Normal Ovarian Structures Research Articles

Ameliorating effects of adropin on letrozole-induced polycystic ovary syndrome via regulating steroidogenesis and the microbiota inflammatory axis in rats.

Growing evidence supports the role of gut microbiota in chronic inflammation, insulin resistance (IR) and sex hormone production in polycystic ovary syndrome (PCOS). Adropin plays a pivotal role in the regulation of glucose and lipid metabolism and is negatively correlated with IR, which affects intestinal microbiota and sex hormones. However, the effect of adropin administration in PCOS has yet to be investigated. The present study aimed to assess the effects of adropin on letrozole (LTZ)-induced PCOS in rats and the potential underlying mechanisms. The experimental groups were normal, adropin, letrozole and LTZ+adropin. At the end of the experiment, adropin significantly ameliorated PCOS, as evidenced by restoring the normal ovarian structure, decreasing the theca cell thickness in antral follicles, as well as serum testosterone and luteinizing hormone levels and luteinizing hormone/follicle-stimulating hormone ratios, at the same time as increasing granulosa cell thickness in antral follicles, oestradiol and follicle-stimulating hormone levels. The ameliorating effect could be attributed to its effect on sex hormone-binding globulin, key steroidogenic genes STAR and CYP11A1, IR, lipid profile, gut microbiota metabolites-brain-ovary axis components (short chain fatty acids, free fatty acid receptor 3 and peptide YY), intestinal permeability marker (zonulin and tight junction protein claudin-1), lipopolysaccharides/Toll-like receptor 4/nuclear factor kappa B inflammatory pathway and oxidative stress makers (malondialdehyde and total antioxidant capacity). In conclusion, adropin has a promising therapeutic effect on PCOS by regulating steroidogenesis, IR, lipid profile, the gut microbiota inflammatory axis and redox homeostasis. KEY POINTS: Adropin treatment reversed endocrine and ovarian morphology disorders in polycystic ovary syndrome (PCOS). Adropin regulated the ovarian steroidogenesis and sex hormone-binding globulin in PCOS. Adropin improved lipid profile and decreased insulin resistance in PCOS. Adropin modulated the components of the gut-brain-ovary axis (short chain fatty acids, free fatty acid receptor 3 and peptide YY) in PCOS. Adropin improved intestinal barrier integrity, suppressed of lipopolysaccharides/Toll-like receptor 4/nuclear factor kappa B signalling pathway and oxidative stress in PCOS.

Protective Effect of Coscinium fenestratum Bark Extract on Letrozole Induced Polycystic Ovary Syndrome (PCOS) in Sprague Dawley Rats

Aim: Polycystic ovary syndrome (PCOS) is a common endocrine disorder. The aim of the present study is to evaluate the effects of the oral administration of Coscinium fenestratum bark extract in letrozole-induced PCOS in rats.
 Methods: Thirty female Sprague-Dawley (SD) rats were randomly divided into five groups of 6 animals each. Letrozole (1 mg/kg) was administered orally for a period of 21 days to induce PCOS. Control rats received 1 mL/day of distilled water, and the three groups of PCOS rats were given 1mg/kg of clomiphene citrate, 250 mg/kg of Coscinium fenestratum extract and 500 mg/kg of Coscinium fenestratum extract by oral gavage for 15 days. Blood samples were collected and the ovary of rats was removed, weighed, and fixed in 10% buffered formalin to determine the hormonal and antioxidant parameters.
 Results: In rats treated with BECF, significant increase in Catalase, Superoxide Dismutase and Lipid peroxidation level was decreased when compared with letrozole group. Histopathology showed that the BECF treated rats had normal ovarian structure, which was seen distorted due to the PCOS condition in letrozole treated rats. The results show significant decrease in total testosterone level and LH and significant increase in FSH. Therefore, the extract of BECF show significant effect on PCOS rats. The significance p value is >0.001.
 Conclusion: Based on these result, the BECF extract and traditional wisdom of this plant when used regularly can be a good treatment a suitable treatment option for PCOS which can be regularly used.

