LncRNA-m18as1 competitively binds with miR-18a-5p to regulate follicle-stimulating hormone secretion through the Smad2/3 pathway in rat primary pituitary cells / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Long noncoding RNAs (lncRNAs) are expressed in different species and different tissues, and perform different functions, but little is known about their involvement in the synthesis or secretion of follicle-stimulating hormone (FSH). In general, we have revealed lncRNA‍‒‍microRNA (miRNA)‍‒‍‍messenger RNA (mRNA) interactions that may play important roles in rat primary pituitary cells. In this study, a new lncRNA was identified for the first time. First, we analyzed the gene expression of lncRNA-m18as1 in different tissues and different stages by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and observed the localization of lncRNA-m18as1 with fluorescence in situ hybridization, which indicated that this lncRNA was distributed mainly in the cytoplasm. Next, we used RT-qPCR and enzyme-linked immunosorbent assay (ELISA) to analyze the regulation of FSH synthesis and secretion after overexpression or knockdown of lncRNA-m18as1 and found that lncRNA-m18as1 was positively correlated with FSH synthesis and secretion. In addition, mothers against decapentaplegic homolog 2 (Smad2) was highly expressed in our sequencing results. We also screened miR-18a-5p from our sequencing results as a miRNA that may bind to lncRNA-m18as1 and Smad2. We used RNA immunoprecipitation-qPCR (RIP-qPCR) and/or dual luciferase assays to confirm that lncRNA-m18as1 interacted with miR-18a-5p and miR-18a-5p interacted with Smad2. Fluorescence in situ hybridization (FISH) showed that lncRNA-m18as1 and miR-18a-5p were localized mainly in the cytoplasm. Finally, we determined the relationship among lncRNA-m18as1, miR-18a-5p, and the Smad2/3 pathway. Overall, we found that lncRNA-m18as1 acts as a molecular sponge of miR-18a-5p to regulate the synthesis and secretion of FSH through the Smad2/3 pathway.

Altered microRNA profiles of testicular biopsies from patients with nonobstructive azoospermia / 亚洲男科学杂志(英文版)

Many studies have shown that microRNAs (miRNAs) play vital roles during the spermatogenesis. However, little is known about the altered miRNA profiles of testicular tissues in nonobstructive azoospermia (NOA). Using microarray technology, the miRNA expression profiles of testicular biopsies from patients with NOA and of normal testicular tissues were determined. Bioinformatics analyses were conducted to predict the enriched biological processes and functions of identified miRNAs. The microarray data were validated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), the results of which were then validated with a larger sample size. Correlations between the miRNA expression levels and clinical characteristics were analyzed. Receiver operating characteristic (ROC) curve analysis was used to evaluate the diagnostic ability of miRNAs for azoospermia. Hierarchical clustering showed that 129 miRNAs were significantly differentially expressed between the NOA and control groups. Bioinformatics analysis indicated that the differentially expressed miRNAs were involved in spermatogenesis, cell cycle, and mitotic prometaphase. In the subsequent qRT-PCR assays, the selected miRNA expression levels were consistent with the microarray results, and similar validated results were obtained with a larger sample size. Some clinical characteristics were significantly associated with the expression of certain miRNAs. In particular, we identified a combination of two miRNAs (miR-10b-3p and miR-34b-5p) that could serve as a predictive biomarker of azoospermia. This study provides altered miRNA profiles of testicular biopsies from NOA patients and examines the roles of miRNAs in spermatogenesis. These profiles may be useful for predicting and diagnosing the presence of testicular sperm in individuals with azoospermia.

