion is significantly reduced in type A cumulus cells. In type C/D cumulus cells, however, progesterone levels were significantly higher than in type A. The estradiol-17β/testosterone and progesterone/estradiol-17β ratios, which partially resemble the degree of aromatase activity and the degree of selectivity for progesterone secretion, respectively, were higher in type C/D than in type A cumulus cells. Conclusions: In type C/D cumulus the significant increase in estradiol-17β secretion in the presence of various androgens suggests that, under basal conditions, androgen is less available for estradiol-17β biosynthesis compared to type A cumulus. Furthermore, the higher progesterone secretion in type C/D cumulus may suggest that the follicles yielding type C/D cumulus cells are more mature than the follicles yielding type A cumulus.     

米非司酮和孕酮对子宫内膜异位症患者子宫内膜白细胞介素6分泌的影响

目的探讨米非司酮和孕酮对子宫内膜异位症(内异症)患者异位子宫内膜(异位内膜)与正常子宫内膜(在位内膜)白细胞介素6(IL-6)分泌的影响.方法采用酶联免疫吸附试验(ELISA)方法,检测米非司酮浓度为1×10-6mol/L(米非司酮1组)、1×10-4mol/L(米非司酮2组),和孕酮浓度为1×10-7mol/L(孕酮1组)、1×10-5mol/L(孕酮2组)与体外培养的异位内膜细胞和在位内膜细胞上清液作用后的IL-6水平.结果米非司酮可降低异位内膜细胞IL-6水平,米非司酮1组和米非司酮2组的IL-6分为(1914.33±799.28)μg/L和(990.25±58.40)μg/L,两组与空白组比较(下同),差异有极显著性(P＜0.01).孕酮也可降低异位内膜细胞IL-6水平,孕酮1组和孕酮2组的IL-6分为(2575.89±119.75)μg/L和(1736.25±750.89)μg/L(P＜0.01).米非司酮还可使在位内膜细胞IL-6水平降低,其中米非司酮1组和米非司酮2组分别为(346.96±24.32)μg/L和(270.22±36.15)μg/L(P＜0.01).孕酮对在位内膜细胞IL-6水平无明显影响(P＞0.05).结论米非司酮和孕酮可抑制异位内膜和在位内膜细胞IL-6的分泌.     

The Urokinase Plasminogen Activator (uPA) System in Uterine Natural Killer Cells in the Placental Bed During Early Pregnancy

The urokinase plasminogen activator (uPA) system plays pivotal roles in cell invasion, adhesion and migration. Roles for uterine natural killer (uNK) cells in regulating extravillous trophoblast (EVT) invasion and spiral artery remodeling have been proposed. Placental bed biopsies from early pregnancy were obtained from three gestational age groups (8–10, 12–14 and 15–20 weeks). Total caseinase activity in the placental bed was studied using casein in situ zymography. Localisation of uPA, uPA receptor (uPAR), plasminogen activator inhibitor (PAI)-1 and -2 in the placental bed was investigated by immunohistochemistry. CD56⁺ uNK cells were separated from collagenase-digested decidual cells using an immunomagnetic technique, and uPA activity was measured in isolated cell culture supernatants by casein/plasminogen gel zymography (8–10 and 12–14 weeks' gestation, n = 10 each group). uPAR in cell lysates and PAI-1 and -2 secretion in supernatants were measured by Western blotting. Caseinase activity was stronger in decidua than myometrium as shown by in situ zymography. uPA localised strongly to uNK cells, especially at 8–10 weeks. Moderate uPAR localisation on uNK cells also observed. There was very weak immunostaining of uNK cells for PAI-1 and PAI-2. In casein gel zymography, uPA activity was similar in uNK cell culture supernatant compared with total unseparated decidual cells. uPAR in uNK cell lysates was significantly stronger than in total decidual cell lysates. PAI-1 and PAI-2 were not detected in uNK cell culture supernatants by Western blot analysis. These results suggest that uNK cells may regulate EVT invasion and spiral artery remodeling via the uPA system.     

Influence of thyroxine on human granulosa cell steroidogenesis in vitro

Purpose The objective of this study was to investigate the influence of thyroid hormone on estradiol and progesterone secretion of human granulosa cells maintained in vitro.Methods Granulosa cells were obtained by aspiration of preovulatory follicles of woman undergoing assisted reproductive technology. Ovulation induction was performed with GnRH agonist, hMG, and hCG.Results Granulosa cells were maintained in vitro in a defined medium with added insulin. Twenty-four-hour estradiol and progesterone secretion into the medium were determined for granulosa cells growing in serum-free medium and in serum-free medium with added T₄ in a concentration range of 10^–7 to 10^–11 M.Conclusions All concentrations of T₄ used produced a statistically significant increase in progesterone secretion (range, 1.39 to 1.60 times the baseline amount). The increase in estradiol secretion reached statistical significance only at a T₄ concentration of 10^–8 M (1.24 times the baseline amount).     

