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PubMed1984

Lectin binding sites of the mouse ovary, intraovarian and ovulated ova.

Fluorescein isothiocyanate (FITC) labeled lectins were used to study the distribution of specific binding sites in histologic sections of mouse ovaries as well as ovulated ova. Four distinct patterns of reactivity of the components of the follicle (exclusive of the ovum) and the surrounding ovarian stroma were recognized: uniform staining of granulosa cells, theca cells and surrounding stroma; weak to moderate staining of the granulosa cells and strong staining of the theca cells and stromal cells; no reactivity of the granulosa cells coupled with strong reactivity of the theca and stromal cells; no reactivity with any component of the cumulus oophorus. Three lectins (from Triticum vulgare, Arachis hypogaea and Maclura pomifera) distinctly accentuated the basal lamina of the cumulus oophorus. The reaction of lectins with oocytes and zona pellucida revealed six distinct patterns: no reactivity with either structure; weak reactivity with the cytoplasm of the oocyte and no reactivity with the zona pellucida; very strong reactivity with the cytoplasm of the oocyte and no reactivity with the zona pellucida; moderate reactivity with both the oocyte and the zona pellucida; moderate reactivity with the oocyte and very strong reactivity with the zona pellucida; no reactivity with the oocyte and moderate reactivity with the zona pellucida. The same pattern of reactivity was seen in the ovulated ova in the oviduct except for the lectin from Arachis hypogaea, the reactivity of which changed upon ovulation and/or fertilization of the ovum. These data provide a list of lectin markers for distinct components of the mouse ovary.

Animals↗
PubMed1978

Maturational changes in sheep ovarian follicles: gonadotrophic stimulation of cyclic AMP production by isolated theca and granulosa cells.

Theca and granulosa tissues isolated from sheep ovarian follicles of different sizes were incubated in the presence of human chorionic gonadotrophin (HCG; 5 IU/ml) or follicle stimulating hormone (FSH; 5 microgram NIH-FSH-S11/ml) for 40 min. Changes in the total amounts of cyclic 3',5'-adenosine monophosphate (cAMP) were used as an index of the responsiveness of these preparations to the hormones. Thecal tissue of both large (4-6 mm in diameter) and small (1-3 mm) follicles responded similarly to gonadotrophins. Granulosa cells from small follicles failed to respond to stimulation by HCG. FSH, however, consistently increased cAMP production in comparison with controls or cells treated with HCG. Granulosa cells of large follicles responded to both HCG and FSH.

Animals↗
PubMed1994

Regulation of steroid production in ovarian stromal tissue from 5- to 8-week-old pullets and laying hens.

Studies were conducted to evaluate the regulation of steroid production in dispersed cells from ovarian stromal tissue from 5- to 8-week-old-pullets (IM cells) and laying hens (MAT cells). Short-term incubation of IM and MAT cells with ovine (o) LH resulted in a dose-dependent increase in progesterone, androstenedione and oestradiol production; progesterone production was greater in MAT cells than in IM cells (P < 0.05) in response to 2-200 ng oLH ml-1, whereas androstenedione and oestradiol production was greater in MAT cells following treatment with 20 and 200 ng oLH ml-1 (P < 0.05). In both cell populations the cyclic adenosine monophosphate (cAMP) analogue, 8-bromo-cAMP (1 and 10 mmol l-1) stimulated progesterone and androstenedione production, whereas oLH (200 ng ml-1) and forskolin (1-10 mumol l-1) promoted cAMP accumulation (P < 0.05 compared with basal values). However, treatment with the protein kinase C activator, phorbol 12-myristate 13-acetate (PMA), did not alter basal or oLH-stimulated cAMP accumulation or progesterone production in either IM or MAT cells (P > 0.10). PMA did, however, inhibit agonist-induced androstenedione production (P < 0.05); co-treatment with the calcium ionophore A23187 potentiated this inhibitory effect. Finally, treatment with transforming growth factor-alpha (TGF-alpha; 1.8-18 pmol l-1) did not affect basal or oLH-stimulated progesterone or androstenedione production by IM cells, MAT cells, theca cells from 6-8 mm follicles or theca cells from the second largest (F2) follicle (P > 0.10).(ABSTRACT TRUNCATED AT 250 WORDS)

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed2004

Minireview: transcriptional regulation of gonadal development and differentiation.