FSH receptor binding inhibitor up-regulates ARID1A and PTEN genes associated with ovarian cancers in mice

Experiments were conducted to determine if the follicle-stimulating hormone (FSH) receptor binding inhibitor (FRBI) impacts the expression levels of AT-rich interactive domain-containing protein 1A (ARID1A) and phosphatase and tensin homolog (PTEN) in ovaries and blood, as well as expressions of follicle-stimulating hormone cognate receptor (FSHR) gene and proteins. Mice in FRBI-10, FRBI-20, FRBI-30, and FRBI-40 groups were intramuscularly injected with 10, 20, 30, and 40 mg FRBI/kg, respectively, for five consecutive days. Western blotting and qRT-PCR were utilized to determine expression levels of ARID1A and PTEN proteins and mRNAs. Serum ARID1A and PTEN concentrations of the FRBI-40 group were higher than the control group (CG) and FSH group (P<0.05). FSHR mRNA levels of FRBI-20, FRBI-30, and FRBI-40 groups were lower than that of CG and FSH groups on day 15 (P<0.05 or P<0.01). Expression levels of FSHR proteins of FRBI-30 and FRBI-40 groups were lower than those of CG and FSH groups (P<0.05). Levels of ARID1A and PTEN proteins of the FRBI-30 group were greater than CG on days 20 and 30 (P<0.05). FRBI doses had significant positive correlations to levels of ARID1A and PTEN proteins. Additionally, ARID1A and PTEN had negative correlations to FSHR mRNAs and proteins. A high dose of FRBI could promote the expression levels of ARID1A and PTEN proteins in ovarian tissues. FRBI increased serum concentrations of ARID1A and PTEN. However, FRBI depressed expression levels of FSHR mRNAs and proteins in mouse ovaries.

Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles

The objective this study was to determine the effect of phytohemagglutinin (PHA) on survival, growth and gene expression in caprine secondary follicles cultured in vitro. Secondary follicles (∼0.2 mm) were isolated from the cortex of caprine ovaries and cultured individually for 6 days in α-MEM+ supplemented with PHA (0, 1, 10, 50, 100, or 200 µg/mL). After 6 days of culture, follicle diameter and survival, antrum formation, ultrastructure and expression of mRNA for FSH receptors (FSH-R), proliferating cell nuclear antigen (PCNA), and neuronal nitric oxide synthase were determined. All treatments maintained follicular survival [α-MEM+ (94.59%); 1 µg/mL PHA (96.43%); 10 µg/mL PHA (84.85%); 50 µg/mL PHA (85.29%); 100 µg/mL PHA (88.57%), and 200 µg/mL PHA (87.50)], but the presence of 10 µg/mL PHA in the culture medium increased the antrum formation rate (21.21%) when compared with control (5.41%, P < 0.05) and ensured the maintenance of oocyte and granulosa cell ultrastructures after 6 days of culture. The expression of mRNA for FSH-R (2.7 ± 0.1) and PCNA (4.4 ± 0.2) was also significantly increased in follicles cultured with 10 µg/mL PHA in relation to those cultured in α-MEM+ (1.0 ± 0.1). In conclusion, supplementation of culture medium with 10 µg/mL PHA maintains the follicular viability and ultrastructure, and promotes the formation of antral cavity after 6 days of culture in vitro.

Influence of nitric oxide on in vitro growth, survival, steroidogenesis, and apoptosis of follicle stimulating hormone stimulated buffalo (Bubalus bubalis) preantral follicles

Effect of sodium nitroprusside (SNP), a nitric oxide (NO) donor, on in vitro survival, growth, steroidogenesis, and apoptosis of buffalo preantral follicles (PFs) was investigated. PFs (200~250 microm) were isolated by micro-dissection and cultured in 0 (control), 10(-3), 10(-5), 10(-7), and 10(-9) M SNP. To examine the reversible effect of SNP, PFs were cultured with 10(-5) M SNP + 1 mM Nomega-nitro-L-arginine methyl ester (L-NAME) or 1.0 microg hemoglobin (Hb). The results showed that greater concentrations of SNP (10(-3), 10(-5), 10(-7) M) inhibited (p < 0.05) FSH-induced survival, growth, antrum formation, estradiol production, and oocyte apoptosis in a dose-dependent manner. However, a lower dose of SNP (10(-9) M) significantly stimulated (p < 0.05) the survival, growth, antrum formation, follicular oocyte maturation, and stimulated progesterone secretion compared to the control. A combination of SNP + L-NAME promoted the inhibitor effect of SNP while a SNP + Hb combination reversed this effect. Nitrate and nitrite concentrations in the culture medium increased (p < 0.05) in a dose-dependent manner according to SNP concentration in the culture medium. At higher concentrations, SNP had a cytotoxic effect leading to follicular oocyte apoptosis whereas lower concentrations have stimulatory effects. In conclusion, NO exerts a dual effect on its development of buffalo PFs depending on the concentration in the culture medium.

