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Extracellular vesicles (EV),nanoscale vesicles encapsulated by phospholipid bilayers,are rich in biological molecules such as nucleic acids,metabolites,proteins,and lipids derived from parental cells.They are mainly involved in intercellular communication,signal transmission,and material transport and affect the functions of target cells.Ovulation disorders account for a higher proportion in the factors causing infertility which demonstrates increasing incidence year by year.Non-coding RNAs participate in a series of physiological and pathological processes of follicular development,playing a key role in female infertility.This review systematically introduces the types and biological roles of EV and elaborates on the regulation of follicular development from the effects of EV and non-coding RNAs on granulosa cell function,oocyte maturation,ovulation,luteal formation,and steroid hormone synthesis,providing a new idea and a breakthrough point for the diagnosis and treatment of infertility.
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Femenino , Humanos , Oogénesis/fisiología , Células de la Granulosa , Vesículas Extracelulares/fisiología , Comunicación Celular , ARN no Traducido , InfertilidadRESUMEN
In recent years,microRNAs(miRNAs)have been detected at different stages of follicular development and in different cells of follicles.Extracellular vesicle(EV)-derived miRNAs have also been detected in the follicular fluid of mature follicles.miRNAs participate in the regulation of normal follicular development,and the regulation disorder may lead to the occurrence of some ovarian diseases.In order to further systematically elucidate the regulatory mechanism of miRNAs on follicular development and find suitable EV-derived miRNAs that can predict oocyte development,we reviewed the functions of miRNAs in follicular development from the perspectives of granulosa cell development,oocyte development,and hormone synthesis.
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Femenino , Humanos , Líquido Folicular , Células de la Granulosa , MicroARNs/genética , Oogénesis , Folículo OváricoRESUMEN
Polycystic ovary syndrome (PCOS) is a common endocrine disease of child-bearing period women and one of the main causes of infertility in women. Pentraxin 3 (PTX3) is a multifunctional protein with a series of biological activities. PTX3 participates in the regulation of insulin secretion and glucose metabolism, ovarian cumulus cell function, inflammatory factor activity, androgen metabolism, lipid absorption and transport, and endothelial cell function, thereby improving insulin resistance, promoting follicular development and ovulation, reducing chronic inflammation, inhibiting androgen levels, improving lipid metabolism abnormalities and preventing atherosclerosis and cardiovascular diseases, thus participating in the occurrence of PCOS and its complications. This article reviews the mechanism of PTX3 in PCOS and its complications, trying to provide new ideas and directions for the study of PCOS pathogenesis and its clinical diagnosis and treatment.
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Niño , Femenino , Humanos , Proteína C-Reactiva/metabolismo , Resistencia a la Insulina , Síndrome del Ovario Poliquístico/fisiopatología , Componente Amiloide P Sérico/metabolismoRESUMEN
Extracellular vesicles (EVs) refer to bilayer membrane transport vesicles secreted by cells. EVs can take macromolecules from cells and transfer them to receptor cells. Among these macromolecular substances, the most studied are microRNAs (miRNAs). miRNA is non-coding RNA involved in the regulation of gene expression. It has been confirmed that there are different non-coding RNAs in mammalian follicular fluid EVs. EVs carrying miRNA can act as an alternative mechanism for autocrine and paracrine, affecting follicular development. This paper systematically introduced the kinds, characteristics and methods of isolation and identification of EVs, focusing on the effects of EVs and miRNAs on follicular development, including early follicular development, oocyte maturation, follicular dominance and effects on granulosa cell function. At the same time, the authors prospected the future research of EVs and microRNAs in follicular fluid, and provided ideas and directions for the research and application of EVs and miRNA functions in follicular fluid.
