transforming-growth-factor-beta and Ovarian-Cysts

The process of transformation of growing bovine follicles into cysts is still a mystery. Local expression of proteins or factors, including transforming growth factor β, growth factors, and transcription factors, plays a central role in mammals. Therefore, in abattoir-derived cystic ovarian follicles and follicular fluid, the role of some transforming growth factor β superfamily proteins, insulinlike growth factor-1 (IGF-1) and GATA-4 and GATA-6, were investigated. The relationship between intrafollicular lipopolysaccharide (LPS) and etiopathogenesis of ovarian cysts was also assessed. Data on the preovulatory follicle and the largest follicle (F1) were compared. The number of intrafollicular LPS-positive samples and LPS concentrations were higher in cysts. Immunohistochemical staining was mildly positive for IGF-1, inhibin alpha, and GATA-4 in thecal cells. Staining for anti-Müllerian hormone (AMH), growth differentiation factor-9, bone morphogenetic protein-6 (BMP-6), and GATA-6 was insufficient for their quantitation, and oocytes could not be stained for any of the proteins tested in the cystic follicles. Expression of BMP-6, inhibin alpha, and IGF-1 was moderately higher in granulosa cells of F1 follicles, and all the proteins were moderately expressed in granulosa cells in preovulatory follicles. However, loss of GATA-6 staining was significant in F1 follicles. Intrafollicular progesterone, IGF-1, and AMH concentrations in cysts and F1 follicles were significantly higher than those in preovulatory follicles. Western blot analyses revealed that follicular fluid inhibin-α was strongly expressed, whereas expression of growth differentiation factor-9, BMP-6, GATA-4 and GATA-6 was lower in cysts than in preovulatory follicles. Also, high intrafollicular AMH concentration and low BMP-6 expression were closely associated with cystic degeneration and atresia. In conclusion, immunohistochemical loss of BMP-6 and GATA-6 in the granulosa cells together with high intrafollicular LPS levels may play important roles in disruption of the ovulatory mechanism and steroidogenic reactions in type 2 cyst. Also, high intrafollicular AMH concentration along with low BMP-6 expression may be used as indicators of the bovine degenarative ovarian follicles.

Topics: Animals; Anti-Mullerian Hormone; Bone Morphogenetic Protein 6; Cattle; Cattle Diseases; Female; Follicular Fluid; GATA4 Transcription Factor; GATA6 Transcription Factor; Granulosa Cells; Growth Differentiation Factor 9; Immunohistochemistry; Insulin-Like Growth Factor I; Lipopolysaccharides; Ovarian Cysts; Ovarian Follicle; Progesterone; Transcription Factors; Transforming Growth Factor beta

Cystic ovarian disease (COD) is one of the main causes of infertility in dairy cattle. It has been shown that intra-ovarian factors may contribute to follicular persistence. Transforming growth factor-beta (TGFB) isoforms are important paracrine and autocrine signalling molecules that regulate ovarian follicle growth and physiology. Considering the importance of these factors in the ovarian physiology, in this study, we examined the expression of TGFB isoforms (TGFB1, TGFB2 and TGFB3) in the ovary of healthy cows and animals with spontaneous and adrenocorticotrophic hormone (ACTH)-induced COD. In the oestrous-synchronized control group, the expression of TGFB1 in granulosa and theca cells was higher in spontaneous cysts than in atretic or tertiary follicles. When we compared TGFB2 expression in granulosa cells from atretic or tertiary follicles from the oestrous-synchronized control group with that in ACTH-induced or spontaneous follicular cysts, we found a higher expression in the latter. The expression of the TGFB isoforms studied was also altered during folliculogenesis in both the spontaneous and ACTH-induced COD groups. As it has been previously shown that TGFB influences steroidogenesis, ovarian follicular proliferation and apoptosis, an alteration in its expression may contribute to the pathogenesis of this disease.

Topics: Adrenocorticotropic Hormone; Animals; Cattle; Cattle Diseases; Female; Gene Expression Regulation; Ovarian Cysts; Ovariectomy; Protein Isoforms; Transforming Growth Factor beta

Chronic ovulation as a contributing factor for the development of epithelial ovarian cancer in women has long been an outstanding hypothesis. To test the incessant ovulation hypothesis, mice were superovulated using weekly ip injections of pregnant mare serum gonadotropin (5 IU/animal), followed 48 h later by human chorionic gonadotropin (5 IU/animal). Wild-type CD1 mice were used along with CD1 mice expressing a Smad2 dominant-negative (Smad2DN) transgene under the control of the Müllerian inhibiting substance promoter that targets expression to the ovary and enhances cyst formation. After chronic injections, ovaries were analyzed from animals 6 months of age for the total adjusted number of cysts, cyst area, cyst location, and key signaling pathways. All observed cysts were confirmed to be of epithelial origin. The number of cysts was not significantly different between superovulated and control mice in either the wild-type or Smad2DN groups. However, the combination of the Smad2DN transgene and superovulation resulted in an increase in cyst formation compared with normal littermates that were unstimulated. Rapid proliferation of the cells lining the cysts was detected using bromodeoxyuridine and phospho-histone 3 immunohistochemistry but was not different in the ovarian surface epithelium or in the cyst lining between groups. These data suggest that chronic superovulation in Smad2DN mice results in a higher incidence of cyst formation compared with unstimulated controls, but the epithelial lined cysts did not progress to cancer over the course of this study.