Influência da proteína de soja enriquecida com isoflavona e dos exercícios com pesos sobre o metabolismo energético de mulheres na pós-menopausa / Influence of soy protein and exercises with weights on the energy metabolism of women in post-menopause

O objetivo do estudo foi analisar a influência da ingestão alimentar de proteína da soja enriquecida com isoflavona e dos exercícios com pesos sobre o metabolismo energético de mulheres na pós-menopausa. Casuística e métodos. Ensaio clínico, 16 semanas, envolvendo 60 mulheres, 59 (7) anos, distribuídas em 4 grupos: G1 (proteína da soja e exercício), G2 (placebo e exercício), G3 (proteína da soja e sem exercício) e G4 (placebo e sem exercício). A proteína da soja e o placebo (maltodextrina) foram distribuídos, aleatoriamente, sob a forma de pó, na porção de 25 gramas/dia, enriquecida com 50 mg de isoflavona. Foram 10 exercícios com pesos, realizados em 3 sessões semanais, com 3 séries de 8-12 repetições cada, carga de 60-80% de 1 repetição máxima (RM). O metabolismo energético foi analisado a partir do gasto energético de repouso (GER) e do quociente respiratório (QR). O GER foi calculado utilizando o O2 e o CO2, obtidos por calorimetria indireta (Quinton-QMC®), durante 30 minutos, sob temperatura e umidade controladas, e o QR pela relação CO2/O2. Foram utilizadas variáveis de controle: idade, hormônio folículo estimulantes (FSH), proteína e valor calórico total da alimentação (VCT), massa muscular e índice de massa corporal IMC (kg/m2). Na análise estatística foram utilizados: ANOVA, teste "t" de Student e regressão múltipla, por meio do software Stata 9.2, α<0,05. Resultados. As mulheres apresentaram homogeneidade em todas as variáveis do estudo. Houve aumento, significante (p<0,05) do GER no G1 (158 kcal/dia) e G2 (110 kcal/dia), correspondente a 17% e 9%, respectivamente, sem alteração do QR. Não houve associação entre as variáveis de controle, com o GER e o QR, entretanto, com a intervenção, observou-se aumento significante (p<0,05) da massa muscular, G1 (7%) e G2 (10%), e da ingestão proteica, G1 (de 19 para 25%) e G3 (de 19 para 26%), sem diferença entre grupos, quanto ao VCT. Conclusão. Exercícios com pesos são determinantes para o aumento do GER de mulheres na pós-menopausa, podendo ser potencializado pela ingestão de proteína da soja enriquecida com isoflavona

The objective of this study was analyze the influence of dietary ingestion of soy protein enriched with isoflavones and exercises with weights on energy metabolism of women in post-menopause. Material and methods. Clinical trial, 16 weeks, involving 60 women, 59 (7) years, divided into four groups: G1 (soy protein and exercise), G2 (placebo and exercise), G3 (without soy protein and exercise) and G4 (placebo and without exercise). Randomly, the soy protein and placebo (maltodextrin) were distributed in powder form; the portion of 25 grams / day was enriched with 50 mg of isoflavones. There were 10 exercises with weight, performed in three sessions per week with 3 sets of 8-12 repetitions each, with load of 60-80% of one maximum repetition (RM). The energy of metabolism was analyzed from the resting energy expenditure (REE) and respiratory quotient (RQ). The REE was calculated using the O2 and CO2, obtained through indirect calorimetry (QMC, Quinton ®) for 30 minutes under controlled temperature and humidity, and the ratio QR CO2/O2. Some variables to control the REE were used: age, follicle stimulating hormone (FSH), protein and total caloric content of food (VCT), muscle mass and body mass index - BMI (kg/m2). To the statistical analysis were used: ANOVA, "t" test of Student and multiple regression using the software Stata 9.2, α<0.05. Results. The women presented homogeneity for all variables of the study. Significantly, there was an increase (p <0.05) of SGA in G1 (158 kcal / day) and G2 (110 kcal/ day), corresponding to 17% and 9% respectively, and no alteration of QR. Between the variables control, REE and RQ, wasnt found an association, however, through the intervention, was observed a significant (p <0.05) muscle mass, G1 (7%) and G2 (10%), and ingestion of protein, G1 (from 19 to 25%) and G3 (from 19 to 26%), with no difference among groups toward the VCT. Conclusion. The exercises with weights are crucial to the increase of REE in women in post- menopause, and can be potentiated by the ingestion of soy protein enriched with isoflavones

