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1.
Rev Assoc Med Bras (1992) ; 65(8): 1122-1127, 2019 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-31531613

RESUMO

Melatonin is known for its effects on both the sleep and reproductive system of mammals. The latter has melatonin receptors type 1 and 2, which act to regulate, among other things, cyclic AMP. Notwithstanding all the literature data, there is still no sound knowledge or a clear understanding of the hormone's action on the physiology of ovarian follicular cells. OBJECTIVE To review and evaluate studies about melatonin action on the ovarian granulosa/theca interna cells from the literature. METHODS The systematic review was carried out according to the PRISMA recommendations. The MEDLINE and Cochrane primary databases were consulted with the use of specific terms. There was no limitation on language or publication year. RESULTS Seven papers about melatonin action on granulosa cells were selected. The following can be attributed to the hormone's effects: a) progesterone increase in culture medium; b) increased estrogen production; c) antagonistic action on estrogen; d) improvement in cell quality resulting in improved embryo and higher pregnancy rates; e) improved cell proliferation via MAPK; f) reduction of free radicals. Nevertheless, there are contrarian papers reporting a reduction in progesterone production. Melatonin interferes in sex steroid production, boosting progesterone output. Such action may help improve oocyte quality.


Assuntos
Melatonina/farmacologia , Oócitos/efeitos dos fármacos , Folículo Ovariano/efeitos dos fármacos , Células Cultivadas , Feminino , Células da Granulosa/efeitos dos fármacos , Humanos , Oócitos/crescimento & desenvolvimento , Gravidez , Progesterona/antagonistas & inibidores , Células Tecais/efeitos dos fármacos
2.
Life Sci ; 235: 116810, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31472147

RESUMO

AIMS: Previous reports have demonstrated that melatonin exists in multiple extrapineal sites, and higher amounts of melatonin are present in human follicular fluid than in serum, which indicates that it might play key roles in human oocyte maturation and subsequent embryonic development. Melatonin has been shown to be a potent antioxidant and might be beneficial to human oocytes during in vitro maturation (IVM). However, the underlying mechanisms of melatonin action during IVM have not been thoroughly investigated. MAIN METHODS: Immunofluorescence staining, western blotting, and ELISA were applied to investigate whether melatoninergic components are expressed in the cultured human ovarian cumulus cells. TMRE staining and Fluo-4 AM staining were performed to detect the mitochondrial membrane potential and intracellular Ca2+ levels of immature human oocytes respectively. KEY FINDINGS: First, cultured human ovary cumulus cells synthesized melatonin in vitro, and it expressed serotonin (the precursor of melatonin) and the two key enzymes, i.e. N-acetyltransferase (NAT) and hydroxyindole-O-methyltransferase (HIOMT). Additionally, the results suggest that melatonin maintains the mitochondrial membrane potential and decrease excessive Ca2+ levels in immature human oocytes during IVM. SIGNIFICANCE: In conclusion, we provide evidence that the melatoninergic components were expressed in cultured human ovarian cumulus cells, and melatonin might reduce oxidative stress of human oocytes by ameliorating mitochondrial function. In view of the significant clinical value that immature human oocytes have in assisted reproductive technology (ART), our findings highlight a potential treatment strategy of using melatonin to improve mitochondrial function and to enhance the quality of human oocytes during IVM.


Assuntos
Antioxidantes/farmacologia , Cálcio/metabolismo , Melatonina/farmacologia , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Oócitos/efeitos dos fármacos , Antioxidantes/análise , Feminino , Humanos , Técnicas de Maturação in Vitro de Oócitos , Melatonina/análise , Mitocôndrias/metabolismo , Oócitos/citologia , Oócitos/metabolismo , Estresse Oxidativo
3.
Exp Parasitol ; 204: 107732, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31374184

