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PubMed1979

Ovarian tumours in children--a review of 40 cases.

The records of 40 children treated for ovarian tumours in Manchester since 1940 are reviewed. The series includes 12 dysgerminomas, nine endodermal sinus tumours, six malignant teratomas, three granulosa theca cell tumours, one Sertoli-Leydig cell tumour, one mixed chorioncarcinoma/dysgerminoma and eight cases of non-germ cell or uncertain origin. Presenting features and treatment are discussed. Survival is generally poor apart from the dysgerminomas, granulosa theca and Sertoli-Leydig cell tumours, all but one of whom are alive and disease free. The need for routine post-operative radiotherapy in cases in dysgerminoma confined to one ovary following total resection is questioned. Combination chemotherapy may improve survival in the future, particularly of children with endodermal sinus tumour. The only surviving child other than those mentioned above has had combination chemotherapy for an endodermal sinus tumour.

Adolescent↗
PubMed1977

Intercellular junctions in the human developing preovulatory follicle and corpus luteum.

Using the lanthanum tracer and freeze-fracture techniques, the cell contacts of human ovarian cells were examined. The cell contacts between theca interna cells are similar to those seen between luteal cells are composed of septate-like cell contacts and gap junctions. On the other hand granulosa cells are connected by desmosomes and gap junctions. Thus, at least from the viewpoint of cell junctions, it is suggested that the main precursory cells of the luteal cells are the theca interna cells, although there is a general agreement that the luteal cells are derived from the granulosa cells.

Corpus Luteum↗
PubMed2003

Interleukin-8 gene and protein expression are up-regulated by interleukin-1beta in normal human ovarian cells and a granulosa tumor cell line.

OBJECTIVE: To evaluate the expression, regulation, and role of interleukin (IL)-8 in human ovary. DESIGN: Prospective study. SETTING: University hospital. PATIENT(S): Sixteen premenopausal women. INTERVENTION: Follicular fluid and granulosa lutein cells (GLCs) were collected during IVF cycles. Ovarian stromal and theca cells were obtained from women underwent surgery. KGN cells, the human granulosa cell tumor cell line, were also used. MAIN OUTCOME MEASURES: The levels of IL-8 and IL-1beta in follicular fluid and IL-8 protein production were determined using ELISA. Interleukin-8 and IL-8 receptor gene expression in ovarian cells and the effect of IL-8 on the proliferation of stromal cells were determined. The expression of pIkappaB was evaluated by Western blot, and the effect of NF-kappaB inhibitor APDC was examined by Northern blot analysis and ELISA in KGN cells. The levels of IL-8 and IL-1beta in follicular fluid; each concentration and the volume showed a positive correlation. Reverse transcription polymerase chain reaction showed the presence of IL-8 mRNA in all ovarian cells. In contrast, IL-8 receptor mRNA was only detected in stromal cells. The expression of IL-8 in GLCs and KGN cells was increased by addition of IL-1beta and TNFalpha. Interleukin-8 increased the proliferation of ovarian stromal cells. The expression of pIkappaB in KGN cells was induced by IL-1beta, and the effects were reduced by APDC. CONCLUSION(S): Interleukin 8 induced by IL-1beta via activation of NF-kappaB in granulosa cells may have a role in the periovulatory period of follicular maturation.

Blotting, Northern↗
PubMed2001

Muc13, a novel human cell surface mucin expressed by epithelial and hemopoietic cells.

Transmembrane mucins are glycoproteins involved in barrier function in epithelial tissues. To identify novel transmembrane mucin genes, we performed a tblastn search of the GenBanktrade mark EST data bases with a serine/threonine-rich search string, and a rodent gene expressed in bone marrow was identified. We determined the cDNA sequence of the human orthologue of this gene, MUC13, which localizes to chromosome band 3q13.3 and generates 3.2-kilobase pair transcripts encoding a 512-amino acid protein comprised of an N-terminal mucin repeat domain, three epidermal growth factor-like sequences, a SEA module, a transmembrane domain, and a cytoplasmic tail (GenBanktrade mark accession no. ). MUC13 mRNA is expressed most highly in the large intestine and trachea, and at moderate levels in the kidney, small intestine, appendix, and stomach. In situ hybridization in murine tissues revealed expression in intestinal epithelial and lymphoid cells. Immunohistochemistry demonstrated the human MUC13 protein on the apical membrane of both columnar and goblet cells in the gastrointestinal tract, as well as within goblet cell thecae, indicative of secretion in addition to presence on the cell surface. MUC13 is cleaved, and the beta-subunit containing the cytoplasmic tail undergoes homodimerization. Including MUC13, there are at least five cell surface mucins expressed in the gastrointestinal tract.