Topics: Activins; Animals; Cell Division; Chronic Disease; Epithelial Cells; Female; Gonadotropins, Equine; Mice; Mice, Inbred Strains; Mice, Transgenic; Ovarian Cysts; Ovarian Neoplasms; Ovulation; Phosphorylation; Signal Transduction; Smad2 Protein; Superovulation; Transforming Growth Factor beta

Increased concentrations of TGF-beta 1 in endometriotic tissue are considered important in the pathophysiology of endometrial diseases since TGF-beta 1 may inhibit natural killer activity and induce angiogenesis and proliferation of endometrial stromal cells. In the present study we report on TGF-beta 1, IGF-1 and their receptor localization, as detected by Northern hybridization and immunohistochemistry, in ovarian endometriotic tissues removed during surgical procedures. We detected comparable expression of IGF-1 and IGF-1 receptor in the stromal and epithelial compartments, thus confirming disregulated expression of the IGF system in ovarian endometriosis. On the contrary, strongly increased TGF-beta 1 steady state level mRNA expression was detected in all endometriotic samples. In addition, we demonstrated weak TGF-beta 1 immunohistochemical expression in the epithelial lining and intense expression in the cellular stroma of ovarian endometriomas, thus suggesting that TGF-beta 1 could have an important role in the maintenance and propagation of the disease. On the basis of these preliminary results we can assume that TGF-beta 1, IGF-1 and their receptors may play an important role in the pathogenesis of endometriosis.

Topics: Adult; Blotting, Northern; Endometriosis; Endometrium; Female; Humans; Immunohistochemistry; Insulin-Like Growth Factor I; Macrophages; Ovarian Cysts; Pilot Projects; Receptor, IGF Type 1; Receptors, Transforming Growth Factor beta; RNA, Neoplasm; Stromal Cells; Transforming Growth Factor beta; Transforming Growth Factor beta1

The present study examined the presence and cellular distribution of transforming growth factor-beta1, 2, and 3 isoforms and their type I and II receptors in endometriotic cysts of the ovary, relative to their presence in normal endometrial tissue.. Thirteen control samples of normal endometrium in the proliferative phase and 11 ovarian endometriotic cysts were examined by immunohistochemistry for transforming growth factor-beta1, 2, and 3 isoforms and their type I and II receptors.. Immunoreactivity for all ligands and receptors was detected in both normal endometrium and endometriotic cysts. Isoform-specific differences in immunostaining were not detected. Expression of all ligands and receptors was significantly increased in epithelial cells of endometriotic cysts compared with those of normal endometrium. On the other hand, stromal cells in normal endometrium and endometriotic cysts were only faintly immunostained. Inflammatory cells infiltrating among endometriotic stromal cells contained the highest immunostaining intensity for all ligands and receptors. We identified nearly all inflammatory cells as macrophages using a specific antibody.. These findings suggest that macrophages in endometriotic tissue are a major source of transforming growth factor-beta, which may be an important regulator of cell proliferation in endometriotic cysts through paracrine and autocrine actions.

Topics: Activin Receptors, Type I; Adult; Endometriosis; Endometrium; Female; Humans; Immunohistochemistry; Macrophages; Ovarian Cysts; Protein Isoforms; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type I; Receptor, Transforming Growth Factor-beta Type II; Receptors, Transforming Growth Factor beta; Transforming Growth Factor beta

To evaluate the effect of chocolate cyst fluid on the proliferation of cultured human endometrioma cells and to assay the concentration of transforming growth factor-B1 in this fluid.. Controlled in vitro study.. Department of Obstetrics and Gynecology, State University of New York Health Science Center.. Five women with ovarian endometriomas.. Endometrioma tissue and chocolate fluid from five different patients were entered in this study.. Endometrioma cell proliferation in culture with and without chocolate cyst fluid.. Chocolate cyst fluid increased the proliferation of endometrioma cells compared with controls. Also, high concentrations of transforming growth factor-B1 were found in cysts' fluid.. Chocolate cyst fluid has a growth-enhancing effect on endometrioma cells. One promoting growth factor is transforming growth factor-B1.

Topics: Cell Count; Cell Division; Cells, Cultured; Cyst Fluid; Endometriosis; Enzyme-Linked Immunosorbent Assay; Female; Humans; Ovarian Cysts; Transforming Growth Factor beta