RESUMO

In the present study, the cytotoxic effects of amitraz, an octopamine receptor agonist on the reproductive system of engorged adult females of Rhipicephalus (Boophilus) annulatus were assessed using histology, electron microscopy and octopamine beta (OCTß) receptor transcriptional expression analysis. Adult immersion test (AIT) was performed by immersing the fully engorged female ticks for 2 min in different concentrations of amitraz (200, 250, 300, 350 ppm). Amitraz at the dose of 300 ppm, caused an adult tick mortality of 16.66 ±â€¯6.80 per cent, inhibition of fecundity of 75.80 per cent and hatching of 50 per cent of ova laid by treated ticks. Histological changes in the ovaries of ticks collected after 24 h of treatment with amitraz (300 ppm), in comparison with controls (distilled water/methanol) were identified by microscopical examination of sections (4  µm) stained using haematoxylin and eosin. These changes included reduction in size and basophilia of stage I oocytes, presence of cytoplasmic vacuoles of various sizes around germinal vesicle of stage II oocytes, wavy basement membrane of stage III oocytes and reduction in size and number of mature stage IV and V oocytes. Electron microscopy was employed for understanding the structural changes in the ultrathin sections (60 nm) of ovaries. Ticks treated with amitraz showed major ultrastructural changes such as irregular nuclear membrane, crystolysis of mitochondria and detachment of external and internal layers of basal lamina of oocytes. The cDNA synthesized from the total RNA of whole ticks and ovaries of ticks treated with amitraz along with controls were used for relative quantification of Octopamine ß receptor (OCTß-R) expression based on the 2-ΔΔCT method by quantitative real time PCR (qRT PCR). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as endogenous control. Down regulation of expression of OCTß-R mRNA in the ovaries of amitraz treated ticks was observed compared to controls. Thus, the inhibition of fecundity observed in the ticks treated with amitraz can be attributed to the major structural changes and decreased expression of OCT ß receptor mRNA induced by it in the ovary.


Assuntos
Inseticidas/farmacologia , Rhipicephalus/efeitos dos fármacos , Toluidinas/farmacologia , Análise de Variância , Animais , Membrana Basal/efeitos dos fármacos , Membrana Basal/ultraestrutura , Regulação para Baixo , Feminino , Fertilidade/efeitos dos fármacos , Expressão Gênica , Microscopia Eletrônica de Transmissão , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/ultraestrutura , Oócitos/efeitos dos fármacos , Oócitos/ultraestrutura , Ovário/anatomia & histologia , Ovário/efeitos dos fármacos , Ovário/ultraestrutura , Oviposição/efeitos dos fármacos , RNA Mensageiro/isolamento & purificação , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Receptores de Amina Biogênica/agonistas , Receptores de Amina Biogênica/efeitos dos fármacos , Rhipicephalus/anatomia & histologia , Rhipicephalus/genética , Rhipicephalus/ultraestrutura , Espectrofotometria , Controle de Ácaros e Carrapatos/métodos , Vacúolos/efeitos dos fármacos , Vacúolos/ultraestrutura
4.
Cell Physiol Biochem ; 53(3): 439-452, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31436397

RESUMO

BACKGROUND/AIMS: Among the assisted reproductive techniques, the in vitro maturation of oocytes (IVM) is less developed than other techniques, but its implementation would entail a qualitative advance. This technique consists in the extraction of immature oocytes from antral ovarian follicles with the patient under low hormone stimulation or without hormone to mature exogenously in culture media supplemented with different molecules to promote maturation. In this sense, we are interested in the role that cannabinoids could have as IVM promoters because cannabinoid's molecular pathway is similar to the one by which oocyte's meiosis resumption is activated. With the intention of advancing in the possible use of cannabinoids as supplements for the media for in vitro maturation of oocytes, we intend to deepen the study of the function of the phytocannabinoid Δ-9-tetrahydrocannabinol (THC) in the IVM process. METHODS: By immunocytochemistry, we detected the location pattern of cannabinoid receptor type 1 (CB1) and type 2 (CB2) during oocyte maturation in presence or absence of THC, as well as, the staining pattern of p-AKT and p-ERK. We used a genetic/ pharmacological approach generating knockout oocytes for CB1 and/or CB2 and they were incubated with THC during the oocyte maturation to visualize the physiological effects of THC, observing the rate of blastocyst achieved by oocyte. RESULTS: This study confirms that the incubation of oocytes with THC during IVM accelerated some events of that process like the phosphorylation pattern of ERK and AKT and was able to increase the blastocyst rate in response to IVF. Moreover, it seems that both CB1 and CB2 are necessary to maintain a healthy oocyte maturation. CONCLUSION: Our data suggest that THC may be useful IVM supplements in clinic as is more feasible and reliable than any synthetic cannabinoid.