Development of vitrified-warmed mouse embryos co-cultured with polarized or non-polarized uterine epithelial cells using sequential culture media

PURPOSE: To our knowledge, little is known about the effect of polarized and non-polarized uterine epithelial cells on cryopreserved embryo growth. This study was, therefore, set up to investigate the effect of these monolayers together with sequential culture media on vitrified-warmed mouse embryos in terms of blastocyst development, blastocyst quality, incidence of apoptosis and related genes expression. METHODS: Two cell vitrified-warmed mouse embryos were cultured in G-1ver3 medium to the eight-cell stage when they were randomly assigned to three treatment groups of no co-culture (control), non-polarized and polarized mouse uterine epithelial monolayer co-culture. The culture medium was G-2ver3 during the treatment phase. After 96 h on treatment, the significance of differences were evaluated by the one way analysis of variance for continuous data. RESULTS: In the polarized monolayer group, the hatched blastocyst formation and blastocyst quality improved significantly than other two groups (P < 0.05). Whereas the incidence of apoptosis and related gene expression such as Bax were higher in the blastocysts of control group (P < 0.05). The relative abundance of Bcl-2 mRNA to the beta-tubulin was similar for all treatments. CONCLUSION: Co-culture system involving polarized uterine epithelial cells and sequential culture media is a promising method for the improvement of vitrified-warmed mouse embryo development.     

Suppression of human chorionic gonadotropin by progestational steroids

The dynamics of the secretion of human chorionic gonadotropin (hCG) were studied by culturing explants of normal term placentas for as long as 144 hours. A significant accumulation of immunoreactive hCG (beta-subunit) was first detected at 48 to 72 hours, and a sixfold increase in hCG was observed in control culture medium at 144 hours. Compared to control cultures, progesterone (P < 0.001) in physiologic tissue levels of 5 to 20 micrograms/ml, pregnenolone (P < 0.001), 20 micrograms/ml, and 20 alpha-dihydroprogesterone (P < 0.001), 20 micrograms/ml, suppressed the secretion of hCG throughout the study period. Progesterone decreased the secretion of hCG in a dose-response manner (r = -0.8S87, P < 0.1). No suppression of hCG was observed in the presence of cortisol, testosterone, dihydrotestosterone, 17 beta-estradiol, or estriol. The secretion of human chorionic somatomammotropin was unchanged in the presence of progesterone. The augmented hCG response in the presence of dibutyl cAMP (P < 0.001) was significantly, but not completely, suppressed by progesterone 20 micrograms/ml culture medium (P < 0.01). Under these conditions progestational steroids or their immediate metabolites suppress the secretion of hCG, and they may be responsible for the decline in the levels of hCG during pregnancy.     

Inhibitory effects of danazol on steroidogenesis in cultured human granulosa cells

The effect of danazol on steroidogenesis in cultured human granulosa cells was studied. Granulosa cells obtained from antral follicles in the mid to late follicular phase were cultured for 2 to 6 days. Danazol (0.1 to 5 micrograms/ml) and human chorionic gonadotropin (hCG, 10 IU/ml), alone or in combination, were added to the culture medium. Testosterone (T) (1 microgram/ml) was added as an aromatase substrate in certain experiments. The medium content of progesterone (P) and estradiol (E2) was determined. Basal P secretion was variably influenced by danazol, whereas a consistent dose-dependent and reversible inhibition of the hCG-stimulated P secretion was found. In the presence of T, danazol caused a dose-dependent inhibition of both basal and hCG-stimulated E2 secretion, and this effect became more pronounced with time. The results demonstrate that danazol exerts direct inhibitory effects on steroidogenesis in cultured human granulosa cells.     

