Within the corpus luteum, macrophages exert luteotropic and luteolytic actions through secretion of TNF-alpha. However, the mechanisms of luteotropic actions on the development and maintenance of pregnant and nonpregnant corpora lutea are thoroughly unknown.In this experiment, TUNEL, macrophage, and TNF-alpha immunohistochemistry on the corpora lutea of pregnant and nonpregnant rats (Sprague-Dawley strain) were carried out to reveal the role of macrophages in the developing corpora lutea. The results were as follows; 1) In the nonpregnant corpora lutea, the number of macrophages was increased significantly, and the degree of ED1-immunoreactivity of macrophages was increased moderately. But lutein cells showed low-degree TNF-alpha-immunoreactivity. 2) In the pregnant corpora lutea, the number of macrophages was decreased significantly, and the degree of ED1- immunoreactivity of macrophages was low. But lutein cells showed moderate-degree TNF-alpha-immunoreactivity. Based on the above results, it was considered that macrophages in the nonpregnant corpora lutea exert phagocytic action mainly, and the macrophages in the pregnant corpora lutea exert TNF-alpha-secreting action to maintain the structure and function of lutein cells.
Animals ; Corpus Luteum* ; Female ; Immunohistochemistry ; In Situ Nick-End Labeling ; Luteal Cells ; Macrophages* ; Rats* ; Tumor Necrosis Factor-alpha

To examine the electrophysiological properties of luteal cells and the relationship between membrane potential and luteal steroidogenic capacity, the membrane potential of luteal cells and the luteal steroidogenesis were measured under different ionic conditions following treatment with various drugs and gonadotropins. The membrane potential of luteal cells did not vary throughout the estrous cycle and was -55 +/- 1 mV. The membrane potential was highly dependent upon the external K+ concentration and was depolarized by the deprivation of external Ca2+, however) there seemed to be a lower K+ permeability in luteal membranes as the presence of 10-9 M valinomycin, a K+ ionophore Caused hyperpolarization from -55 to -91 mV. Luteal progestin production was increased in a high K+ solution but not m a Ca2+-free solution indicating that Ca2+ may be essential for steroid synthesis and/or secretion by luteal cells. Gonadotropins and ouabain induced a depolarization of the membrane potential and stimulated luteal steroidogenesis; however; prostaglandin F2alpha stimulated only steroidogenesis without any changes in membrane potential. These results suggest that the relationship between steroidogenesis and the changes in membrane potential by drugs and gonadotropins is still obscure and remains to be eluridated. The relationship between membrane potential and steroidogenesis in the luteal cell may be dependent upon the availability of intracelluar Ca2+.
Animal ; Corpus Luteum/*metabolism ; Estrus/metabolism ; Female ; Ions ; Luteal Cells/*metabolism ; Membrane Potentials ; Rats ; Rats, Inbred Strains ; Steroids/*biosynthesis ; Support, Non-U.S. Gov't

Macrophages in the corpus luteum have many important roles during the periods of functional development and luteal regression. Not only phagocyte the apoptotic luteal cells, but also they secrete many cytokines and exert their effects via autocrine/paracrine actions.In this study, we investigated the changes of number and immunoreactivity of macrophages at various developmental periods of the corpus luteum in the rat ovary. The rats (Sprague-Dawley strain, female)at age of 8 weeks (ovulatory period), GD 6 (pregnant period), and postpartum 5 days (postpartum period)were sacrificed under ether anesthesia and obtained both ovaries, one used for macrophages immunohistochemistry and the other used for TEM. The results were as follows; 1.In the corpora lutea of the rat, macrophages were observed all the developmental periods including ovulatory, pregnant and postpartum periods. 2.In the corpora lutea of the rat, number of macrophages was highest in the ovulatory period, and decreased at postpartum period and pregnant period in order. The immunoreactivity of macrophages was high at ovulatory period, moderate at postpartum period, and low at pregnant period. 3.In TEM observations, two types of macrophages were observed: One type was non-phagocytic macrophage and the other type was phagocytic macrophage. Phagocytic macrophages were observed in the corpora lutea at ovulatory and postpartum periods and contained apoptotic bodies, phagocytic vacuoles and many lipid droplets. Non-phagocytic macrophages were observed in the corpora lutea at pregnancy period and showed slender cell body with long cytoplasmic processes and contained no apoptotic bodies. In the rat, the number and the degree of immunoreactivity of macrophages in the corpus luteum varied with the changes of functional state of the corpus luteum. It was suggested that the main function of the macrophages at the ovulatory and postpartum periods was elimination of apoptotic luteal cells and that at pregnancy period was autocrine/paracrine action.Ultrastructurally, two types, phagocytic and nonphagocytic types, of macrophages confirmed. These results will provide valuable informations on the study of the role macrophages during development and regression of corpus luteum.
Anesthesia ; Animals ; Apoptosis ; Corpus Luteum* ; Cytokines ; Cytoplasm ; Ether ; Female ; Immunohistochemistry ; Luteal Cells ; Luteolysis ; Macrophages* ; Ovary ; Phagocytes ; Postpartum Period ; Pregnancy ; Rats* ; Vacuoles