Assuntos
Blastocisto/efeitos dos fármacos , Dronabinol/farmacologia , Oócitos/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Blastocisto/citologia , Blastocisto/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Feminino , Fertilização In Vitro , Técnicas de Maturação in Vitro de Oócitos , Meiose/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Oócitos/citologia , Oócitos/metabolismo , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptor CB1 de Canabinoide/genética , Receptor CB1 de Canabinoide/metabolismo , Receptor CB2 de Canabinoide/genética , Receptor CB2 de Canabinoide/metabolismo
5.
Aquat Toxicol ; 215: 105261, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31419757

RESUMO

Harmful cyanobacteria and their production of microcystins (MCs) exert significant toxicity on reproduction of fish, especially the process of oogenesis. Our previous studies demonstrated that MCs have negative impacts on the quantity and quality of mature oocytes in female zebrafish. However, the underlying mechanisms of MCs disrupting oocyte maturation (OM) have been rarely reported. In the present study, in vitro oocytes (immature) were separated from zebrafish and treated with 1, 10, 100 µg/L MC-LR. The serine/threonine protein phosphatase 2A (PP2A) activity was downregulated significantly in oocytes exposed to 10 and 100 µg/L MC-LR for both 2 and 4 h. The phosphorylation levels of mitogen-activated protein kinase (MAPK) were detected without noticeable change in all oocytes treated with MC-LR for 2 h, whereas the activated levels of MAPK subtypes (ERK, p38 and JNK) increased remarkably in the 100 µg/L MC-LR treatment of 4 h. In the oocytes exposed to 100 µg/L MC-LR for 4 h, germinal vesicle breakdown (GVBD) rates changed abnormally and maturation-promoting factor (MPF) activity increased significantly, in accordance with the upregulation of Cyclin B protein levels. Moreover, the MAPK inhibitors (10 µM) were applied to explore the role of MAPK subtypes during MC-LR influencing OM and results showed that ERK inhibitor U0126 and p38 inhibitor SB203580 mitigated the effects of 100 µg/L MC-LR-induced MAPK hyper-phosphorylation and elevated GVBD in the oocytes. In conclusion, the present study indicates that microcystins disrupt the meiotic maturation by the pathway of MC-PP2A-MAPK-OM due to the phosphorylation disorder in oocytes.


Assuntos
Técnicas de Maturação in Vitro de Oócitos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Microcistinas/toxicidade , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Peixe-Zebra/fisiologia , Animais , Ciclina B/metabolismo , Ativação Enzimática/efeitos dos fármacos , Feminino , Fator Promotor de Maturação/metabolismo , Oócitos/efeitos dos fármacos , Oócitos/enzimologia , Fosforilação/efeitos dos fármacos , Inibidores de Proteínas Quinases/farmacologia , Proteína Fosfatase 2/metabolismo , Vitelogeninas/metabolismo , Poluentes Químicos da Água/toxicidade
6.
Life Sci ; 232: 116639, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31295472