The embryonic gonad is undifferentiated in males and females until a critical stage when the sex chromosomes dictate its development as a testis or ovary. This binary developmental process provides a unique opportunity to delineate the molecular pathways that lead to distinctly different tissues. The testis comprises three main cell types: Sertoli cells, Leydig cells, and germ cells. The Sertoli cells and germ cells reside in seminiferous tubules where spermatogenesis occurs. The Leydig cells populate the interstitial compartment and produce testosterone. The ovary also comprises three main cell types: granulosa cells, theca cells, and oocytes. The oocytes are surrounded by granulosa and theca cells in follicles that grow and differentiate during characteristic reproductive cycles. In this review, we summarize the molecular pathways that regulate the distinct differentiation of these cell types in the developing testis and ovary. In particular, we focus on the transcription factors that initiate these cascades. Although most of the early insights into the sex determination pathway were based on human mutations, targeted mutagenesis in mouse models has revealed key roles for genes not anticipated to regulate gonadal development. Defining these molecular pathways provides the foundation for understanding this critical developmental event and provides new insight into the causes of gonadal dysgenesis.

Animals↗
PubMed2005

Expression of Smad4 during rat ovarian development.

OBJECTIVE: To clarify the signal transduction pathway of transforming growth factor-betasuperfamily (TGF-betas) in the regulation of follicle growth by investigating the expressions of Smad4 protein and mRNA in rat ovaries in different developmental stages. METHODS: Rat ovaries of different developmental stages were obtained to determine the expression of Smad4 protein by immunohistochemistry and image analysis system, with Smad4 mRNA measured by semi-quantitative RT-PCR. Specific primers of Smad4 and GAPDH (internal control) were used for amplification by RT-PCR, and the ratios of their integrated optical densities were calculated to estimate the relative quantity of Smad4 mRNA expression. RESULTS: Smad4 protein was widely expressed in the ovary, mainly in the follicles, and the location and intensity of Smad4 expression varied with the degree of maturation of the ovary. In the early developmental stages, Smad4 protein expressed mainly in the primordial and preantral follicles, but little in the stromal cells, and its expression intensity in the stroma increased gradually in the course of ovarian maturation. After sexual maturity, Smad4 expression intensity varied only insignificantly among the granulosa cells, theca cells and stromal cells of the antral and mature follicles (P>0.05). The staining intensity of Smad4 in the follicles also underwent changes in relation with their development, being less intense in the oocytes of the antral and matured follicles as compared to the preantral follicles (P<0.05 and P<0.01, respectively) but markedly greater in the theca cells of the antral and matured follicles than in the preantral follicles (P<0.01). No significant difference in Smad4 expression was found in the granulosa cells of different developmental stages (P>0.05). RT-PCR demonstrated that Smad4 mRNA was expressed in all the developmental stages of the rat ovary; and from the 3rd week on, the integrated optical density of Smad4 and GAPDH was significantly higher than that in 1-day-old neonatal rats. CONCLUSION: The expression patterns of Smad4 protein and mRNA in rat ovary in the course of its development indicate that Smad signal transduction may play a role in the folliculogenesis.

Animals↗
PubMed2002

Enhanced serum oestrogen levels and highly steroidogenic, luteinized atretic follicles in the ovaries of the Djungarian hamster (Phodopus sungorus) kept under a short photoperiod from birth.