Effects of bilateral whole vessel ovarian ligation on dogs’ ovarian function and histopathology

The most significant postoperative complication of spaying is bleeding from ovarian pedicles. In this study, all of the ovarian vessels were bilaterally ligated; then, ovaries were stimulated by gonadotropins. In a clinical trial and sequential control experiment, five healthy mixed-breed mature female dogs were randomly selected and kept under the new situation for 3 weeks. All ovarian vessels of the dogs were double ligated, bilaterally. For 13 weeks, blood samples, vaginal cytology, and ultrasound examinations continued weekly. All dogs received eCG (200 IU/dog, IM, 5 days) and hCG (500 IU/case IM, 5th day) 1 month after ovarian ligation. After 4 weeks, all dogs were ovariohysterectomized, and ovarian specimens were submitted for histopathological evaluations. Estradiol 17β and progesterone were assayed with ELISA kits. Estradiol concentration decreased after ligation and continued to decrease to the end of study (during 10 weeks). Estradiol concentrations were different before ligation and after hormone stimulation and ovariohysterectomy (P < 0.01). Progesterone concentration decreased suddenly after ligation and had a plateau during the study. Progesterone concentrations were different before ligation period and after ligation and hormone stimulation periods (P < 0.03). An ultrasonographical evaluation showed normal ovarian structures before ligation and ovarian adhesions to bursa and ovarian hyperechogenicity without the presence of active structures. Bleeding, congestion, primordial and primary follicle necrosis, and some degree of fibrosis were observed in the histopathological evaluations. Bilateral whole vessel ovarian ligation could inactivate dogs’ ovarian function during 8 weeks. So, we suggest ligation of whole vessels of dogs’ ovaries as an alternative method for ovariectomy.

Production of Tetraploid Gynogenetic Loach Using Diploid Eggs of Natural Tetraploid Loach, Misgurnus anguillicaudatus, Fertilized with UV-Irradiated Sperm of Megalobrama amblycephala without Treatments for Chromosome Doubling

The gynogenesis phenomenon in nature mainly appears in the reproduction of fish and invertebrates. So far, gynogenesis has been successfully induced in many fish species with the aid of some physical or chemical methods for chromosome doubling. However, few fish can produce gynogenetic progenies, genetically identical or similar to the somatic cells of the mothers, without a treatment for the doubling of chromosomes, which may be related to apomixis, premeiotic endoreduplication, or premeiotic endomitosis. At present, no studies are available about fish with normal ovarian structures producing gynogenetic progenies that could spontaneously double their chromosomes. According to the analyses of flow cytometry, chromosome number, and microsatellites, we found that, with the use of UV-irradiated sperm of blunt snout bream Megalobrama amblycephala, tetraploid loach Misgurnus anguillicaudatus produced tetraploid gynogenetic progenies without any treatments for the doubling of chromosomes. To determine the genetic relationships of gynogenetic progenies and their maternal parent, microsatellite genotyping was conducted. The results indicated that the reason for spontaneous chromosome duplication in gynogenetic progenies may be cytokinesis or inhibition of the extrusion of the second polar body. This is the first report on fish with normal ovarian structures that can produce gynogenetic progenies which spontaneously double their chromosomes and which are genetically identical or similar to the somatic cells of the mothers.

Pcsk6 mutant mice exhibit progressive loss of ovarian function, altered gene expression, and formation of ovarian pathology

Bioactivation of precursor proteins by members of the proprotein convertase (PC) family is essential for normal reproduction. The Pcsk6 gene is a member of the PC family that is expressed in numerous ovarian cell types including granulosa cells and oocytes. We hypothesized that loss of PCSK6 would produce adverse effects in the mouse ovary. Mice incapable of expressing PCSK6 (Pcsk6(tm1Rob)) were obtained, and reproductive parameters (serum hormones, whelping interval, estrus cyclicity, and fertility) were compared to Pcsk6(+/+) mice. While Pcsk6(tm1Rob) female mice are fertile, they manifest reduced reproductive capacity at an accelerated rate relative to Pcsk6(+/+) mice. Reproductive senescence is typically reached by 9 months of age and is correlated with loss of estrus cyclicity, elevated serum FSH levels, and gross alterations in ovarian morphology. A wide range of ovarian morphologies were identified encompassing mild, such as an apparent reduction in follicle number, to moderate--ovarian atrophy with a complete absence of follicles--to severe, manifesting as normal ovarian structures replaced by benign ovarian tumors, including tubulostromal adenomas. Targeted gene expression profiling highlighted changes in RNA expression of molecules involved in processes such as steroidogenesis, gonadotropin signaling, transcriptional regulation, autocrine/paracrine signaling, cholesterol handling, and proprotein bioactivation. These results show that PCSK6 activity plays a role in maintaining normal cellular and tissue homeostasis in the ovary.