RESUMO

AIMS: Sirtuins have been implicated in the aging process, however, the functions of SIRT2 in post-maturation aging of oocytes are not fully understood. The purpose of the present investigation was to assess the roles of SIRT2 in aged oocytes and mechanisms involved. MAIN METHODS: The fresh MII oocytes were aging in vitro, and treated with SIRT2 inhibitor (SirReal2), autophagy activator (Rapamycin), and autophagy inhibitor (3-Ma) for 24 h, respectively. Oocyte activation, cytoplasmic fragmentation, and spindle defects, mitochondrial distribution, ROS levels, ATP production, mitochondrial membrane potential, and early apoptosis were investigated. Western blotting was performed to determine LC3-II accumulation, SQSTM1 degradation, and caspase-3 activity. KEY FINDINGS: SIRT2 expression gradually decreased in a time-dependent manner during oocyte aging. Treatment with SirReal2 significantly increased the rates of oocyte activation, cytoplasmic fragmentation, and spindle defects. In particular, the high ROS levels, abnormal mitochondrial distribution, low ATP production, and lost ΔΨm were observed in SirReal2-exposed oocytes. Further analysis revealed that LC3-II accumulation and SQSTM1 degradation were induced by SIRT2 inhibition. By performing early apoptosis analysis showed that oocyte aging was accompanied with cellular apoptosis, and SIRT2 inhibition increased apoptosis rates of aged oocytes. Importantly, upregulating autophagy with Rapamycin could mimic the effects of SIRT2 inhibition on apoptosis by increasing caspase-3 activation, whereas downregulating autophagy with 3-MA could abolish those effects by blocking caspase-3 activation. SIGNIFICANCE: Our results suggest that SIRT2 inactivation is a key mechanism underlying of cellular aging, and SIRT2 inhibition contributes to autophagy-dependent cellular apoptosis in post-maturation oocytes.


Assuntos
Oócitos/fisiologia , Sirtuína 2/fisiologia , Acetamidas/farmacologia , Animais , Apoptose/efeitos dos fármacos , Apoptose/fisiologia , Autofagia/efeitos dos fármacos , Autofagia/fisiologia , Bovinos , Senescência Celular/efeitos dos fármacos , Senescência Celular/fisiologia , Feminino , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Oócitos/classificação , Oócitos/efeitos dos fármacos , Oócitos/metabolismo , Sirolimo/farmacologia , Sirtuína 2/antagonistas & inibidores , Sirtuína 2/metabolismo , Tiazóis/farmacologia
7.
Environ Pollut ; 252(Pt A): 388-398, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31158667

RESUMO

Dibutyl phthalate (DBP), one of the most widely used plasticizers, is a known environmental endocrine disruptor that impairs male and female fertility. In this study, oral administration of DBP was given to pregnant mice on 14.5 days post coitus (dpc) for 3 days; and additionally, DBP was added into the culture of 14.5 dpc fetal ovaries for 3 days. DBP exposure during gestation disturbed the progression of meiotic prophase I of mouse oocytes, specifically from the zygotene to pachytene stages. Meanwhile, the DBP-exposed pachytene oocytes showed increased homologous recombination sites and unrepaired DNA damage. Furthermore, DBP caused DNA damage by increasing oxidative stress, decreased the expression of multiple critical meiotic regulators, and consequently induced oocyte apoptosis. Moreover, the effect of DBP on meiosis I prophase involved estrogen receptors α and ß. Collectively, these results demonstrated a set of meiotic defects in DBP-exposed fetal oocytes. As aberrations in homologous recombination can result in aneuploid gametes and embryos, this study provides new support for the deleterious effects of phthalates.


Assuntos
Dibutilftalato/toxicidade , Disruptores Endócrinos/toxicidade , Recombinação Homóloga/efeitos dos fármacos , Prófase Meiótica I/efeitos dos fármacos , Oócitos/efeitos dos fármacos , Plastificantes/toxicidade , Aneuploidia , Animais , Apoptose/efeitos dos fármacos , Feminino , Masculino , Prófase Meiótica I/genética , Camundongos , Oócitos/patologia , Ovário/efeitos dos fármacos , Ovário/patologia , Gravidez
8.
Ecotoxicol Environ Saf ; 181: 370-380, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31212185