Primary ovarian pregnancy is a rare form of ectopic pregnancy, and the prevalence rate is reported to be between 1/7,000 and 1/40,000 pregnancies. Ovarian pegnancy occurs within the ovary and on the corpus luteum, usually with rupture and massive bleeding. It is frequently misdiagnosed as a ruptured corpus luteum and the differentiation may be made only by micoroscopic examination of a tissue specimen. We have experienced four cases of ovarian pegnancy, which are presented with a brief review in the literature.
Corpus Luteum ; Female ; Hemorrhage ; Ovary ; Pregnancy ; Pregnancy, Ectopic* ; Prevalence ; Rupture

Ovarian pregnancy is comparatively rare form of ectopic pregnancy. Although earlier diagnosis is now possible due to the availability of quantitative beta-hCG measurement and the development of transvaginal ultrasonograghy, it is mostly difficult to diagnosis before surgery, and frequently misdiagnosed as a ruptured corpus luteum accompanied with massive hemoperitoneum. Definite diagnosis is made only by cytopathologic examination of tissue specimen. The treatment of ovarian pregnancy has been operative management including oophorectomy, salpingo-oophorectomy and ovarian wedge resection. But recently conservative management using laparoscopic technique has become the preferred treatment. We have experienced a case of primary ovarian pregnancy and reviewed it briefly.
Corpus Luteum ; Diagnosis ; Female ; Hemoperitoneum ; Ovariectomy ; Pregnancy ; Pregnancy, Ectopic*

Ovarian pregnancy is a rare form of ectopic pregnancy that is often difficult to diagnose. The diagnostic criteria were described in 1878 by Spiegelberg, which comprise that the pregnancy is in the ovary and does not involve the tube. Ovarian pregnancy occurs in the corpus luteum, and is usually accompanied with the rupture of the ovary and massive hemoperitoneum. It presents as a hemorrhagic ovary and frequently misdiagnosed as a ruptured corpus luteum. We have experienced a case of ovarian pregnancy and reviewed it briefly.
Corpus Luteum ; Female ; Hemoperitoneum ; Ovary ; Pregnancy ; Pregnancy, Ectopic* ; Rupture