Genetic evidence against a role for W-linked histidine triad nucleotide binding protein (HINTW) in avian sex determination

Birds have a ZZ/ZW sex chromosome system, but the mechanism of sex determination remains unknown. The heterogametic sex is female (ZW) and one hypothesis holds that the W chromosome carries a dominant-acting ovary-determining gene. The strongest candidate ovary-determinant on the W chromosome is HINTW, which encodes an aberrant nucleotide hydrolase enzyme. HINTW is conserved amongst all carinate (flying) birds and it is strongly expressed in the gonads and other tissues of female chicken embryos. This and other lines of circumstantial evidence support the proposal that HINTW is the female-determining gene in birds. However, in vivo gain-of-function or loss-of-function studies have not hitherto been reported to test this hypothesis. We tested the potential role of HINTW by mis-expressing it in genetically male (ZZ) embryos, using the RCASBP avian retroviral vector. Strong, widespread expression was delivered throughout the embryo, including the urogenital system, as assessed by whole mount in situ hybridisation. This expression pattern mimicked that seen in normal ZW females, in which HINTW is widely expressed. Strong mis-expression was observed throughout the gonads of genetic male (ZZ) embryos. However, despite strong HINTW expression, ZZ gonads developed normally as bilateral testes. In tissue sections of ZZ urogenital systems transgenic for HINTW, normal testicular histology was observed. Female (ZW) gonads over-expressing HINTW also developed normally, with normal ovarian structure and left/right asymmetry. These results provide genetic evidence against a dominant role for HINTW in avian sex determination.

Ovarian Fibromatosis

Ovarian fibromatosis is a rare benign nonneoplastic condition with ovarian enlargement in young women and characterized by a proliferation of collagen-producing spindle cells surrounding normal ovarian structures. We reported magnetic resonance findings of a case that the affected ovarian parenchyma with follicles was surrounded by very low intense thick fibrous tissue on T2-weighted images such as "black garland" around the ovary. The magnetic resonance findings well reflected the pathological feature of the disease and may be diagnostic.

Transplantation of frozen–thawed late-gestational-age human fetal ovaries into immunodeficient mice

To compare the development of human fetal follicles from late-gestational-age fetuses frozen-thawed gradually and slowly with dimethylsulfoxide (DMSO) or 1,2 propanediol (PROH) and sucrose after renal grafting into follicle stimulating hormone-treated immunodeficient mice. Controlled histologic study of grafted human fetal ovaries. Major tertiary care academic center. Eleven women undergoing pregnancy termination at 22 to 33 gestational weeks. None. Microscopic morphometric analysis and immunohistochemistry for proliferating cell nuclear antigen (PCNA). Only follicles from samples frozen-thawed with PROH developed to secondary and antral stages 4 to 6 months after grafting, with PCNA expression in their granulosa cells. However, the number of surviving/developing follicles per section was very low (4-25 per graft), compared with 71 to 406 follicles in pretransplantation samples. Graft recovery was very high, with similar rates for transplants frozen-thawed with PROH and DMSO. Normal ovarian structure after grafting was identified only in the PROH frozen-thawed grafts. In deteriorated grafts, frozen-thawed with either DMSO or PROH, net-like hollows replaced follicles, whereas tubule-like structures were only identified in DMSO frozen-thawed grafts. This is the first report of the development of late-pregnancy-stage human fetal follicles in immunodeficient mice. PROH freezing-thawing supported development and survival better than DMSO. However, the low follicular survival points to the urgent need for efficient methods to enhance vascularization rate and prevent ischemia.

Estrogen Receptors α and β and Progesterone Receptors in Normal Bovine Ovarian Follicles and Cystic Ovarian Disease

The purpose of this study was to determine by immunohistochemistry the expression of estrogen and progesterone receptors in ovarian follicular structures from cows with cystic ovarian disease (COD) and to compare these with normal ovarian structures. Secondary, tertiary, atretic, and cystic follicles were evaluated. The follicular cysts of animals with COD presented a significantly higher expression of estrogen receptor alpha in all follicular layers than secondary, tertiary, and atretic follicles in both groups (P < .05). The intensity of estrogen receptor beta in the granulosa cell layer was stronger in tertiary than in secondary and atretic follicles in normal animals (P < .05) and in growing and cystic follicles in animals with COD (P < .05). Theca cells were scarcely stained in the 2 groups. Growing follicles and cysts from COD animals were less stained than tertiary follicles from normal animals (P < .05). Differences did not exist between the 2 groups with regard to the progesterone receptor. Ovaries of animals with COD exhibited altered estrogen receptors expression compared with that in normal animals.