RESUMO

Cigarette smoke can cause follicle destruction and oocyte dysfunction and increase the risks of spontaneous abortion, stillbirth, and tubal ectopic pregnancy, affecting female reproductive health. Third-hand smoke (THS) is residual tobacco smoke existing in the environment long after cigarettes are extinguished, which can react with other compounds in the environment to produce secondary pollutants. However, the effects of THS on the female reproductive system, particularly the maturation of the oocyte, remain unclear. 1-(N-methyl-N-nitrosamino)-1-(3-pyridinyl)-4-butanal (NNA), a component of THS, is a logical biomarker of THS exposure. Thus, this study aims to investigate the toxic effects of NNA on the maturation of murine oocytes and subsequent developmental competence. Herein, murine oocytes were exposed to 0 (control group), 0.1, 1.0, 10, and 50 µM NNA for 24 h. Our results showed that NNA exposure reduced the polar body extrusion rate by causing 8-oxo-deoxyguanosine (8-OHdG) to increase and disrupting the meiotic spindle morphology by inhibiting ERK1/2 activation during in vitro maturation. Additionally, NNA exposure resulted in cleavage and blastocyst rate reduction by altering DNA and histone methylations by reducing 5 mC and H3K4me2 levels and by inducing apoptosis caused by mitochondrial dysfunction and reactive oxygen species accumulation, as shown by the increased superoxide dismutase mRNA level and by the decreased Bcl-x mRNA level. Collectively, our results demonstrate that NNA exposure reduces the maturation and developmental capability of murine oocytes by increasing the risk of DNA damage and abnormal spindle morphology, altering epigenetic modifications, and inducing apoptosis, suggesting the toxic effect of NNA on mammalian productive health.


Assuntos
Aldeídos/toxicidade , Poluentes Ambientais/toxicidade , Nitrosaminas/toxicidade , Oócitos/efeitos dos fármacos , Piridinas/toxicidade , Animais , Apoptose , Dano ao DNA , Epigênese Genética , Feminino , Camundongos , Oócitos/crescimento & desenvolvimento , Oócitos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Fuso Acromático/efeitos dos fármacos
9.
Ceska Gynekol ; 84(2): 161-165, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31238688

RESUMO

OBJECTIVE: Bisphenols are one of the most widespread endocrine disruptors that the population of west world countries is exposed to. Objective of this study is to summarize information about influence of bisphenols on reproduction health. DESIGN: Review article, Setting: Department of Obstetrics and Gynecology, Faculty of Medicine, Masaryk University and University Hospital Brno. METHODS: PubMed was searched for articles in English indexed bisphenol and reproduction up to October 2018. RESULTS: Increased levels of bisphenol A and S have been proven in body fluids and tissues. Bisphenol molecules have effect similar to estrogens therefore they influence hormonal regulation and activity of estrogen receptors. Their negative influence on oocyte maturation, spermatogenesis and development of reproductive system has been shown. Bisphenol S, which has replaced bisphenol A, has comparable negative effects on reproduction. CONCLUSION: Bisphenols are widespread endocrine disruptors that could cause severe fertility disorders of men and women.


Assuntos
Compostos Benzidrílicos/farmacologia , Disruptores Endócrinos/farmacologia , Estrogênios não Esteroides/farmacologia , Oócitos/efeitos dos fármacos , Fenóis/farmacologia , Reprodução/efeitos dos fármacos , Espermatogênese/efeitos dos fármacos , Feminino , Humanos , Masculino , Gravidez
10.
Environ Toxicol ; 34(9): 1001-1012, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31112002

RESUMO

Ginsenoside Rb1 (GRb1), the major saponin component of ginseng root, has a wide range of therapeutic applications for various diseases. Previously, our group showed that GRb1 triggers ROS-mediated apoptotic cascades in mouse blastocysts, leading to decreased cell viability and impairment of pre- and postimplantation embryonic development, both in vitro and in vivo. In this study, we further found that GRb1 exerted dose-dependent effects on oocyte maturation and sequent development in vitro. Oocytes preincubated with 25 µg/mL GRB1 displayed significantly enhanced maturation and in vitro fertilization (IVF) rates, along with progression of subsequent embryonic development. In contrast, treatment with 50 and 100 µg/mL GRB1 led to impairment of mouse oocyte maturation, decreased IVF rates, and injurious effects on subsequent embryonic development. In vivo, intravenous injection of 1 mg/kg body weight GRb1 significantly promoted mouse oocyte maturation, IVF, and early-stage embryo development after fertilization while administration of 5 mg/kg body weight GRb1 led to a marked decrease in oocyte maturation and IVF rates concomitant with impairment of early embryonic development in our animal model. In terms of the mechanisms underlying the regulatory effects of GRb1 demonstrated increased intracellular reactive oxygen species (ROS) production and apoptosis in the 100 µg/mL GRb1 treatment group. However, we observed a significant decrease in total intracellular ROS content and inhibition of apoptosis events in the 25 µg/mL GRb1 treatment group, signifying that the intracellular ROS content serves as a key upstream regulator of GRb1 that influences its dose-dependent beneficial or deleterious effects on oocyte maturation and sequent embryonic development. For further clarification of the mechanisms underlying GRb1-triggered injurious effects, oocytes were pretreated with Ac-DEVD-CHO, a caspase-3-specific inhibitor, which effectively blocked injury to oocyte maturation, fertilization, and sequent development. In sum, study findings highlight the potential involvement of p53-, p21-, and caspase-3-dependent regulatory signaling cascades in GRb1-mediated apoptotic processes.