Estradiol transdérmico e perfil lipídico: efeitos em um grupo específico de mulheres brasileiras pós-menopausadas / Transdermal estradiol and lipid profile: effects on a specific group of brazilian postmenopausal women / Estradiol transdérmico y perfil lipídico: efectos en un grupo específico de mujeres brasileñas posmenopáusicas

Fundamento: Em mulheres pós-menopausadas, mudanças significantes ocorrem, que podem induzir doenças cardiovasculares, tais como o perfil lipídico aterogênico devido a um aumento nos níveis de colesterol total e LDL, e uma diminuição nos níveis de HDL. A terapia de reposição hormonal (TRH) pode evitar essas mudanças no perfil lipídico. Objetivo: Determinar os efeitos da TRH constituída por estradiol transdérmico e acetato de medroxiprogesterona nos parâmetros bioquímicos e lipídicos de mulheres brasileiras pós-menopausadas. Métodos: Este é um estudo prospectivo, longitudinal, aberto, no qual trinta mulheres pós-menopausadas receberam estradiol em gel transdérmico (1 mg/dia) de forma contínua, combinado com acetato de medroxiprogesterona (MPA) (5 mg/dia) por 12 dias/mês. Os seguintes parâmetros foram determinados: colesterol total, triglicérides, lipoproteína de alta densidade (HDL-colesterol), lipoproteína de baixa densidade (LDL-colesterol), lipoproteína de muito baixa densidade (VLDL-colesterol), glicose, aspartato aminotransferase (AST), alanina aminotransferase (ALT), gama-glutamil transferase (GGT) e hormônio folículo estimulante (FSH). Resultados: Os parâmetros do perfil lipídico mostraram uma diminuição não-significante, enquanto os níveis de GGT e FSH apresentaram uma diminuição estatisticamente significante. Conclusões: O tratamento com estradiol em gel transdérmico não mostrou um impacto significante no perfil lipídico, de forma que não resultou em um efeito benéfico nos marcadores de doenças cardiovasculares, sugerindo que a dose, modo de administração e o tempo de tratamento foram importantes para esses resultados. Além disso, o tratamento com dose baixa e modo de administração transdérmico também demonstrou um significante efeito hepático nessa população. Dessa forma, esse tratamento pode fornecer efeitos interessantes sobre o perfil lipídico em mulheres brasileiras pós-menopausadas.

Background: In postmenopausal women, significant changes occur that can induce cardiovascular diseases, such as atherogenic lipid profile, due to an increase in total cholesterol and LDL levels, and a decrease in HDL cholesterol levels. The hormone replacement therapy (HRT) can prevent these changes in lipid profile. Objective: Verify the effects of HRT consisting of transdermal estradiol gel associated with medroxyprogesterone acetate on the lipid profile and biochemical parameters in Brazilian postmenopausal women. Methods: This study is an open prospective longitudinal study, in which thirty postmenopausal women received transdermal estradiol gel (1 mg/day) continuously combined with oral medroxyprogesterone acetate (MPA) (5 mg/day) for 12 days/month. The following parameters were determined: total cholesterol, triglycerides, high density lipoprotein (HDL), low density lipoprotein (LDL), very low density lipoprotein (VLDL), glucose, aspartate aminotransferase (AST), alanine aminotransferase (ALT), gama glutamyl transferase (GGT) and follicle-stimulating hormone (FSH). Results: The parameters of the lipid profile did not show a significant decrease, while the levels of GGT and FSH had a statistically significant decrease. Conclusions: the treatment with transdermal estradiol gel did not have a significant impact on the lipid profile, thus not resulting in a beneficial effect on cardiovascular disease markers, suggesting that the dose, administration route and the time of treatment were important for these results. Moreover, the treatment using small dose and the transdermal administration route also had a significant hepatic effect in this population. Therefore, this treatment might provide interesting effects on the lipid profile in Brazilian postmenopausal women.

