bromochloroacetic-acid and Polycystic-Ovary-Syndrome

Despite of the importance of cytoskeletal proteins for ovarian function and pathology, very few studies have addressed the presence and distribution of these proteins in polycystic ovaries. We investigated sections of human polycystic ovarian tissue for vimentin and a set of cytokeratins by epifluorescence. The studied proteins showed strong colocalization. Positive reaction was detected in two main ovarian compartments: with weak intensity in follicular cells and very strong in perinuclear position in oocytes of primordial follicles. Epifluorescent study of the oocytes from primordial follicles allowed us to identify the immunopositive structure in oocytes as Balbiani body, a transient organelle responsible for establishing oocyte polarity and ooplasm gradients in nonmammalian vertebrates. Our results suggest functional interaction of different types of cytoplasmic intermediate filament proteins in polycystic ovaries and a possible importance of the Balbiani body for human oogenesis in norm and pathology.

Topics: Female; Humans; Intermediate Filaments; Keratins; Ovarian Follicle; Ovary; Polycystic Ovary Syndrome; Vimentin

Polycystic ovary syndrome (PCOS) is associated with ovarian enlargement, prominent theca-interstitial hyperplasia, and excessive androgen production. Recent clinical trials have demonstrated that statins, 3-hydroxy-3-methyl-glutaryl-CoA reductase inhibitors, decrease androgen levels in women with PCOS.. The present study evaluated the effect of statins on proliferation of human ovarian theca-interstitial cells.. In vitro experiments were performed in the university research laboratory.. Human theca-interstitial cells were isolated from ovaries of PCOS (n=4) and non-PCOS (n=4) patients.. The cells were incubated for 48 h without additives (control) or with simvastatin (3-30 μm), mevastatin (3-30 μm), and/or the cell- and mitochondrion-permeable form of cholesterol (22-hydroxycholesterol; 10 μm). To determine whether the effects of statins could be affected by leukocytes, the experiment was carried out on cells not purified of leukocytes and cells purified using anti-CD-45 immunomagnetic beads. The effect of statins on proliferation was evaluated by determination of DNA synthesis using radiolabeled thymidine-incorporation assay and by quantification of viable cells using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenil)-2H-tetrazolium assay.. Statins induced an inhibition of DNA synthesis in both the absence and the presence of 22-hydroxycholesterol; furthermore, 22-hydroxycholesterol alone also inhibited DNA synthesis. These effects of statins and 22-hydroxycholesterol were confirmed by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenil)-2H-tetrazolium assay. Comparable inhibition of proliferation was observed in cells obtained from women with and without PCOS and in cell preparations treated and not treated with anti-CD-45 immunomagnetic beads.. Statins inhibit proliferation of human theca-interstitial cells irrespective of the availability of cholesterol and independently of leukocytes both in normal and PCOS ovaries.

Topics: Adult; Cell Division; Cell Survival; Cholesterol; Factor VIII; Female; Humans; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Keratins; Polycystic Ovary Syndrome; Premenopause; Simvastatin; Theca Cells; Thymidine; Vimentin

Numerous hypotheses have been proposed about the pathogenesis of the polycystic ovarian syndrome (PCOS). However, hormonal control of persistent follicles has not been established. The objective of the present study was to compare the follicular structure and hormonal profiles of rats treated with the adrenocorticotrophic hormone (ACTH) with two experimental models of PCOS. ACTH-treated animals were compared with those exposed to continuous light, those treated with estradiol valerate, and with control (in proestrous and diestrous). Serum hormone levels, histomorphometrical changes, and immunoexpression of vimentin, cytokeratins, cadherins, and proliferating cell nuclear antigen (PCNA) were examined. Treatment with ACTH resulted in an elevation of corticosterone secretion with LH reduction but without changes in ovarian morphology. Although stress (or ACTH) stimulation may be only one of pathophysiological mechanisms involved in follicular cyst pathogenesis in other species, we do not have important evidence to suppose that this would happen in rats.

Topics: Adrenocorticotropic Hormone; Animals; Cadherins; Cell Proliferation; Corticosterone; Estrous Cycle; Female; Gonadal Steroid Hormones; Hormones; Image Processing, Computer-Assisted; Immunohistochemistry; Keratins; Ovarian Follicle; Ovary; Polycystic Ovary Syndrome; Proliferating Cell Nuclear Antigen; Rats; Rats, Wistar; Vimentin