Amino Acid Sequence↗
PubMed1987

Immunoreactive beta-endorphin in ovarian sex cord-stromal tumors.

The possible production of the opioid polypeptide beta-endorphin (beta-EP) was investigated in paraffin-embedded tissue from 17 ovarian tumors with the use of a specific anti-beta-EP antibody and the avidin-biotin-peroxidase staining technique. Only sex cord-stromal tumors (ten cases) showed positive staining. Strong beta-EP immunoreactivity was present in Leydig's cells of Sertoli-Leydig cell tumors; weaker sporadic staining was present in cells of granulosa cell tumors, and faint staining was present in occasional, luteinized theca cells of fibrothecomata. These findings suggest that cells with the sex cord-stromal phenotype that are capable of steroid production can also produce beta-EP. The latter may be a component of the "functional" status associated with some ovarian sex cord-stromal tumors and may serve as a helpful marker in distinguishing this type of tumors from germ cell or epithelial neoplasms.

Cystadenocarcinoma↗
PubMed1997

Estrogen receptor protein and mRNA expression in the ovary of sheep.

Using immunohistochemistry and in situ hybridization, we attempted to identify the estrogen receptor (ER) protein and messenger RNA (mRNA) in sheep ovaries during the follicular phase of the estrous cycle. Monoclonal anti-ER antibodies H222 and 1D5 were used for localizing estrogen receptor on ovarian cryo-sections. Labeling for ER was found over the nuclei of surface epithelium, interstitial tissue, and granulosa cells of small as well as large ovarian follicles. In the preantral and small antral follicles, intense nuclear ER labeling was observed in mural granulosa cells and particularly in cumulus/granulosa cells surrounding the oocyte. In the large healthy looking follicles, greater diversity in labeling for ER was observed, which is characterized by mixed populations of granulosa cells expressing positive and more or less negative nuclear labeling. Such a pattern of labeling was particularly evident in follicles showing the signs of atresia. Generally, more intense nuclear staining was localized in granulosa cells proximal to basal membrane. In situ hybridization studies revealed the presence of ER mRNA in ovarian tissue. Autoradiographic visualization localized ER mRNA expression over the granulosa cells of healthy follicles of all sizes. Level of hybridization signal was comparable in mural and cumulus granulosa cells. In atretic follicles, the level of hybridization signal in granulosa cells was comparable to that of healthy follicles. A relatively weaker level of labeling was observed in granulosa cells dispersed in follicular antrum in follicles with advanced atretic lesions. Theca cells expressed a lower level of labeling than granulosa cells. Specificity of labeling for both ER protein and mRNA in ovary was proved by parallel probing the ovine uterus. Ovine ER recognition by both H222 and 1D5 antibodies was also proved by immunoblotting. These studies demonstrate the presence of the estrogen receptor and its messenger RNA in the sheep ovary and suggest an autocrine/paracrine role of estradiol and its receptor in the regulation of ovarian follicle development in sheep.

Animals↗
PubMed2004

Isolation of chicken homolog of the FOXL2 gene and comparison of its expression patterns with those of aromatase during ovarian development.