Fundamento: Cambios significantes ocurren en las mujeres posmenopáusicas que pueden inducir enfermedades cardiovasculares, tales como el perfil lipídico aterogénico debido a un aumento en los niveles de colesterol total y LDL y una disminución en los niveles de HDL. La terapia de reemplazo hormonal (TRH) puede evitar esos cambios en el perfil lipídico. Objetivo: Determinar los efectos de la TRH constituida por estradiol transdérmico y acetato de medroxiprogesterona en los parámetros bioquímicos y lipídicos de mujeres brasileñas posmenopáusicas Métodos: Este es un estudio prospectivo, longitudinal, abierto, en el que treinta mujeres posmenopáusicas recibieron estradiol en gel transdérmico (1 mg/día) de forma continua, combinado con acetato de medroxiprogesterona (MPA) (5 mg/día) por 12 días/mes. Se determinaron los seguientes parámetros: colesterol total, triglicéridos, lipoproteína de alta densidad (HDL-colesterol), lipoproteína de baja densidad (LDL-colesterol), lipoproteína de muy baja densidad (VLDL-colesterol), glucosa, aspartato transaminasa (AST), alanina aminotransferasa (ALT), Gammaglutamiltranspeptidasa (GGT) y hormona foliculoestimulante (FSH). Resultados: Los parámetros del perfil lipídico mostraron una disminución insignificante, mientras los niveles de GGT y FSH presentaron una disminución estadísticamente significante. Conclusiones: El tratamiento con estradiol en gel transdérmico no mostró un impacto significante en el perfil lipídico, causando un efecto benéfico en los marcadores de enfermedades cardiovasculares, sugiriendo que la dosis, el modo de administración y el tiempo de tratamiento fueron importantes para esos resultados. Además, el tratamiento con dosis baja y modo de administración transdérmico también demostró un significante efecto hepático en esa población. Así pues, ese tratamiento puede surtir efectos interesantes sobre el perfil lipídico en las mujeres brasileñas posmenopáusicas.

Screening for Y-chromosome microdeletions in infertile Indian males: utility of simplified multiplex PCR.

BACKGROUND & OBJECTIVE: Analysis of the microdeletions in the azoospermia factor (AZF) region of Y chromosome by PCR is an important screening tool in the work-up of infertile males opting for assisted reproductive techniques. In the present study, the Y chromosome microdeletions were analyzed by PCR using primers corresponding to 16 sequence tagged sites (STS) and three genes of the AZF region in infertile Indian men. Feasibility of developing a simplified multiplex PCR for screening of the Y chromosome microdeletions has been explored. METHODS: A total of 271 male subjects were analyzed, of which, 170 were infertile patients (51 oligospermic and 119 azoospermic) and 101 were fertile controls. Subjects showing normal karyotype only were included in the study. The semen analysis was done and plasma follicle stimulating hormone (FSH) concentrations were determined by radioimmunoassay. Testicular histopathology was analyzed by fine needle aspiration cytology (FNAC). RESULTS: Y chromosome microdeletions were observed in nine out of 170 (5.29%) infertile males all of whom were azoospermic. Of the nine subjects, two had deletions in AZFa, one in AZFb, three in AZFc and three in AZFb+c regions. No deletions were observed in the infertile severe oligospermic men (< 5 million sperm/ml semen) and fertile controls. No difference in the FSH concentrations of infertile patients with and without deletions (18.36 and 18.10 mIU/ml respectively) was observed. A clear relationship between Y chromosome microdeletions and testicular phenotypes could not be established. Two multiplex PCRs were designed using 7 STSs markers, which could detect Y chromosome microdeletions in infertile male subjects as efficiently as PCR based on larger number of PCR reactions. INTERPRETATION & CONCLUSION: The multiplex PCRs described in the present study may be a suitable, cost-effective and less time consuming method for screening the Y chromosome deletions in infertile males in routine clinical diagnosis and counselling prior to assisted reproduction.