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Animales , Femenino , Vesículas Extracelulares , Metabolismo , Líquido Folicular , Química , Células de la Granulosa , MicroARNs , Farmacología , OogénesisRESUMEN
OBJECTIVE@#To observe the effect of acupuncture and moxibustion therapy of on ovulation and embryo implantation in luteal phase defect patients with spleen-kidney deficiency.@*METHODS@#A total of 80 patients were randomly divided into an observation group and a control group, 40 cases in each one.In the observation group,acupuncture was applied at Shenting (GV 24), Shenque (CV 8), Guanyuan (CV 4), Qixue (KI 13), Lieque (LU 7), Gongsun (SP 4), Taixi (KI 3), Zusanli (ST 36) and Taichong (LR 3). And moxibustion was given at Taixi (KI 3) using moxibustion box during follicular phase, the stimulation of Taichong (LR 3) was strengthened during ovulatory phase, moxibustion was adopted at Shenque (CV 8) to Guanyuan (CV 4), Zusanli (ST 36) and Taixi (KI 3) during luteal phase. In the control group, acupuncture was applied at Guanyuan (CV 4), Dahe (KI 12), Sanyinjiao (SP 6), Ciliao (BL 32), Zhibian (BL 54) and Shenque (CV 8). Moxibustion was given at Sanyinjiao (SP 6) using moxibustion box during follicular phase, and moxibustion was adopted at Shenque (CV 8) to Guanyuan (CV 4) during luteal phase. The treatment were given every Monday, Wednesday and Friday, and the treatment were stoped during menstrual period in the two groups. Totally 3 menstrual cycle treatment were required, and 3 menstrual cycles were followed up. The pregnancy rate was observed after treatment, the ovulation rate, maximum folliclular diameter and difference of maximum folliclular diameters in ovulatory phase, serum progesterone (P) and basal body temperature (BBT) were compared before and after treatment in the two groups.@*RESULTS@#In the observation group, 6 cases of successful pregnancy during treatment,10 cases in follow-up, the clinical pregnancy rate was 40.0% (16/40). In the control group, 1 case of successful pregnancy during treatment, 5 cases in follow-up, the clinical pregnancy rate was 15.0% (6/40). The clinical pregnancy rate in the observation group was higher than the control group (0.05). Compared before treatment, the maximum folliclular diameter and difference of maximum folliclular diameters in ovulatory phase, serum P after treatment were improved in the two groups (<0.05), and the improvements of the observation group were significant compared with the control group (<0.05). The BBT after treatment were superior to before treatment in the two groups (<0.05). After treatment, the normal BBT in the observation group was 33 cases, while the control group was 22 cases (<0.05).@*CONCLUSION@#Acupuncture and moxibustion therapy of can promote folliclar development, improve dominant follicle morphology, increase the level of serum P. The therapeutic effect is superior to routine acupuncture in increasing ovulation rate and improving pregnancy outcome.
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Femenino , Humanos , Embarazo , Puntos de Acupuntura , Terapia por Acupuntura , Infertilidad Femenina , Terapéutica , Fase Luteínica , Moxibustión , Resultado del EmbarazoRESUMEN
Studies on reproduction in sea turtles are important due to its life cycle, migratory patterns, high juvenile mortality and environmental impacts. This study aimed to analyse histomorphometrically gonads of C. mydas from the coastline of the Espírito Santo State, Brazil. Ovaries and testicles were collected between 2014 and 2015 from stranded animals. The material was fixed in formalin 10%, assessed macroscopically and processed for histomorphometrical evaluation. Gonads from 34 individuals were evaluated, twenty-four females and ten males. Macroscopic sexual identification presented 100% accuracy confirmed by histology. Sexual dimorphism was observed in one individual, which was considered as adult (CCL=1.023 m). Microscopy of female gonads revealed predominant previtellogenic follicles; oocyte diameter ranged between 161µm and 750µm and a positive correlation between ovarian length, largest oocyte and CCL was found. In males, autolysis was verified in five individuals. Viable testicles revealed predominant spermatogonia, primary spermatocytes and Sertoli cells in the seminiferous tubules and, Leydig cells and fibroblasts in the stroma. There was a positive correlation between tubular diameter and CCL and testicle length and CCL. Maturation of stromal tissue and a positive correlation between tubular lumen and CCL were also observed. Gonad development is proportional to individual growth.(AU)