OBJECTIVE: In contrast to the elaborate information available on the effects of the photoperiod on the testes of hamsters, little is known about the influence on their ovaries. This study aimed to describe the ovarian follicular development and steroid hormone production in Djungarian hamsters kept from birth under a short daylight regime. DESIGN AND METHODS: Female Djungarian hamsters (Phodopus sungorus) were kept under two different light regimes: (i) 16 h light:8 h darkness (long daylight; LD) and (ii) 4 h light:20 h darkness (short daylight; SD). They were killed at 28, 56 and 80 days after birth; blood and ovaries were collected. Ovaries were either fixed in Bouin's solution or frozen. Fixed material was dehydrated, embedded in paraffin, serially sectioned at 5 microm and stained with haematoxylin and eosin, whereafter all healthy and atretic follicles were classified and counted. 3beta-Hydroxysteroid dehydrogenase (3beta-HSD) was histochemically demonstrated in 10 microm sections of frozen ovaries. Serum oestradiol-17beta and progesterone levels were determined by RIA. RESULTS: The numbers of healthy preantral and antral follicles were higher in LD than in SD hamsters. Antral follicles did not significantly differ in number during development in LD hamsters, but they were completely absent from 80-day-old SD animals. In LD animals the number of apoptotic preantral follicles dramatically increased with age. In SD animals the numbers of apoptotic antral follicles strongly decreased with age, whereas numerous non-apoptotic follicles with luteinized granulosa cells and a degenerated oocyte appeared, and in increasing numbers with age. During development, moderate 3beta-HSD activity was present in interstitial cells, theca cells of healthy follicles, and in both theca and granulosa cells of degenerating follicles. Strong enzyme activity was found in the hypertrophied granulosa cells of luteinized atretic follicles. Mean serum progesterone values varied from 2 to 6 nmol/l and were not different in LD and SD hamsters. Mean serum oestradiol levels varied from 132 to 542 and 325 to 2353 pmol/l in LD and SD hamsters respectively. The highest oestradiol levels were found in SD animals at day 28 of development. CONCLUSIONS: Folliculogenesis was dramatically disturbed in Djungarian hamsters raised under a short photoperiod. These animals developed high serum oestradiol levels and numerous luteinized atretic follicles with highly steroidogenic granulosa cells, which appear to be the source of the increased serum oestradiol levels.

3-Hydroxysteroid Dehydrogenases↗
PubMed1995

Sclerosing stromal tumour in a cow.

A case of sclerosing stromal tumour in an 8-year-old cow is described. The left ovary was entirely replaced by neoplastic tissue, whose features were pseudolobulation of cellular areas, intercellular oedema and calcified plaques. The neoplastic cells varied in morphology and were similar to ovarian stromal cells, theca cells or lutein cells. Immunohistochemically, S-100 protein, oestradiol and testosterone were detected in theca-like or luteinized cells, some of which had ultrastructural features suggestive of steroid hormone synthesis. In contrast, desmin and actin characterized slender spindle cells. This is the first report of a sclerosing stromal tumour in cattle.

Animals↗
PubMed2005

LH/hCG-stimulated androgen production and selective HDL-cholesterol transport are inhibited by a dominant-negative CREB construct in primary cultures of rat theca-interstitial cells.

Theca-interstitial (T-I) cells synthesize androgens that are converted to estrogen by the granulosa cells. In rat ovary, T-I cells primarily utilize HDL-derived cholesteryl esters (CE) as a precursor for androgen synthesis. The HDL-CE is delivered to steroidogenic cells by a process termed "selective" uptake in which CE is internalized without the simultaneous uptake of apolipoprotein(s). This process is mediated by an HDL receptor, scavenger receptor class B, type I (SR-BI) and is stimulated by trophic hormone (LH/hCG), which also activates the cAMP cascade. In this study, we tested whether the adenoviral (Ad)-mediated introduction of a dominant-negative analog of cyclic AMP response element binding protein (A-CREB) inhibits the stimulatory effect of LH/hCG on the selective uptake of high-density lipoprotein (HDL)-cholesterol and androgen production in primary cultures of rat T-I cells. Androstenedione production by cultured T-I cells was stimulated by hCG and by the adenoviral overexpression of wtCREB. Additionally, the stimulatory effect observed with hCG was amplified in the presence of HDL. Androgen synthesis was increased 17-fold in the presence of HDL and hCG but the stimulatory effect of hCG was inhibited by Ad A-CREB by approx 70%. In the selective up-take studies, cell-surface association of the labeled HDL was significantly enhanced by hCG treatment, and this effect was inhibited by Ad A-CREB. The selective uptake of HDL-cholesterol was also enhanced by hCG but exposure to Ad A-CREB also abrogated this effect. It is concluded that CREB plays an intermediary role in the stimulatory action of LH/hCG on androgen synthesis and the selective uptake of HDL-cholesterol in T-I cells.

Adenoviridae↗
PubMed1976

Enzyme histochemistry of cultured ovarian cells. II. Histochemistry of porcine theca interna cells in tissue culture.