FSH, the neglected sibling: evidence for its role in regulation of spermatogenesis and Leydig cell function.

The role of follicle stimulating harmone(FSH) in male reproductive function remains a matter of debate although recent evidences strongly suggest a role despite the controversies that arose following the results obtained with FSH-beta null mice and observations from human FSH receptor mutations. This review summarizes the recent developments of our understanding on the role of FSH in male reproduction. Specifically the results obtained with FSH-beta and FORKO null mice are be discussed in light of our observations employing active and passive neutralization of endogenous FSH in rodents and primates along with other studies. On the basis of results obtained employing a variety of models it can be conclude unequivocally that FSH regulates Leydig cell function and is essential for normal quantitative spermatogenesis.

Effect of simultaneous exposure to lead and cadmium on gonadotropin binding and steroidogenesis on granulosa cells: an in vitro study.

Effects of lead (Pb) and cadmium (Cd) both alone or in combination on the binding of LH and FSH on isolated granulosa cells were studied. Granulosa cells isolated from proestrous rats were incubated (in vitro) with lead acetate and/or cadmium acetate (0.03 microM of Pb or Cd) for 1 hr. LH binding was dropped to 84% in Pb treated cells, 72.5% in Cd treated cells and 74.8% in combined metal treated cells compared to control. FSH binding dropped to 85.5% in Pb treated cells, 71.16% in Cd treated cells and 72.5% in combined metal treated cells compared to control. Activity of 17beta Hydroxy Steroid Dehydrogenase (17betaHSDH), a key steroidogenic enzyme was reduced by 52% in Cd and 37% in combined metal exposed cells whereas Pb exposed cells showed 31% reduction in the enzyme activity. Pretreatment with SH groups protectants (glutathione [GSH], dithiothretol [DTT]) and zinc caused an ameriolation in enzyme activity whereas Zn pretreatment showed an increase in gonadotropin binding in metal exposed cells. These results suggest that both Pb and Cd can cause a reduction in LH and FSH binding, which significantly alters steroid production in vitro and exerts a direct influence on granulosa cell function.

G769A Variation of Inhibin alpha -gene in Korean Women with Premature Ovarian Failure

Premature ovarian failure (POF) is menopause before the age of 40 years. The frequency of POF is about 1% of all women. Recently inhibin alpha gene (INHalpha) has been indicated as candidate in POF pathogenesis. Inhibin, a glycoprotein, is a gonadal hormone, which can inhibit the synthesis and secretion of pituitary follicle-stimulating hormone (FSH), which has an important role in the recruitment and development of ovarian follicles during the folliculogenesis. G769A variation of INH alpha, alanine, is highly conserved across species, and has an important role of its receptor binding. We screened a G769A transition in the INHalpha from the total population of the patients of 84 women with POF and 100 normal fertile women. We found no variation between the normal subjects and the POF patients. G769A variation of INHalpha is rare in Korea women with POF.

Effect of leptin on LH and FSH release in ovariectomized rats

We compared the estradiol/progesterone-induced luteinizing hormone [LH] and follicle-stimulating hormone [FSH] release between normally fed and leptin-supplemented starved ovariectomized female rats and studied also the effect of hyper-leptinaemia on the steroid-induced hormonal release in normally fed ovariectomized rats. Three days' starvation completely abolished steroid-induced LH and FSH release. Significant recovery of the hormonal release was shown in the leptin-supplemented starved group. The magnitudes of LH and FSH release in the normally fed animals with a higher dose of leptin were statistically the same as those in the normally fed group without leptin. These observations indicate that physiological concentrations of circulating leptin exert a stimulatory effect on steroid-induced LH and FSH release

Effect of naloxone on GnRH-induced LH and FSH release in buffalo, Bubalus bubalis.

The effect of naloxone on GnRH-induced LH and FSH release was measured in buffaloes in luteal phase of estrous cycle. Animals were administered intravenously, naloxone/saline (50 mg/injection) every 15 min for 3 hr followed by GnRH (100 micrograms). Peripheral plasma LH and FSH concentrations were measured in blood samples collected at 15 min intervals from 1 hr prior to beginning of naloxone/saline treatment up to 3 hr post GnRH administration and every 30 min for the subsequent 3.5 hr. Between the animals of Group I administered naloxone and those of Group II given saline, GnRH-induced peak LH and FSH concentrations, the total LH and FSH released in response to GnRH, and the time to peak LH and FSH concentrations were not significantly different. The results of the present study suggest the absence of a direct effect of naloxone on pituitary responsiveness to GnRH.