Estudos em reprodução de tartarugas marinhas são importantes devido ao ciclo de vida, ao padrão migratório, à alta mortalidade juvenil e aos impactos ambientais. Objetivou-se analisar histomorfometricamente gônadas de C. mydas no litoral do Espírito Santo. Foram coletados ovários e testículos dessa espécie, entre 2014 e 2015. O material foi fixado em formol a 10% e avaliado macroscopicamente. Em seguida, foi processado para avaliação histomorfométrica. Foram avaliadas gônadas de 34 indivíduos, 24 fêmeas e 10 machos. Verificaram-se 100% de acurácia na identificação sexual à macroscopia, confirmada pela histologia. Observou-se dimorfismo sexual em um macho, que foi considerado adulto (CCC=1,023m). A microscopia dos ovários revelou folículos pré-vitelogênicos, cujos ovócitos apresentaram diâmetro médio entre 161µm e 750µm. Houve correlação positiva entre comprimento ovariano e diâmetro do maior ovócito e CCC. Nos machos, verificou-se autólise em cinco indivíduos. Os testículos viáveis revelaram espermatogônias, espermatócitos primários e células de Sertoli nos túbulos seminíferos, além de células de Leydig e fibroblastos no estroma. Houve correlação positiva entre diâmetro tubular e CCC e comprimento testicular e CCC. Verificou-se maturação do tecido estromal e correlação positiva entre o diâmetro do lúmen tubular e o CCC. Verifica-se que o desenvolvimento das gônadas é proporcional ao crescimento dos indivíduos.(AU)
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Animales , Testículo/crecimiento & desarrollo , Tortugas/anatomía & histología , Gónadas/anomalías , HistologíaRESUMEN
Polycystic ovary syndrome (PCOS) is the most common endocrine disorder affecting women of childbearing age, which features oligo- or anovulation, polycystic ovaries, hyperandrogenism and the related clinical signs, such as acne and hirsutism. At present, PCOS patients are considered to be in a long-time condition of chronic inflammation. It is reported that increased expression of inflammatory factors and/or increased levels of inflammation exist in peripheral blood, granulosa cells, follicular fluid, ovarian stroma, adipocytes and endometrial cells in patients with PCOS. Studies on the role of inflammatory factors in the pathogenesis of PCOS suggest that inflammatory factors may have an influence on the clinical outcome through affecting follicular development, androgen levels and so on.
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<p><b>OBJECTIVE</b>To evaluate the effect of Heyan Kuntai Capsule (, HYKT) on the ovarian function of aged mice and expressions of cohesion complexes in oocytes.</p><p><b>METHODS</b>Twenty-five 9-month-old female C57BL/6J mice were randomly divided into 5 groups by block randomization method (n=5 per group), including the control group (saline), 17β-estradiol group [E, 100 μg/(kg•d)], and low-, medium-, and highdose of HYKT groups [0.3, 0.9, 2.7 g/(kg•d), respectively]. All mice were treated by intragastric administration for 4 weeks. Hematoxylin and eosin staining and anti-VASA staining were used to detect the amounts of follicles. The apoptosis of follicles was measured by anti-gamma HA histone family member X (γHAX) staining and TdT-mediated dUTP Nick-End Labeling (TUNEL) assay. The density of cohesin subunits, REC8 meiotic recombination protein (REC8), structural maintenance of chromosome (SMC) 1β and SMC3 in oocytes were evaluated by immunofluorescent staining.</p><p><b>RESULTS</b>After administration of E and high-dose of HYKT, the total number of follicles as well as the number of primordial and primary follicles were significantly increased (P<0.05). Anti-γHAX staining and TUNEL assay demonstrated that high-dose of HYKT and E partly suppressed the apoptosis of follicles (P<0.05). Furthermore, it showed an increased trend in the levels of REC8 and SMC1β, after administration with E and HYKT, and no obvious change in the level of SMC3.</p><p><b>CONCLUSION</b>HYKT could enhance the number of follicles, suppress apoptosis of oocytes and have a trend to elevate the meiotic-specific cohesin subunits (REC8 and SMC1β) in oocytes of aged mice, indicating a beneficial effect on the ovarian function in terms of the quantity and quality of follicles.</p>