Normal Ovarian Structure and Function with Normal Glucose Tolerance in Girls with Early Treatment of Classic Congenital Adrenal Hyperplasia

Hyperandrogenism and hyperinsulinism have both been suggested as etiologic factors behind functional ovarian hyperandrogenism or polycystic ovary syndrome. Females with congenital adrenal hyperplasia provide a clinical model to evaluate the contribution of pre- and post-natal hyperandrogenism on ovarian structure and function. Study Objective To evaluate glucose tolerance, and structure and androgen production of the ovaries in young females with classic congenital adrenal hyperplasia. Design, Setting, Participants Cross-sectional study, including the enrollment of participants, ages 8 to 20 years, recruited from the pediatric endocrinology clinical program of a tertiary pediatric referral center. Interventions Ten participants had oral glucose tolerance testing, adrenal and ovarian androgen measurements, and pelvic ultrasound studies performed. Main Outcome Measures Presence of altered response to glucose challenge, ovarian hyperandrogenism, or presence of polycystic ovaries by ultrasound. Results Measurements of fasting blood glucose, post-challenge glucose, and insulin resistance were normal in this sample. There was no evidence of ovarian hyperandrogenism after adrenal suppression with dexamethasone. All participants had normal ovarian structure without evidence of polycystic ovaries. Conclusions Females with classic congenital adrenal hyperplasia (21-hydroxylase deficiency) and normal glucose tolerance appear to have normal ovarian structure and function during the peripubertal period.

Fluorescence spectroscopy for in vivo characterization of ovarian tissue.

The objective of this study was to explore whether fluorescence spectroscopy signatures differed between normal variations within the ovary, benign neoplasms, and ovarian cancer. Ovarian tissue fluorescence emission spectra were collected sequentially at 18 excitation wavelengths ranging from 330 to 500 nm from 11 patients undergoing oophorectomy and assembled into fluorescence excitation emission matrices (EEMs); biopsies corresponded to the area interrogated. Spectral areas that could differentiate normal ovary, benign neoplasms, and cancers were evaluated, using histopathology as the reference standard. The most promising measurements are (1) the integrated fluorescence intensity from 400 to 430 nm excitation at 460 nm emission, and (2) the ratios of fluorescence intensities at 330 nm excitation, 385 and 500 nm emission, and at 375 and 415 nm excitation, 460 nm emission. Simple systems to visualize these optical signatures at laparoscopy could be designed. Fluorescence spectroscopy may have the ability to distinguish ovarian cancers from normal ovarian structures and benign neoplasms, as well as differentiate between normal variations and metaplastic structures and should be further explored as a device for the early detection of ovarian cancers.

Suppressive effect of fetal testes on development of fetal ovaries transplanted into adult males in the rat.

The age-related testicular effect on the ovarian primordia was studied by combined transplantation of fetal testes and ovaries in adult male hosts. First, ovarian primordia of 14-day fetal rats were transplanted into a renal subcapsular position of castrated or intact adult male rats. In both the castrated and the intact hosts, most of the ovarian transplants developed normally with only 3 of them having in part seminiferous tubule-like structures in addition to normal ovarian structure. Second, a 14-day ovary was combined with a fetal testis the age of which varied from 13- to 18-day, and the combination was transplanted. In the combination of a 14-day ovary and a 13-day testis, the results varied in such a way that the ovary or the testis alone developed or otherwise, both gonads developed well. In union with 15- to 18-day testes, the ovaries did not develop, although the testes developed well. These results suggest that the 14-day ovarian primordia have a slight reactiveness to androgens of host rats and that the 13-day fetal testes begin to inhibit the development of the 14-day ovaries co-transplanted with them.

Ultrasound as an aid to diagnosis of granulosa cell tumour in the mare

The ultrasonic appearance of seven cases of granulosa cell tumour is described, and compared with an ovarian haematoma and an ovarian serous cystadenoma. The granulosa cell tumours varied from being uniformly dense to having one or several large fluid filled cysts. Some tumours resembled the haematoma or cystadenoma and also, in some aspects, normal ovarian structures such as corpora haemorrhagica or follicles in early pregnancy. There was no typical ultrasonographic appearance of the granulosa cell tumours which enabled definitive diagnosis. However, ultrasound may be a diagnostic aid when used in conjunction with the history and findings on rectal palpation.