Subpopulations of physiological and ovarian follicular fluid peptide induced apoptotic cells.

Ovarian follicular fluid peptide (OFFP) purified from sheep ovaries enhances apoptotic changes in ovarian granulosa cells of mice. To get an insight into the cell subpopulations responding to OFFP, the heterogeneity of granulosa cells was resolved. Subpopulations of granulosa cells were obtained from ovaries of immature mice treated with PMSG alone and autopsied 48 hr (control) and 72 hr after injection (atretic) and from animals injected OFFP 24 hr after PMSG injection and autopsied 24 hr later (OFFP treated) by separation on discontinuous Percoll gradient. Four fractions were collected and studied for their relative distributions and percent apoptotic cells measured by acridine orange staining. FSH binding to granulosa cell (sedimenting as a major) fraction was studied by radio receptor assay. There is a difference in densities in subpopulations of apoptotic cells induced by OFFP and those generated during the physiological process of atresia. This difference may be a reflection of different granulosa cell subpopulations involved in peptide response or differences in phases as the cells transit from normal to apoptotic phenotype. FSH binding to granulosa cells from OFFP treated animals was significantly less than those from control and atretic group.

Antisense mRNA for NPY-Y1 receptor in the medial preoptic area increases prolactin secretion

We investigated the participation of neuropeptide Y-Y1 receptors within the medial preoptic area in luteinizing hormone, follicle-stimulating hormone and prolactin release. Four bilateral microinjections of sense (control) or antisense 18-base oligonucleotides of messenger ribonucleic acid (mRNA) (250 ng) corresponding to the NH2-terminus of the neuropeptide Y1 receptor were performed at 12-h intervals for two days into the medial preoptic area of ovariectomized Wistar rats (N = 16), weighing 180 to 200 g, treated with estrogen (50 µg) and progesterone (25 mg) two days before the experiments between 8.00 and 10:00 a.m. Blockade of Y1 receptor synthesis in the medial preoptic area by the antisense mRNA did not change plasma luteinizing hormone or follicle-stimulating hormone but did increase prolactin from 19.6 + or - 5.9 ng/ml in the sense group to 52.9 + or - 9.6 ng/ml in the antisense group. The plasma hormones were measured by radioimmunoassay and the values are reported as mean + or - SEM. These data suggest that endogenous neuropeptide Y in the medial preoptic area has an inhibitory action on prolactin secretion through Y1 receptors

Possible involvement of A1 receptors in the inhibition of gonadotropin secretion induced by adenosine in rat hemipituitaries in vitro

We investigated the participation of A1 or A2 receptors in the gonadotrope and their role in the regulation of LH and FSH secretion in adult rat hemipituitary preparations, using adenosine analogues. A dose-dependent inhibition of LH and FSH secretion was observed after the administration of graded doses of the R-isomer of phenylisopropyladenosine (R-PIA; 1 nM, 10 nM, 100 nM, 1 µM and 10 µM). The effect of R-PIA (10 nM) was blocked by the addition of 8-cyclopentyltheophylline (CPT), a selective A1 adenosine receptor antagonist, at the dose of 1 µM. The addition of an A2 receptor-specific agonist, 5-N-methylcarboxamidoadenosine (MECA), at the doses of 1 nM to 1 µM had no significant effect on LH or FSH secretion, suggesting the absence of this receptor subtype in the gonadotrope. However, a sharp inhibition of the basal secretion of these gonadotropins was observed after the administration of 10 µM MECA. This effect mimicked the inhibition induced by R-PIA, supporting the hypothesis of the presence of A1 receptors in the gonadotrope. R-PIA (1 nM to 1 µM) also inhibited the secretion of LH and FSH induced by phospholipase C (0.5 IU/ml) in a dose-dependent manner. These results suggest the presence of A1 receptors and the absence of A2 receptors in the gonadotrope. It is possible that the inhibition of LH and FSH secretion resulting from the activation of A1 receptors may have occurred independently of the increase in membrane phosphoinositide synthesis

A transiçäo da perimenopausa / The perimenopausal transition

Os objetivos deste artigo säo: 1- Definir idade de início e duraçäo da transiçäo da perimenopausa. 2- Descrever as alteraçöes hormonais, lipídicas e ósseas que ocorrem durante a transiçäo da perimenopausa. 3- Atentar para os principais requisitos de cuidados preventivos com a saúde no período da vida correspondente à perimenopausa.