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In the theory of traditional Chinese medicine(TCM) that "kidney storing essence and governing reproduction", reproductive essence is an important part of the kidney essence and acts as the original material of offspring embryos. Sperm, oocyte and zygote should be all included in the range of reproductive essence. Ovum is the essence of reproduction from inborn. The follicles maturation depends on the quality of oocyte and the vigor of kidney essence. Meanwhile, discharge of mature ovum relies on the stimulation and promotion by kidney Qi. Autophagy almost exists in different cells stages and all various of mammalian cells. Many studies have found that autophagy not only participates in the formation of follicles, but also in every phase of the follicles development, and is involved in the occurrence and development of ovarian diseases. Recently, more and more scholars believe that autophagy is a new field to explore the microcosmic relationship between autophagy and TCM. Kidney-nourishing TCM could promote follicular growth and improve variety clinical symptoms by inhibiting the apoptosis of ovarian granulosa cells and reducing follicular atresia. Meanwhile, apoptosis of ovarian granulosa cells is closely related to autophagy of ovarian granulosa cells. In order to provide some theoretical foundation for kidney-nourishing therapy's promoting effect on follicular growth and improving effect on ovarian function, also to further explore the molecular mechanism of kidney-nourishing medicine in promoting follicular development, this paper would explain the microcosmic relationship between autophagy and follicular development based on the theory of "kidney governing reproduction". All of these would be of great significance to prevent and intervene the diseases of reproductive system timely and effectively.
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Polycystic ovary syndrome (PCOS) is the most common endocrine disorder affecting women of childbearing age, which features oligo-or anovulation, polycystic ovaries, hyperandrogenism and the related clinical signs, such as acne and hirsutism. At present, PCOS patients are considered to be in a long-time condition of chronic inflammation. It is reported that increased expression of inflammatory factors and/or increased levels of inflammation exist in peripheral blood, granulosa cells, follicular fluid, ovarian stroma, adipocytes and endometrial cells in patients with PCOS. Studies on the role of inflammatory factors in the pathogenesis of PCOS suggest that inflammatory factors may have an influence on the clinical outcome through affecting follicular development, androgen levels and so on.
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Background: Ornithine decarboxylase antizyme 1 (OAZ1) is an important regulator of polyamine synthesis and uptake. Our previous studies indicated that high OAZ1 expression in the ovaries of laying geese is responsible for poor egg production. In the present study, the molecular characterization of goose OAZ1 gene was analyzed, as well as the expression profile in various follicular tissues. Results: An 873-bp cDNA sequence of the OAZ1 gene (Accession No. KC845302) with a +1 frameshift site (+175T) was obtained. The sequence consisted of a 652-bp two overlapping open reading frames (a putative protein with 216 amino acids). The OAZ domain, OAZ signature and OAZ super family domain were prominent conserved regions among species. As the follicle size increased, OAZ1 abundance showed an increasing trend during follicular development, while it decreased during follicular regression. The level of OAZ1 mRNA expression was the lowest in the fifth largest preovulatory follicle, and was 0.65-fold compared to the small white follicle (P b 0.05). OAZ1 mRNA expression in the largest preovulatory and postovulatory follicle was 2.11- and 2.49-fold compared to the small white follicle, respectively (P b 0.05). Conclusions: The goose OAZ1 structure confirms that OAZ1 plays an important role in ornithine decarboxylase-mediated regulation of polyamine homeostasis. Our findings provide an evidence for a potential function of OAZ1 in follicular development, ovulation and regression.