子宫颈癌组织中水通道蛋白8和bcl-2蛋白的表达及其相关性

目的 探讨水通道蛋白(AQP)8、bcl-2蛋白在宫颈癌组织中的表达及其相关性.方法 采用免疫组化Envision二步法检测AQP8和bcl-2蛋白在74例宫颈癌(其中鳞癌46例、腺癌28例)、34例宫颈上皮内瘤变(CIN)和15例正常宫颈组织中的表达情况,并分析两者的相关性.结果 AQP8和bcl-2蛋白主要在CIN异型细胞和宫颈癌细胞的细胞质内表达,AQP8蛋白在宫颈鳞癌、腺癌、CIN和正常宫颈组织中的阳性表达率分别为98%、61%、71%和53%,鳞癌高于腺癌、CIN和正常宫颈组织,差异有统计学意义(P＜0.01);腺癌与CIN、正常宫颈组织比较,CIN与正常宫颈组织比较,差异均无统计学意义(P＞0.05).bcl-2蛋白在宫颈鳞癌、腺癌、CIN和正常宫颈组织中的阳性表达率分别为74%、71%、53%和20%,鳞癌与腺癌组织比较,差异无统计学意义(P＞0.05);鳞癌、腺癌高于CIN、正常宫颈组织,CIN也高于正常宫颈组织,差异均有统计学意义(P＜0.01).AQP8和bcl-2蛋白在宫颈癌组织中的表达呈明显正相关(rs=0.463,P=0.000).结论 AQP8和bcl-2蛋白在宫颈癌组织中的表达呈明显正相关,AQP8蛋白表达上调可能与宫颈癌的发生、发展有一定的关系.     

Synuclein-γ (SNCG) protein expression is associated with poor outcome in endometrial adenocarcinoma

Objective

Synuclein-γ (SNCG) is a marker for adverse and aggressive disease in breast cancer. In previous study, we found SNCG mRNA to be overexpressed in uterine serous carcinoma compared to uterine endometrioid adenocarcinoma. The aim of this study is to explore the prognostic value of SNCG in patients with endometrial cancer.

Methods

279 endometrial cancer patients were retrieved from the archives. The tissue paraffin blocks were stained for SNCG antibody and its expression was correlated with clinicopathological prognostic factors.

Results

There was a positive association between SNCG⁺ immunoexpression and tumor grade, tumor stage, type II carcinomas, deep myometrial invasion and lymphovascular invasion. A correlation between SNCG⁺ and adverse outcomes, such as shorter overall survival (OS) and disease free survival (DFS), was also detected. Following adjuvant therapy (radiation and chemotherapy or chemotherapy alone), we observed a difference in 5 years DFS rate between SNCG⁺ (41.6%) and SNCG⁻ patients (59.5%).

Conclusion

Overexpression of SNCG seemed to be a predictor biomarker for aggressive tumor behavior and adverse outcome in patients with endometrial cancer. Future exploration of SNCG as a potential therapeutic target for selected patients could be of interest.     

Carriage of the methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism does not influence the first and second trimester uterine artery Doppler flow

Objective

MTHFR C677T polymorphism is a genetic factor increasing both risk factors for atherosclerotic vascular diseases and obstetric complications like preeclampsia (PE) and fetal growth restriction (FGR). Increased uterine artery impedance, measured by uterine artery Doppler in the second trimester of pregnancy is also associated with PE and FGR. In this study we aimed to analyze whether MTHFR influences first and second trimester uterine artery impedance.

Study design

In a prospective, controlled, open, single center study of 1955 consecutive singleton pregnant women, smears from buccal gingival cells were analyzed for MTHFR by hybridisation on micro arrays. Uterine artery PI values and unilateral or bilateral diastolic notch were measured at 12 and 22 weeks of gestation. Statistical significance was calculated by the x²-test.

Results

MTHFR C677T polymorphism showed a normal distribution in our population. Mean uterine artery Doppler values and bilateral or unilateral notch occurrences from 1697 statistical evaluated women did not show significant differences in any MTHFR genotype (C/C, C/T, T/T) both at 12 or 22 weeks of gestation.

Conclusion

In summary, the data presented in this adequately powered, prospective, controlled study establish that the MTHFR C677T polymorphism does not influence Doppler flow measurements.     

The effects of adrenergic and cholinergic agents on progesterone production by human corpus luteum in vitro

We investigated the effects of adrenergic and cholinergic agents on human corpus luteum production of progesterone in vitro. Luteinizing hormone (LH) (50 ng/ml), dibutyryl cyclic adenosine monophosphate (Bu₂cAMP) (10^?3M), and prostaglandin E₂ (PGE₂) (1 μg/ml) significantly stimulated the production of progesterone in short-term (4-hour) cell suspensions of five early and middle luteal phase corpora lutea. The adrenergic agents isoproterenol, norepinephrine, and the cholinergic agents acetylcholine and carbachol at concentrations up to 10^?4M did not alter basal or stimulated production of progesterone. Similarly, in long-term (10-day) monolayer cultures of cells from four corpora lutea, human chorionic gonadotropin (hCG) (50 ng/ml) and PGE₂ stimulated, but none of the adrenergic or cholinergic agents altered, the production of progesterone significantly, except for an inhibitory effect of norepinephrine and carbachol in the presence of 17β-estradiol (10^?7M) added to the culture medium. These results differ strikingly from the consistent stimulatory effect of β-adrenergic agents on the luteal production of progesterone in several animal species.