Assuntos
Desenvolvimento Fetal/efeitos dos fármacos , Ginsenosídeos , Oócitos/efeitos dos fármacos , Oogênese/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Blastocisto/efeitos dos fármacos , Blastocisto/metabolismo , Relação Dose-Resposta a Droga , Desenvolvimento Embrionário/efeitos dos fármacos , Feminino , Fertilização In Vitro , Ginsenosídeos/administração & dosagem , Ginsenosídeos/farmacologia , Ginsenosídeos/toxicidade , Técnicas In Vitro , Camundongos , Camundongos Endogâmicos ICR , Oócitos/metabolismo , Gravidez , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais
11.
Mol Med Rep ; 19(6): 5353-5360, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31059097

RESUMO

Ppm1b, a metal­dependent serine/threonine protein phosphatase, catalyzes the dephosphorylation of a variety of phosphorylated proteins. Ppm1b­/­ mouse embryos die at the fertilized oocyte stage, whereas Ppm1b+/­ mice with a C57BL/6 background exhibit no phenotypic abnormalities. Because the C57BL/6 strain produces a limited number of pups, in an attempt to produce Ppm1b­/­ mice, congenic Ppm1b+/­ mice with an ICR background were established, which are more fertile and gave birth to more pups. As a result, however, no Ppm1b­/­ offspring were obtained when pairs of Ppm1b+/­ ICR mice were bred again. Ppm1b+/­ male and female ICR mice were analyzed from the viewpoint of fecundity. The Ppm1b haploinsufficiency had no effect on testicular weight or the number of sperm in male mice. Despite the fact that the levels of Ppm1b protein in the ovaries of sexually mature Ppm1b+/­ mice were decreased compared with those of Ppm1b+/+ mice, there appeared to be no significant difference in the histological appearance of the ovaries, litter sizes or plasma progesterone levels at the estrous stage. When superovulation was induced by stimulation using a hormone treatment, the number of ovulated oocytes were the same for Ppm1b+/­ and Ppm1b+/+ mice at 4 weeks of age when the estrous cycle did not proceed, however, the number of ovulated oocytes was lower in sexually mature Ppm1b+/­ mice at 11 weeks of age compared with Ppm1b+/+ mice in the first and the second superovulation cycles. These collective results suggest that follicle development is excessive in Ppm1b+/­ mice, and that this leads to a partial depletion of matured follicles and a corresponding decrease in the number of ovulated oocytes.


Assuntos
Proteína Fosfatase 2C/genética , Superovulação , Animais , Gonadotropina Coriônica/farmacologia , Feminino , Heterozigoto , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos ICR , Oócitos/citologia , Oócitos/efeitos dos fármacos , Oócitos/metabolismo , Ovário/metabolismo , Gravidez , Progesterona/sangue , Proteína Fosfatase 2C/metabolismo , Superovulação/efeitos dos fármacos
12.
Anim Reprod Sci ; 205: 94-104, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31060922