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Animales , Femenino , Proteínas/genética , Proteínas/metabolismo , Gansos/metabolismo , Folículo Ovárico/metabolismo , Ornitina Descarboxilasa/metabolismo , Poliaminas/metabolismo , ARN Mensajero , Clonación Molecular , Análisis de Secuencia , ADN Complementario , Reacción en Cadena en Tiempo Real de la Polimerasa , Folículo Ovárico/crecimiento & desarrolloRESUMEN
AIM:To investigate the expression and localization of autophagy related protein microtublule associated protein 1 light chain 3 (LC3) at various stages of follicular development and atresia in the mice.METHODS:On 0,1,2,3,4 and 5 day after intraperitoneal injection of pregnant mare serum gonadotropin (PMSG),expression and positioning situation of autophagy related protein LC3 and apoptosis related protein cleaved caspase-3 were examined by the method of immunohistochemical staining.The protein levels of cleaved caspase-3 and LC3 were determined by Western blot in cultured mouse granulosa cells after incubation under serum-free conditions in the absence or presence of FSH.LC3 subcellular localization in granulosa cells were studied by the method of immunofluorescence.RESULTS:The LC3 protein expressed in granulosa cells during all developmental stages mainly.Granulosa cells of atretic follicles that showed intense staining of cleaved caspase-3 and LC3.The protein levels of cleaved caspase-3 and LC3-Ⅱ in the granulosa cells significantly decreased at 1 d and 2 d after intraperitoneal injection of PMSG (P < 0.05).The protein levels of cleaved caspase3 and LC3-Ⅱ in the granulosa cells increased in turn on 3,4 and 5 day after intraperitoneal injection of PMSG.The positive correlation between LC3-Ⅱ and cleaved caspase-3 protein levels was observed (r2 =0.8299,P < 0.05).The LC3-Ⅱ protein expressed with punctuate structures in granulosa cell cytoplasm cultured under serum-free conditions in the presence of FSH.CONCLUSION:LC3 is expressed in the follicular granulosa cells with cell specificity and regional specificity.Autophagy is induced mainly in granulosa cells during folliculogenesis and shows positive correlation with apoptosis.Ovarian granulosa cell autophagy and apoptosis are gonadotropic hormone dependent.
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AIM:To investigate the expression and localization of autophagy related protein microtublule associated protein 1 light chain 3 (LC3) at various stages of follicular development and atresia in the mice.METHODS:On 0,1,2,3,4 and 5 day after intraperitoneal injection of pregnant mare serum gonadotropin (PMSG),expression and positioning situation of autophagy related protein LC3 and apoptosis related protein cleaved caspase-3 were examined by the method of immunohistochemical staining.The protein levels of cleaved caspase-3 and LC3 were determined by Western blot in cultured mouse granulosa cells after incubation under serum-free conditions in the absence or presence of FSH.LC3 subcellular localization in granulosa cells were studied by the method of immunofluorescence.RESULTS:The LC3 protein expressed in granulosa cells during all developmental stages mainly.Granulosa cells of atretic follicles that showed intense staining of cleaved caspase-3 and LC3.The protein levels of cleaved caspase-3 and LC3-Ⅱ in the granulosa cells significantly decreased at 1 d and 2 d after intraperitoneal injection of PMSG (P < 0.05).The protein levels of cleaved caspase3 and LC3-Ⅱ in the granulosa cells increased in turn on 3,4 and 5 day after intraperitoneal injection of PMSG.The positive correlation between LC3-Ⅱ and cleaved caspase-3 protein levels was observed (r2 =0.8299,P < 0.05).The LC3-Ⅱ protein expressed with punctuate structures in granulosa cell cytoplasm cultured under serum-free conditions in the presence of FSH.CONCLUSION:LC3 is expressed in the follicular granulosa cells with cell specificity and regional specificity.Autophagy is induced mainly in granulosa cells during folliculogenesis and shows positive correlation with apoptosis.Ovarian granulosa cell autophagy and apoptosis are gonadotropic hormone dependent.
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The study aimed to quantify the concentrations of free IGF-I in serum and fluid of ovarian follicles in pre-pubertal gilts and describe the ovarian morphology by measuring the size of the ovaries and counting the number of surface follicles. Ovaries (n=1,000) from pre-pubertal gilts were obtained immediately after slaughter. A total of 10 samplings were performed, with ovaries obtained from 50 females for each collection. The follicles situated on the surface of each ovary were classified as small (SFs, 2 to 5mm in diameter) or large (LFs 6 to 10mm in diameter) and the follicular fluid was obtained by follicle aspiration. The collection of serum samples was performed after the gilts exsanguination using sterile tubes. From the pool of serum and follicular fluid obtained from 50 females, the concentration of free IGF-I was determined in each sample using an enzyme immunoassay kit (ELISA). The description of ovarian morphometry was performed in 100 ovaries from randomly selected gilts. The larger and smaller lengths of ovaries were measured, and the total number of SFs and LFs present on the surface of each ovary were also counted. The IGF-I concentration was greater (P<0.05) in LFs (170.92±88.29 ng/mL) compared with SFs (67.39±49.90ng/mL) and serum (73.48±34.63ng/mL). The largest and smallest length of the ovaries was 26.0±3.0 and 19.0mm ±2.0mm, respectively. The number of SFs (70.86±25.76) was greater (P<0.01) than LFs (6.54±5.26). The study concluded that LFs present greater levels of IGF-I when compared with SFs and blood, which is related to increased activity of the LFs and its differentiation to ovulation. In addition, ovaries of pre-pubertal gilts have a higher number of SFs compared to LFs. Therefore, our study demonstrated unique data regarding the physiological concentration of free IGF-I in ovarian follicles, that can be used in future research to evaluate the addition of this hormone in the in vitro production media of porcine embryos with the...