OBJECTIVE: Our object is to evaluate the detailed mechanisms of support and regression of the human corpus luteum. METHODS: To investigate the regulation of luteal function by gonadotropins, cytokines, and prostaglandins, the frequency of apoptosis and expression of Fas, Fas-L, Bcl-2, Bax, p53, caspase-8 were examined in cultured human luteal cells after treatment with various doses of FSH (30, 100, or 300 ng/mL), LH (30, 100, or 300 ng/mL), TGFbeta1 (1, 10, or 100 ng/mL), TNFalpha (1, 10, or 100 ng/mL), or PGF2alpha (1, 10, or 100 ng/mL) for 24 h. Cells were tested for apoptosis by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-digoxigenin nick end labeling TUNEL) method and cell death detection ELISA. Immunostaning was performed using anti-Fas, Fas ligand, Bcl-2, Bax, and p53 antibodies. RESULTS: Incidence of apoptosis determined by TUNEL method in the group without treatment was 1.7+/-0.5% (0 h), 10.8+/-1.6% (24 h), and 12.9+/-1.2% (48 h), respectively. Spontaneous increase was significant at the latter time points. Significant suppression of incidence of apoptosis was observed with LH and TGFbeta1 (P<0.05). On the other hand, significant induction of incidence of apoptosis was observed with TNFalpha and PGF2alpha (P<0.05). Immunostaining revealed that p53 and Bax expressions after treatment with LH or TGFbeta1 were significantly lower than those without treatment. Bcl-2 and caspase-8 expressions were not significantly affected by any substance addition. Also we found that inductions of apoptosis by TNFalpha and PGF2alpha were not correlated with the expression of Fas, Fas ligand, Bcl-2, Bax, p53 and caspase-8. CONCLUSION: Our results suggest that LH and TGFbeta1 may be involved in the support of luteal function via suppression of apoptosis, and that TNFalpha and PGF2alpha may contribute to luteal regression via its induction in human corpus luteum during early luteal phase. Also, Fas, Fas-L, Bax and p53 may play roles in this apoptosis controlled by LH, and TGFbeta1.
Antibodies ; Apoptosis* ; Caspase 8 ; Cell Death ; Corpus Luteum ; Cytokines* ; Deoxyuridine ; Dinoprost ; Enzyme-Linked Immunosorbent Assay ; Fas Ligand Protein ; Female ; Gonadotropins* ; Hand ; Humans* ; In Situ Nick-End Labeling ; Incidence ; Luteal Cells* ; Luteal Phase ; Luteolysis ; Prostaglandins ; Tumor Necrosis Factor-alpha

This study was undertaken to investigate the characterization and distribution patterns of MHC class II positive dendritic cells[DCs] and ED2 positive tissue macrophages throughout the estrous cycle and during pregnancy in the rat ovary. The immunohistochemical characterization of the cells was carried out using the monoclonal antibodies OX6 and ED2 in cryostat-cut sections. DCs were distributed in the theca cell layer of the growing and mature follicles,stroma and corpus luteum. Tissue macrophages were distributed in the theca externa of the growing and mature follicles, stroma and corpus luteum but they were smaller in number than DCs. None of DC and tissue macrophage was found in the ovum, granulosa layer and follicular cavity of the ovarian follicle. However, DCs and tissue macrophages were present in the granulosa layer and follicular cavity in the atretic follicles. Degenerating corpus luteum contained a vast number of OX6 positive cells. On the contrary, fewer tissue macrophages were founcl in the degenerating corpus luteum. More macrophages tended to be observed in the former follicular cavity and theca lutein cell layer than in the granulosa lutein cell layer of the corpus luteum. In stroma,DCs and tissue macrophages were more frequently found around the blood vessels than in the other region, however, DCs were relatively greater in number than tissue macrophages. There was no estrous cycle and pregnancy dependent variation in the numbers and distribution patterns of DCs and tissue macrophages. In conclusion, the rat ovary contains rich networks of MHC class II positive dendritic cells and ED2 positive tissue macrophages. These findings suggest the existence of a well-developed system of immunological surveillance in the rat ovary. The results of this study have potentially important implications for the understanding not only of the ovarian immune system and the pathogenesis of various ovarian diseases but also of various physiologic functions of the ovary.
Animals ; Antibodies, Monoclonal ; Blood Vessels ; Corpus Luteum ; Dendritic Cells* ; Estrous Cycle ; Female ; Immune System ; Immunologic Surveillance ; Luteal Cells ; Macrophages* ; Ovarian Diseases ; Ovarian Follicle ; Ovary* ; Ovum ; Pregnancy ; Rats* ; Theca Cells