Mutations in the forkhead transcription factor gene FOXL2 are involved in ovarian failure, which occurs in human BPES syndrome. This syndrome presents a sexually dimorphic expression, specific to the ovary in several vertebrates. We cloned the open reading frame of chicken FOXL2 (cFoxL2) and studied cFoxL2 expression in developing gonads and during adulthood to examine the role of FOXL2 in ovarian differentiation and function in birds. The spatial and temporal dynamics of cFoxL2 and aromatase expression were analyzed in parallel by using real-time quantitative reverse transcriptase-polymerase chain reaction and immunohistochemistry in attempt to investigate the possible role of cFoxL2 in the regulation of aromatase. The expression patterns of cFoxL2 and aromatase transcripts were highly correlated during the sex-differentiation period (4.7-12.7 days of incubation). Aromatase and cFoxL2 proteins were colocalized in the medullar part of female gonads on embryonic day 14. Fourteen days after hatching, cFoxL2 protein was mainly detected in granulosa cells of developing follicles. In adult ovary follicular envelopes, apart from granulosa cells, cFoxL2 transcript and protein were detected at lower levels in theca cells where aromatase was present. A high level of cFoxL2 transcription was also observed in maturing and ovulated oocytes. Our results confirm that FoxL2 is an early regulator of ovarian development in birds and may be involved in aromatase transcription regulation.

Age Factors↗
PubMed1999

The endothelin system in human and monkey ovaries: in situ gene expression of the different components.

The endothelin system is composed of three endothelin isoforms (ET-1, ET-2, and ET-3), the endothelin receptors ETA and ETB, and the endothelin-converting enzyme (ECE). Besides having a major vasoactive role, endothelins have roles in different cell types at a local level. We investigated the presence of the different components of the endothelin system in primate ovaries. Human ovaries and gonadotropin-stimulated monkey ovaries were studied using immunohistochemistry for endothelin, and in situ hybridization with probes for ET-1, ET-2, ET-3, ETA and ETB receptors, and ECE. ET-1 and ETA receptors were detected in endothelial cells and vascular smooth muscle cells, respectively, in stromal vessels adjacent to follicles and corpora lutea. ETB receptors and ET-1 were found in the endothelial cells of capillaries of corpora lutea. ECE was present in internal theca cells of secondary, de Graaf, atretic follicles, and in luteinized granulosa cells of the corpora lutea. The endothelin system components are present in or around the follicles of human and monkey ovaries. Although the components are not expressed in the same cell types, they are synthesized, mainly in follicles, by cells that are in close proximity. Thus, the endothelin system could act in a paracrine manner. ECE expression in steroid-producing cells changes its compartmentalization during follicle maturation.

Animals↗
PubMed2001

Effect of adrenal and ovarian androgens on type 4 follicles unresponsive to FSH in immature mice.

The present study investigates the physiological significance of dehydroepiandrosterone, dehydroepiandrosterone sulfate, T, androstenedione (Delta(4)), dihydrotestosterone (DHT), estrone (E1), and E2 on recombinant human FSH- (rhFSH) resistant type 4 follicles obtained from immature mice. Type 4 follicles of a diameter of 100-120 microm with one or two granulosa cell layers around the oocyte and an intact basal lamina with theca cells were isolated from the ovaries of 11-d-old BDF-1 mice and cultured with medium alone (control) or with dehydroepiandrosterone, dehydroepiandrosterone sulfate, T, Delta(4), DHT, E1, or E2 at concentrations ranging from 1 x 10(-11) to 1 x 10(-7) M for 4 d. We examined the mean diameters of type 4 follicles, levels of immunoreactive (IR)-inhibin, and E2 and progesterone in the culture media on day 4. In addition, we evaluated follicular cell proliferation by immunofluorescence staining with 5-bromo-2'-deoxyuridine. All tested androgens significantly increased the diameter of type 4 follicles in a dose-dependent manner without the production of IR-inhibin and E2. The nuclei of granulosa cells in type 4 follicles cultured with all tested androgens exhibited intense 5-bromo-2'-deoxyuridine-positive staining, compared with those of controls. In contrast, neither E1 nor E2 had any stimulatory effects. The stimulatory effects of T, Delta(4), or DHT were inhibited by an AR antagonist in a dose-related fashion but not by an aromatase inhibitor. Furthermore, all tested androgens had a synergistic effect with rhFSH on follicular growth and the production of IR-inhibin and E2. These results demonstrated that neither adrenal nor ovarian androgens are arteriogenic but that they stimulate type 4 follicles unresponsive to rhFSH and augment the responsiveness of these follicles to rhFSH.