Objetivou-se quantificar as concentrações do IGF-I livre no soro e no fluido de folículos ovarianos de fêmeas suínas pré-púberes e descrever a morfologia ovariana, por meio da mensuração das dimensões dos ovários e da contagem do número de folículos superficiais. Ovários (n=1.000) foram obtidos de fêmeas pré-púberes imediatamente após o abate. Foi realizado um total de 10 coletas, sendo em cada, obtidos ovários de 50 fêmeas. Os folículos localizados na superfície de cada ovário foram classificados em pequenos (FPs, 2-5mm de diâmetro) ou grandes (FGs, 6-10mm de diâmetro) e o fluido folicular foi obtido por aspiração dos folículos. A coleta do soro foi realizada após a exsanguinação das fêmeas com o uso de tubos estéreis. A partir do pool de fluido folicular e do soro obtido das 50 fêmeas, determinou-se a concentração de IGF-I livre em cada amostra por meio de kit de ensaio imunoenzimático (ELISA). A descrição da morfometria ovariana foi realizada em 100 ovários provenientes de fêmeas escolhidas aleatoriamente. Foi mensurado o comprimento maior e menor dos ovários e, também, contabilizado o número total de FPs e FGs presentes na superfície de cada ovário. A concentração de IGF-I foi superior (P<0,05) nos FGs (170,92±88,29ng/mL) em comparação com os FPs (67,39±49,90ng/mL) e o sérico (73,48±34,63ng/mL). O comprimento maior e menor dos ovários foi de 26,0±3,0mm e 19,0±2,0 mm, respectivamente. O número de FPs (70,86±25,76) foi maior (P<0,01) em comparação com os FGs (6,54±5,26). Conclui-se que FGs apresentam níveis de IGF-I superiores aos FPs, e ao sangue, sendo isso relacionado a maior atividade dos FGs e à diferenciação que os mesmos sofrem para a ovulação. Além disso, ovários de fêmeas suínas pré-púberes apresentam elevado número de FPs em comparação aos FGs. Portanto, nosso estudo demonstrou dados originais a respeito da concentração fisiológica de IGF-I livre em folículos ovarianos, que podem ser utilizados em futuras pesquisas para avaliar a adição de...
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Animales , Femenino , Factor I del Crecimiento Similar a la Insulina/efectos adversos , Folículo Ovárico , Porcinos/crecimiento & desarrollo , Ensayo de Inmunoadsorción Enzimática/veterinariaRESUMEN
This study was aimed to observe the therapeutic effect of Bushen Tiaogan of traditional Chinese medicine (TCM) herbs combined with clomiphene citrate therapy of promoting small follicular development. A total of 70 small follicle ovulation infertility cases were randomly divided into the control group of 35 cases with the treatment of clomiphene citrate of 50 to 100 mg for 5 days, and 35 cases in the treatment group with TCM herbal medicine of Radix Polygoni Multiflori, Lycium c hinense , eucommia, bupleurum root, turmeric, Fructus aurantii to nourish the kidney and regulate the liver. The clinical efficacy and maximum follicular diameter (MFD) were observed. The results showed that the cycle pregnancy rate in the treatment group was 43 . 59%, and 35 . 13% in the control group. The curative effect of treatment group was obviously superior to the control group. There was statistical significance in the curative effect ( P < 0 . 05 ) . The incidence of LUFS was 7 . 69% in the treatment group , and 29 . 73% in the control group . There was statistical significance ( P < 0 . 01 ) . The MFD value ( 18-28 mm ) in the ovulation cycle, the cycle pregnancy rate was 44.44% in the control group and 55.17% in the treatment group. There was statistical significance difference ( P < 0 . 05 ) . The early abortion rate was 18 . 75% in the control group and 6.25% in the treatment group. There was statistical significance difference (P < 0.01). It was con-cluded that Bushen Shugan Y angyin TCM herbs combined with clomiphene citrate can promote small follicular development with obvious clinical curative effect .