Our objective was to characterize the tissular distribution of relevant cytoskeletal proteins, cellular adhesion molecules and proliferation markers and conduct a histomorphometrical study of the follicular wall of letrozole-induced polycystic ovaries.. Twenty rats were divided into two groups: a control group (C) of ten rats that received vehicle only (0.9% NaCl solution) once daily p.o. and a treatment group (T) of ten animals administered letrozole at a concentration of 1 mg/kg p.o. dissolved in 0.9% NaCl solution once daily during 21 days. Twenty four h after the last administration, all animals were sacrificed. Control animals were sacrificed in proestrous (n = 5) and diestrous (n = 5). Serum hormone levels, histomorphometrical changes and immunoexpression of intermediate filaments (vimentin, cytokeratins and desmin), cadherins and proliferation cellular nuclear antigen were examined.. The granulosa cell layer of cystic follicles had a greater significant immunostaining for vimentin and cytokeratins. Immunohistochemical localization of desmin was restricted to the theca externa. Positive immunoreactivity for cadherins rises gradually and significantly, together with the follicular development, and immunoreactivity was comparatively stronger in follicular cysts. A significantly higher immunostaining for PCNA cells was observed in secondary and tertiary follicles as compared with atretic and cystic follicles. An increase in the LH, FSH and testosterone serum concentrations was observed in letrozole-treated rats. Estradiol and progesterone showed a considerable reduction.. The changes observed are probably due to structural and functional alterations that occur during the process of cystogenesis and may be associated with important modifications in the expression of cytoskeletal proteins, cellular adhesion molecules and proliferation markers that may be essential for proper cellular functioning.

Topics: Animals; Cadherins; Desmin; Female; Hormones; Immunohistochemistry; Keratins; Letrozole; Nitriles; Ovarian Follicle; Polycystic Ovary Syndrome; Proliferating Cell Nuclear Antigen; Rats; Rats, Wistar; Triazoles; Vimentin

Several experimental models have been developed for the study of the polycystic ovarian syndrome in the rat. In the present study, the syndrome was induced by exposure to constant light, and the expression of cytoskeletal proteins in the follicular wall was evaluated by immunohistochemistry. We analyzed the immunohistochemically stained area (IHCSA) by image analysis to evaluate the expression of intermediate filaments (vimentin, desmin, cytokeratins, gliofibrillary acidic protein and neurofilaments) and alpha-smooth muscle actin (alpha-SMA) in cystic ovaries in relation to normal ovaries. The granulosa cell layer of cystic follicles had a significantly greater IHCSA for vimentin than the normal antral follicles. This difference was also significant between atretic and antral follicles. Cytokeratins showed a very low expression in the granulosa cells of antral follicles of control ovaries while in granulosa cells of atretic and cystic follicles they showed a significantly higher IHCSA. Immunohistochemical localization of desmin and alpha-SMA was restricted to the theca externa. Immunoreactivity for gliofibrillary acidic protein and neurofilament was negative. The highest intensity in the staining with vimentin and cytokeratins observed in the granulosa cells of the cystic follicles is probably due to structural and functional changes that occur during the process of cystogenesis and they could be associated with intense changes in the expression of cytoskeletal proteins that may be essential to the proper cellular functioning.

Topics: Actins; Animals; Cytoskeletal Proteins; Desmin; Disease Models, Animal; Female; Image Processing, Computer-Assisted; Immunoenzyme Techniques; Keratins; Light; Ovarian Cysts; Ovarian Follicle; Ovary; Polycystic Ovary Syndrome; Rats; Rats, Wistar; Vimentin

This investigation was designed to study apoptosis and epithelialization during cystogenesis of the dehydroepiandrosterone rat model. Using in situ DNA 3'- end-labeling with non-radioactive digoxigenindidesoxy-UTP (dig-ddUTP), apoptosis is initially seen in cumulus granulosa cells and other granulosa cells facing the antrum. During cystogenesis, apoptosis systematically progresses from the cumulus towards the mural granulosa layer. In contrast, granulosa cells of atretic follicles undergo apoptosis in a random manner. The outer layer of mural granulosa cells during cystogenesis escapes apoptosis. Granulosa cells contain vimentin. However, the outer mural granulosa cell layer that lines the cyst acquires keratin. In addition to being associated with each other via gap junctions, the outer layer of granulosa cells acquire tight junctions. With the characterization of the transformation of the outer mural granulosa cells into a characteristic epithelium and the orderly progression of apoptosis, we further the understanding of the multifaceted process of cystogenesis of the ovarian antral follicle.

Topics: Androgens; Animals; Apoptosis; Cytoskeleton; Dehydroepiandrosterone; Disease Models, Animal; Epithelial Cells; Epithelium; Estradiol; Estrone; Female; Immunohistochemistry; Keratins; Microscopy, Electron; Ovarian Follicle; Polycystic Ovary Syndrome; Progesterone; Rats; Rats, Sprague-Dawley; Vimentin