Adrenal Glands↗
PubMed2003

Identification and immunohistochemical localization of a haptoglobin-like protein in the tissues and fluids of the bovine (Bos taurus) ovary and oviduct.

The objective of this study was to establish the identity of a 40 kDa bovine oviductal fluid protein as a haptoglobin-like protein and to evaluate the association of the haptoglobin-like protein with ovarian and oviductal tissues and fluids. An oviductal fluid protein band corresponding to a molecular mass of 40 kDa was excised and electroeluted from SDS-PAGE gels. Sequence analysis revealed an N-terminal region sharing 81% identity with the beta-subunit of bovine haptoglobin. The 40 kDa oviductal fluid protein crossreacted on immunoblots with antiserum against rabbit endometrial haptoglobin and with an anti-human haptoglobin polyclonal antibody. Two-dimensional PAGE revealed four protein variants ranging in pI from 7.7 to 8.6, which appeared identical, with respect to molecular weight, number of isoforms and pI, to bovine haptoglobin in acute phase serum. The haptoglobin-like protein was localized using immunohistochemistry to the lumina of blood vessels and to the extracellular matrix of ovarian and oviductal tissues. Immunostaining for the haptoglobin-like protein was also detected in the oviductal lumen, in the mucosa of the ampullary oviduct but not the isthmic oviduct, and in intermittent ampullary epithelial cells. Within the ovary, the haptoglobin-like protein was localized to the avascular granulosa cells and follicular fluid of antral follicles, but not in the theca cells or in preantral follicles of any developmental stage. It was concluded that the haptoglobin-like protein is a normal constituent of bovine ovarian and oviductal tissues and fluids, and it was hypothesized that the haptoglobin-like protein contributes to ovarian follicular development and oviductal function.

Animals↗
PubMed1999

Distribution of ganglioside GM3 in the rat ovary after gonadotropin stimulation.

Gangliosides are ubiquitous membrane components in mammalian cells and are suggested to play important roles in various cell functions, such as cell-cell recognition, differentiation and transmembrane signalling. Ovaries have been shown to contain GM3 as a major ganglioside. To study GM3 distribution during gonadotropin stimulation in the hypophysectomized rat ovary, ovarian sections and cultured granulosa cells were stained with specific monoclonal antibody against GM3. Interstitial cells of follicles of immature hypophysectomized rat ovary expressed ganglioside GM3. Theca cells of early antral follicles but not primary follicles expressed GM3. No granulosa cells of these follicles expressed GM3. When a surge dose of FSH/LH was injected, Graafian follicles were formed and GM3 expression was detected in granulosa cells of these follicles. After ovulation, cumulus cells kept expressing GM3 in the ampulla region of ovulated oviduct. The follicles did not show GM3 expression in their granulosa cells after an ovulatory dose of FSH/LH. At 48 h after in vitro culture with FSH/LH of granulosa cells from preantral follicles, GM3 was expressed to a detectable extent on the outer part of the granulosa layer. Finally, at 72 h after culture, all granulosa cells became positive to anti-GM3 antibody. These data suggest that the expression of ganglioside GM3 in the hypophysectomized rat ovary is spatiotemporally regulated by FSH/LH during follicular development and ovulation.

Animals↗
PubMed2006

Gonadotropin regulation of testosterone production by primary cultured theca and granulosa cells of Atlantic croaker: I. Novel role of CaMKs and interactions between calcium- and adenylyl cyclase-dependent pathways.