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Administration of 5 mg methomyl (40%) kg-1 b.wt. mouse-1 (equal to 50% of LD50 dose), every day for 90 days to adult female mice resulted in a significant decrease in body weight, relative weight of the ovary, uterus and fallopian tube; mean number of small, preantral, antral and pre-ovulatory follicles and fertility compared to controls. On the other hand, total duration of the estrous cycle was significantly increased compared to controls. One month after the cessation of the treatment (a commercial methomyl preparation-lannate) the effect on estrous cycle and organ weight was not restored. Treatment of 2.5 mg or 1 mg lannate kg-1 b. wt., although did not alter duration of the estrous cycle; relative weight of the ovary, uterus, and fallopian tube and fertility, caused a significant decrease in mean number of small follicles compared to controls. All the groups of mice treated with lannate showed loss in body weight (15.15% in 1 mg, 6.61% in 2 mg and 12.16% in 5 mg treated groups) whereas controls showed a gain in body weight (20.02%) during the period of experimentation. The results indicate that 5 mg lannate kg-1 b. wt. causes loss of follicles and infertility, whereas lower dosages (2.5 and 1 mg) reduce the number of small follicles which might shorten reproductive life span of mice.
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The effect of season and body condition on the follicular development and the ovulation rate in Pelibuey ewes were assessed. During the periods August-November and February-May, 38 ewes were distributed in two groups according to their body condition score: high body condition (HBC) and low body condition (LBC). Estrus was detected twice a day and the ovaries were daily examined by ultrasonography through an interestral period. Ovulation rate (OR) was determined by ultrasonography and then confirmed by laparoscopy. An effect of body condition on the estrous length was found (29.6 ± 2.3 and 20.2 ± 2.5 h HBC and LBC, respectively), whereas there were no differences in the estrous cycle length and the maximum follicular diameter. The number of follicles > 4 mm and the ovulation rate were higher in ewes of high body condition and during the main reproductive season (1.8 vs 1.3 and 2.0 vs 1.4 OR and follicles, respectively).
Se evaluó el efecto del periodo del año y de la condición corporal sobre el desarrollo folicular y la tasa ovulatoria en ovejas de la raza Pelibuey. En los periodos de agosto-noviembre y febrero-mayo, 38 ovejas fueron distribuidas en dos grupos de acuerdo con su condición corporal: condición corporal alta (CCA) y condición corporal baja (CCB). Se trabajó durante dos veces al día para detectar estros, además los ovarios de las ovejas fueron examinados diariamente mediante ultrasonografía durante un periodo interestral. La tasa ovulatoria se determinó con ultrasonografía y se confirmó por medio de laparoscopía. Se encontró efecto de la condición corporal sobre la duración del estro (29.6 ± 2.3 y 20.2 ± 2.5 h CCA y CCB, respectivamente), mientras que no se observaron diferencias en la duración del ciclo estral y el diámetro folicular máximo. La tasa ovulatoria y el número de folículos > 4 mm fue mayor en las ovejas de CCA y en la época de mayor actividad reproductiva (1.8 vs 1.3 y 2.0 vs 1.4 TO y folículos, respectivamente).