OBJECTIVE: This study was aimed to obtain information on normal MIS serum levels according to menstrual cycles of adult normal cycling women . It was also designed to obtain information on the ontogeny of the production profile of MIS and the pattern of its localization in ovary from adult normal cycling women. METHODS: Between January 1998 and January 1999, normal MIS serum levels were measured according to menstrual cycles using 160 serum samples from adult normal cycling women by ELISA. The ontogeny of the production profile of MIS and the pattern of its localization were also studied by immunohistochemical staining using the rabbit polyclonal antibody against human recombinant MIS in 35 ovarian specimens from adult normal cycling women. RESULT: The MIS levels were gradually increased through the follicular phase, reaching at its maximum at the ovulatory phase(4.2+/-2.6 ng/ml), and sharply decreased at the beginning of the luteal phase being minimized at the premenstrual phase(0.5+/-0.2 ng/ml). In average, the MIS levels of the follicular phase(3.7+/-1.9 ng/ml) were significantly higher than those of the luteal phase(1.8+/-2.4 ng/ml)(P<0.05). The MIS levels of the preovulatory and ovulatory phase were significantly higher than those of the other cycle days(P<0.05). Even the early follicular phase(2.9+/-1.6 ng/ml) showed higher MIS levels than the advanced luteal phase(0.9+/-0.7 ng/ml) and the premenstrual phase(0.5+/-0.2 ng/ml)(P<0.05 and P<0.05, respectively). The first staining for MIS was detected in the cytoplasm of granulosa cells when the flattened granulosa cells changed to the cuboidal cells in primordial follicles. The granulosa cells of both single and multiple layered growing follicles showed strong specific staining for MIS. but the MIS staining was not found not in the mature follicle just before ovulation, atretic follicles, corpus luteum, and corpus albicans. MIS staining waned in the mature follicles just before ovulation. CONCLUSION: These experiments demonstrate that the MIS is produced by ovarian granulosa cells in normal reproductive females. The MIS may play an important role as a hormone of follicular development and oocyte maturation through interactions with female steroid hormones, gonadotropins, and growth factors during the adult reproductive cycle.
Adult ; Anti-Mullerian Hormone* ; Corpus Luteum ; Cytoplasm ; Enzyme-Linked Immunosorbent Assay ; Female ; Follicular Phase ; Gonadotropins ; Granulosa Cells ; Humans* ; Intercellular Signaling Peptides and Proteins ; Luteal Phase ; Menstrual Cycle* ; Oocytes ; Ovarian Follicle ; Ovary* ; Ovulation

The present study aimed to investigate the role of platelet-activating factor (PAF) in progesterone synthesis and vascular endothelial growth factor (VEGF) expression in rat luteal cells. Immature (25-28 days old) female Sprague-Dawley rats were injected subcutaneously with 50 IU pregnant mare serum gonadotrophin (PMSG), and 25 IU human chorionic gonadotrophin (hCG) 48 h later, to induce follicular development and luteum formation. On day 6 after hCG administration (the day of hCG administration was the first day), the rats were killed by guillotine and the ovarian luteal cells were collected. After incubation for 24 h, luteal cells were incubated without or with different doses (0.1 μg/mL, 1 μg/mL, 10 μg/mL) of PAF at 37 °C (5% CO(2)) for 24 h, and then progesterone concentration was evaluated by radioimmunoassay (RIA); apoptotic rate and VEGF mRNA expression in luteal cells were assessed by flow cytometry and RT-PCR, respectively. The results showed that PAF promoted progesterone production, with a maximal effect at 1 μg/mL (P<0.05); PAF increased apoptotic rate but not in a dose-dependent manner, and 10 μg/mL PAF enhanced apoptotic rate significantly (P<0.05); furthermore, PAF stimulated VEGF mRNA expression in luteal cells, especially at 1 μg/mL (P<0.01). It is suggested that PAF regulates progesterone synthesis and VEGF mRNA expression in luteal cells to mediate corpus luteum formation in rat ovary.
Animals ; Chorionic Gonadotropin ; pharmacology ; Corpus Luteum ; drug effects ; Female ; Luteal Cells ; drug effects ; metabolism ; Platelet Activating Factor ; pharmacology ; Pregnancy ; Progesterone ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; metabolism