Theca and granulosa cells for in vitro primary culture were obtained by enzymatic digestion of mature ovarian tissue from Atlantic croaker (Micropogonias undulatus) and separation from the other cell types by Percoll density-gradient centrifugation. Histochemical staining and treatment with pregnenolone confirmed the presence in the cultured cells of enzymes involved in synthesizing the major sex steroids in croaker ovaries: testosterone, estradiol, and 17alpha,20beta,21-trihydroxy-4-pregnen-3-one (20beta-S). Croaker theca and granulosa cells maintained their steroidogenic response to gonadotropin when cultured with serum-supplemented media and produced high levels of testosterone for up to 5 days, although estradiol production was low. Multiple signal transduction pathways mediating gonadotropin stimulation of androgen production were identified in Atlantic croaker ovarian theca and granulosa cells in primary co-culture. Inhibitors of voltage-sensitive calcium channels (VSCCs) and calmodulin decreased the steroidogenic response to gonadotropin, whereas activators of adenylyl cyclase and protein kinase A (PKA) increased testosterone production, indicating that both calcium and PKA-dependent signaling pathways are involved in the regulation of follicular steroid production. In addition, the first evidence in vertebrates for an involvement of calcium/calmodulin-dependent protein kinases (CaMKs) in gonadal steroidogenesis was obtained, since the stimulatory effects of gonadotropin on testosterone media accumulation were attenuated by specific inhibitors of CaMKs. Some interactions among the signaling pathways were observed as demonstrated by the positive effect of elevated intracellular calcium on adenylyl cyclase activity and the reduction of forskolin- and dbcAMP-induced testosterone production by inhibitors of VSCCs, calmodulin, and CaMKs.

3-Hydroxysteroid Dehydrogenases↗
PubMed2003

Regulation of the phenotype of ovarian somatic cells by estrogen.

The pathway of mammalian sex determination, and subsequent differentiation of the gonads is under the control of the sex-determining gene, Sry, on the Y chromosome. The presence of Sry leads to the formation of a testis with its complement of Sertoli and Leydig somatic cells. In the absence of Sry, an ovary develops with granulosa and theca cells. Ovarian development is said to initiate in the XX gonad as a default pathway because the XX cells do not express Sry. This review summarizes evidence supporting the view that the ovary is not entirely a default gonad. Studies of mice with deletions in both estrogen receptor (ERalphabetaKO) or aromatase (ArKO) genes have identified an important role of estrogens in maintaining differentiation and development of somatic cells in the ovary of eutherian mammals. In the absence of estrogen (ArKO) or the capacity to transduce an estrogen signal (ERalphabetaKO), the somatic cells in the ovary exhibited a male phenotype including Sertoli and Leydig cells. When ArKO mice were replaced with estrogen, the male phenotype was diminished and there was evidence of normal folliculogenesis in the ovary. It is concluded that the differentiation of somatic cells in the eutherian ovary is influenced by the sex steroid environment.

Animals↗
PubMed1999

Virilizing ovarian dermoid cyst with peripheral steroid cells. A case study with immunohistochemical study of steroidogenesis.

A case of virilizing ovarian dermoid cyst with peripheral steroid cells and virilization is reported in a 62-year-old woman. The level of testosterone dropped to normal after oophorectomy. The cyst wall was bordered by a discontinuous band of steroid cells focally accompanied by smooth muscle cells. Immunohistochemically, the steroid cells were enzymatically active and displayed a profile similar to the internal theca cells of ovarian follicles. These steroid cells were most probably modified stromal cells associated with smooth muscle metaplasia of the ovarian stroma.

DNA-Binding Proteins↗
PubMedDate not supplied

[Respective roles of thecal and granulosa cells in the morphogenesis of the corpus luteum].

Studies on human ovary embryogenesis and adult post-ovulatory surgically resected ovaries allowed the authors to propose a new conception on the genesis of corpus luteum: large luteal cells would be derived from theca interna cells and small luteal cells would come from externa theca cells. So there would be a structural unity of luteal tissue which might explain steroidogenesis potential of these two types of cells, derived both from the same ovarian stroma. On the other hand if granulosa cells played an important function in bio-conversion of androgens to oestradiol by aromatase activity before ovulation, their role is over after the expulsion of the oocyte out of the follicle; derived from primitive sex cords of coelomic mesothelium origin, granulosa cells act to support and surround the oocyte; they disappeared at the disappearance of the oocyte from the follicle.

Corpus Luteum↗
PubMed1995

Immunocytochemical analysis of the expression of gap junction protein connexin 43 in the rat ovary.