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Mesenchymal stem cells (MSCs) secrete a variety of cytokines and growth factors in addition to self-renewal and multiple forms of differentiation. Some of these secreted bioactive factors could improve meiotic maturation in vitro and subsequent embryo developmental potential. The aim of the present study was to determine whether in vitro maturation (IVM) of mouse oocyte with or without cumulus cells could be improved by contact with conditioned medium (CM) of MSCs as well as the efficiency of CM to support follicular growth and oocyte maturation in the ovarian organ of mice cultured on soft agar. The developmental potential of matured oocyte was assessed by blastocyst formation after in vitro fertilization (IVF). Germinal vesicle stage oocytes with or without cumulus cells were subjected to IVM in either CM, Dulbecco's modified Eagle's medium (DMEM), alpha-minimum essential medium (alpha-MEM) or human tubal fluid (HTF). Approximately 120 oocytes were studied for each medium. CM produced a higher maturation rate (91.2%) than DMEM (54.7%), alpha-MEM (63.5%) and HTF (27.1%). Moreover, CM improved embryo development to blastocyst stage significantly more than DMEM and HTF (85 vs 7% and 41.7%, respectively) but there was no significant difference compared with alpha-MEM (85 vs 80.3%). The behavior of cortical granules of IVM oocytes cultured in CM revealed cytoplasmic maturation. Moreover, CM also supported preantral follicles growth well in organotypic culture on soft agar resulting in the maturation of 60% of them to developmentally competent oocytes. The production of estrogen progressively increased approximately 1-fold every other day during organ culture, while a dramatic 10-fold increase in progesterone was observed 17 h after human chorionic gonadotropin stimulus at the end of culture. Thus, CM is an effective medium for preantral follicle growth, oocyte maturation, and sequential embryo development.
Asunto(s)
Animales , Femenino , Ratones , Embarazo , Medios de Cultivo Condicionados/farmacología , Células Madre Mesenquimatosas , Oocitos/crecimiento & desarrollo , Células del Cúmulo/citología , Embrión de Mamíferos/embriología , Desarrollo Embrionario/fisiología , Fertilización In Vitro , Meiosis/fisiología , Folículo Ovárico/crecimiento & desarrolloRESUMEN
OBJECTIVE: Identification of the regulatory mechanism for arrest and initiation of primordial follicular growth is crucial for female fertility. Previously, we found 15 expressed sequence tags (ESTs) that were specifically abundant in the day-5-subtracted cDNA library and that the B357 clone was novel. The present study was conducted to obtain the whole sequence of the novel gene including B357 and to characterize its mRNA and protein expression in mouse ovary and testis. METHODS: The extended sequence of the 2,965-bp cDNA fragment for the clone B357 was named 5-day-ovary-specific gene-1 (5DOS1) and submitted to GenBank (accession number AY751521). Expression of 5DOS1 was characterized in both female and male gonads at various developmental stages by Northern blotting, real-time RT-PCR, in situ hybridization, Western blotting, and immunohistochemistry. RESULTS: The 5DOS1 transcript was highly expressed in the adult testis, brain, and muscle as compared to the other tissues. In the ovary, the 5DOS1 transcript was detected in all oocytes from primordial to antral follicles, and highly expressed at day 5 after birth and decreased thereafter. In contrast, expression of 5DOS1 showed a gradual increase during testicular development and its expression was limited to various stages of male germ cells except spermatogonia. CONCLUSIONS: This is the first report on the expression and characterization of the 5DOS1 gene in the mouse gonads. Further functional analysis of the 5DOS1 protein will be required to predict its role in gametogenesis.
Asunto(s)
Adulto , Animales , Femenino , Humanos , Masculino , Ratones , Northern Blotting , Western Blotting , Encéfalo , Células Clonales , Análisis por Conglomerados , Bases de Datos de Ácidos Nucleicos , ADN Complementario , Etiquetas de Secuencia Expresada , Fertilidad , Gametogénesis , Biblioteca de Genes , Células Germinativas , Gónadas , Inmunohistoquímica , Hibridación in Situ , Oocitos , Ovario , Parto , ARN Mensajero , Espermatogonias , TestículoRESUMEN
0.05).There were more polycystic ovary (PCO) and (or) polycystic ovarian syndrome (PCOS) patients,more basal antra] follicles,longer duration of Gn stimulation (range 16-33 days),higher Gn dose,lower serum peak estradiol (E_2) level,fewer oocytes,fewer embryos transferred,in group 1 compared with group 2 (P