Inhibitory activity in buffalo follicular fluid and its elution pattern during different season.

Two pools of whole buffalo follicular fluid collected in winter (December, January) and spring (March, April) season were fractionated by Sephadex G-200 column chromatography. Follicular fluid collected in winter and spring eluted into different pattern resulted into four and three peaks respectively. Both the pools of follicular fluid were salted out with 18.5% ammonium sulphate. The salted out fraction of winter and spring season follicular fluid was again subjected to sephadex column chromatography which eluted into two and single peak respectively. Whole follicular fluid (0.6 ml) was administered into ovariectomized mice to see its inhibitory effect. The percentage of compensatory ovarian hypertrophy was 16.3 +/- 4.4 which was significantly different (P < 0.01) as compared to control. 200 micrograms material from peak 1 and peak 2 obtained after fractionation of salted out winter follicular fluid also had inhibitory effect on compensatory ovarian hypertrophy as compared to control group. It was 35.6 +/- 9.3 and 15.9 +/- 4.3% respectively. Thus, the variation in nature of buffalo follicular fluid and its inhibitory effect in ovariectomised mice have significant relation with anoestrus condition in summer and humid months in this species.

Contribuiçäo para o estudo do comportamento da prolactina endometrial / Study endometrial prolactin secretion

Com objetivo de verificar o comportamento da prolactina endometrial, foram estudadas 40 mulheres, com média etaria de 28,3 anos e selecionadas por apresentarem os seguintes sintomas : alteraçäo do ciclo menstrual e/ou esterilidade e/ ou galactorreia. As pacientes foram distribuidas entre os seguintes grupos : Grupo G1, dez mulheres hiperprolactinemicas com menstruaçöes regulares ; G2, nove mulheres hiperprolactinemicas com menstruaçöes irregulares; Grupo G3, onze mulheres normoprolactinemicas com menstruaçöes regulares; Grupo G4, dez mulheres normoprolactinemicas com menstruaçöes irregulares. As 40 mulheres foram redistribuidas segundo o padrao endometrial nos seguintes grupos : Grupo G5, mulheres com ciclos ovulatorios e Grupo G6, mulheres com ciclos anovulatórios. Nas pacientes ovulatórias, ainda correlacionaram-se os percentuais de prolactina endometrial com fase lutea adequada e inadequada. Exames laboratoriais foram realizados na fase folicular inicial (do terceiro ao nono dia), fase lutea inicial (do 15. ao 21. dia) e fase lutea tardia ( do 22. ao 29. dia), nas pacientes que menstruavam (1,2 e 3 amostras, respectivamente) e, nas que näo menstruavam, no primeiro, 14 e 21 dia da consulta (1,2 e 3 amostras, respectivamente)...

Endocrinologia do ovário no climatério perimenopáusico / The ovarian endocrinology in the perimenopausal period

O climatério, período de transiçäo entre o menacme e a senectude, compreende uma série de fenômenos clínicos, biológicos e endocrinológicos, que se iniciam muito antes da instalaçäo da menopausa e prosseguem por anos após a mesma. A endocrinologia de transiçäo menopáusica, compreendida nesse período, é o objetivo do presente trabalho de revisäo da literatura. Observa-se, inicialmente, diminuiçäo na secreçäo de Inibina, um polipeptídeo produzido pelas células da granulosa, responsável por modulaçäo da secreçäo de FSH. Há aumento do FSH, seguido de aumento do LH e diminuiçäo da esteroidogênese ovariana, resultando em hipoestrogenismo e diminuiçäo dos níveis de progesterona, o que concorre para a série de alteraçöes que caracterizam esse período.