The present immunocytochemical study examines in the rat ovary the pattern of expression of connexin 43 (Cx43), a subunit of gap junctions. Using a well-characterized specific antiserum against rat Cx43, immunoreactivity was not detected in the fetal ovary, i.e., prior to follicular formation. However, in the ovary of 20-day-old, 35-day-old, and adult rats, strong Cx43-immunoreactivity was associated with the cell borders of the follicular epithelium/granulosa cells of all developmental stages (primordial follicles, preantral and antral secondary follicles). In general, immunoreactivity of the granulosa cells of large antral follicles appeared more intense than the one of smaller follicles. Staining was also seen in oocytes (cytoplasmic staining). Theca cells of large antral follicles, but not of small follicles were immunoreactive. Immunoreactive interstitial cells were not seen in ovaries of 20- and 35-day-old animals, but staining in these cells was present in adult rats. In large follicles with signs of atresia, granulosa cells lacked Cx43-immunoreactivity, whereas Cx43-immunoreactivity in their theca interna strikingly increased. Corpora lutea in the cyclic adult rats were heterogeneously stained, with either no detectable immunoreactivity, staining of cell borders of most luteal cells, or with conspicuous staining of only a few cells. In the pregnant animals on gestation days (GD) 12, 14, and 17, all luteal cells stained strongly for Cx43 at the cell surface.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗
PubMed2006

Localization of angiotensin II, the AT1 receptor, angiotensin-converting enzyme, aminopeptidase A, adipocyte-derived leucine aminopeptidase, and vascular endothelial growth factor in the human ovary throughout the menstrual cycle.

OBJECTIVE: To assess the expression and cellular distribution of angiotensin II (Ang II), angiotensin type 1 receptor (AT1R), angiotensin-converting enzyme (ACE), aminopeptidase A (APA), adipocyte-derived leucine aminopeptidase (A-LAP), and vascular endothelial growth factor (VEGF) in human ovarian tissue during the menstrual cycle. DESIGN: Ovarian tissues (n = 52) and corpora lutea (n = 34) were obtained from patients undergoing hysterectomy/oophorectomy, and tissue sections were immunostained for each antigen. SETTING: University hospital. PATIENT(S): Patients undergoing hysterectomy or oophorectomy for benign conditions. INTERVENTION(S): Immunostaining of tissue sections using antibodies to each antigen. MAIN OUTCOME MEASURE(S): Microscopic evaluation to assess the presence, distribution, and cellular localization. RESULT(S): The luteal tissue is the major site of Ang II, ACE, AT1R, and VEGF, with highest staining intensity found during the midluteal phase and at pregnancy. The AT1R was found in theca cells. The APA was strongly immunolocalized in pericytes. Immunolocalization of AT1R was almost similar to that of VEGF including oocytes in the primordial and intermediate follicles. CONCLUSION(S): The expression and distinct pattern of the cellular localization of Ang II and its related proteins in human ovarian tissue during folliculogenesis and in the luteal tissue suggest their roles in the growth and differentiation of theca, granulose, and luteal cells.

Aminopeptidases↗
PubMed1993

Dynamic changes in ovarian c-kit and Steel expression during the estrous reproductive cycle.

W and Steel mutant mice exhibit similar developmental defects in melanogenesis, haematopoiesis, and gametogenesis. Consistent with the cell autonomous and microenvironmental nature of W and Sl mutations, respectively, W encodes the c-kit receptor tyrosine kinase while Steel encodes the Kit ligand. Both c-kit and Steel are expressed in various cells in which no corresponding mutant phenotype has yet been demonstrated. In the adult ovary, certain stromal-derived cells (theca and interstitial), as well as oocytes, express c-kit, while granulosa cells express Steel. We show here that the cessation of oocyte growth, at the transition of the follicle to the antral stage, is associated with the cessation of Steel expression in the cumulus granulosa cells in the vicinity of the oocyte. These observations suggest a role for the Kit signaling pathway in oocyte growth or in meiotic arrest. In addition, the cyclic secretion of luteinizing hormone immediately and dramatically results in elevated Steel expression in mural granulosa cells and decreased levels of c-kit transcripts in stromal-derived cells. This influence of the estrous reproductive cycle on c-kit/Steel expression suggests that the Kit signaling pathway, in addition to its previously described role in primordial germ cell development, is involved in follicular development in the adult female.

Animals↗