By means of histochemical technique the activities of delta53beta-, 17beta-, 20alpha-hydroxysteroid dehydrogenases and glucose-6-phosphate dehydrogenase, as well as alkaline and acid phosphatase were investigated in monolayer cultures of theca interna cells, isolated from preovulatory porcine ovarian follicles. It was found that theca interna cells exhibited high and constant activity of delta53beta-hydroxysteroid dehydrogenase and G6P-DH, whereas activities of both 17beta- and 20alpha-hydroxysteroid dehydrogenase were lower and showed some fluctuations during in vitro culture. Addition of LH to the medium brought about the increase of all studied dehydrogenases. FSH was less effective. Estradiol showed quite and inhibiting effect. All the hormones mentioned above caused the increase of alkaline and acid phosphatase activity in cultured porcine theca interna cells.

17-Hydroxysteroid Dehydrogenases↗
PubMed1990

Immunohistochemical localization of growth hormone-releasing hormone in human gonads.

Human gonads were examined for the presence of ir growth hormone-releasing-hormone. We demonstrated the presence of immunostainable cells both in ovarian and in testicular tissue using a non-crossreactive anti-GRH antiserum and the immunoperoxidase detection technique. In ovaries from ovulating women, GRH immunoreactivity was localized in the corpora lutea; granulosa cells, theca cells and cells of primary follicles did not stain. In premenopausal ovaries, staining was detectable in scattered luteinized stromal cells. In testes from post-puberal men GRH immunoreactivity was localized in the Leydig cells; cells of the germinal epithelium did not stain. These results demonstrate the presence of GRH in human gonads and suggest that this peptide may exert regulatory function at the testicular and ovarian levels.

Female↗
PubMed1984

Malignant sex cord-stromal tumor of testis.

We report a case of sex cord-stromal tumor in the testis, which metastasized to the retroperitoneal space, resulting in death 13 months after orchiectomy. The primary lesion consisted of granulosa cells, theca cells, Sertoli cells and undifferentiated gonadal stroma. Biopsy of the retroperitoneal metastasis showed predominance of the Sertoli cell component.

Adult↗
PubMed2001

Stimulatory and synergistic effects of luteinising hormone and insulin like growth factor 1 on the secretion of vascular endothelial growth factor and progesterone of cultured bovine granulosa cells.

Vascular endothelial growth factor (VEGF) is the most important factor in the regulation of angiogenesis. Associated with luteinisation and formation of corpus luteum (CL) are alterations in luteal vascularity. The aim of the study was to test under in vitro conditions the stimulation of VEGF and progesterone (P) secretion of bovine granulosa cells by LH, IGF1 (insulin like growth factor) or by factors known to be produced by luteinised granulosa cells or in the early CL. Localisation of VEGF protein in preovulatory follicle and early CL were achieved by immunohistochemistry. LH and IGF1 stimulated dose dependently and significantly P and VEGF when tested alone. Both hormones added simultaneously had clear additive and even more interesting far greater (synergistic) effects on P with LH (0.1 ng/ml) plus 5 or 10 ng IGF1. In contrast, VEGF was stimulated only additively with 0.1 ng/ml of LH plus 5 or 10 ng IGF1. But with the higher dose of LH (1 ng/ml) additionally to the additive effect a tendency for a synergistic action (which was significant with 1 ng LH plus 5 ng IGF1/ml) was observed. Endothelin, oxytocin, progesterone, atrial natiuretic peptide, angiotensin II, prostaglandin F2 alpha alpha, prostaglandin E2, cortisol, fibroblast growth factor 1 and 2 and growth hormone showed no effect neither on P nor on VEGF. Tumour necrosis factor alpha (TNF alpha) stimulated (P < 0.05) VEGF with 10 or 100 ng/ml but not P. TPA (12-0 tetra decaenoyl-phorbol-13-acetate) or Ca2+ ionophore did not show a stimulatory effect in contrast to forskolin which increased P and VEGF secretion dose dependently. The VEGF protein was localised in follicle (granulosa cells, theca cells and some endothelial cells) and early (about 24 h after ovulation) CL (granulosa-lutein cells and endothelial cells). The same signalling pathway by stimulation of cAMP production and proteinkinase A activation for luteinisation and neo-vascularisation demonstrates a close temporal and spatial relationship of these normal physiological processes.

Animals↗