RESUMO

The effect of heat stress (HS) on cattle reproduction is deleterious with respect to ovarian follicular development and oocyte quality. The objective of this study was to investigate the effect of follicular fluid extracellular vesicles (EVs) obtained from cows maintained in thermoneutral (TN) or HS conditions on in vitro oocyte maturation. Nonlactating cows were estrous synchronized. Immediately after ovulation day (D1), the cows were randomly assigned to TN or HS environments. Follicular fluid from all follicles from each treatment was pooled, and EVs were obtained. Pools of 20 cumulus oocyte-complexes (COCs), were allocated to the following treatments: Control (n = 4 COC pools): matured in base medium; TN (n = 4 COC pools): matured in base medium supplemented with TN EV suspension; and HS (n = 4 COC pools): matured in base medium that was supplemented with the HS EV suspension. All treatments were conducted at 38.5 °C for 24 h in a humid atmosphere with 5% CO2. After maturation, the COCs were evaluated for meiotic progression, DNA integrity and oocyte quality-related gene expression. When the experimental groups were compared with the control group, a treatment effect was not observed for meiotic progression and DNA integrity. In the cumulus cells of TN group, there was relatively lesser expression of the IGFBP4 gene. In the oocytes of the TN as compared with the HS group, the IGFBP2, BMP15, GDF9, CDCA8, HAS2, RPL15, STAT3 and PFKP genes were expressed to a lesser extent. The findings indicated that oocytes matured in the presence of EVs from the follicular fluid of cows collected when there were TN conditions, however, there was a lesser expression of genes related to oocyte quality.


Assuntos
Doenças dos Bovinos/metabolismo , Fertilização In Vitro/veterinária , Líquido Folicular , Transtornos de Estresse por Calor/veterinária , Técnicas de Maturação in Vitro de Oócitos/veterinária , Oócitos/efeitos dos fármacos , Animais , Bovinos , Células do Cúmulo , Feminino , Temperatura Alta , Folículo Ovariano
13.
Anim Reprod Sci ; 205: 150-155, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31076217

RESUMO

As oocytes and embryos of pigs have greater lipid content in the cytoplasm than those of other species, supplementation of the medium for in vitro maturation (IVM) of oocytes with omega-3 polyunsaturated fatty acids (PUFA) may help to improve embryo development. This study was conducted to evaluate effects of the inclusion of the docosaexaenoic (DHA) and of the eicosapentaenoic acids (EPA) in the IVM medium on the development of pig oocytes and on the lipid content of oocytes and embryos. In all experiments, control media consisted of porcine follicular fluid and oocytes were activated through parthenogenesis. In Experiment 1, there were four treatments for each PUFA: one control; and three treatments including EPA or DHA in the IVM medium at 12.5 µM, 25.0 µM and 50.0 µM). In Experiment 2, inclusion of 50 µM DHA was compared against the control. Cleavage rates in the IVM medium including 12.5 µM EPA and blastocyst development rates in media at any EPA concentration were less than for the control in Experiment 1 (P < 0.05). Compared to the control, inclusion of 50 µM DHA in the IVM medium was related to greater cleavage rates and greater number of embryo cells, in Experiment 1, and lesser lipid content in oocytes after 22 and 44 h and in embryos after 7 days, in Experiment 2 (both P < 0.05). Addition of DHA in the IVM medium may benefit the development of pig oocytes, but EPA appears to be cytotoxic.


Assuntos
Ácidos Docosa-Hexaenoicos/farmacologia , Ácido Eicosapentaenoico/farmacologia , Embrião de Mamíferos/química , Técnicas de Maturação in Vitro de Oócitos/veterinária , Oócitos/efeitos dos fármacos , Suínos/embriologia , Animais , Ácidos Docosa-Hexaenoicos/administração & dosagem , Relação Dose-Resposta a Droga , Ácido Eicosapentaenoico/administração & dosagem , Feminino , Metabolismo dos Lipídeos , Partenogênese , Suínos/fisiologia
14.
Environ Int ; 128: 390-398, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31078873

RESUMO

Dydrogesterone (DDG) is a synthetic progestin widely used in numerous gynecological diseases. DDG has been shown to disturb fish reproduction, however, the mechanism is still unclear. Here we studied the histological changes and differences of metabolome between exposed and control fish gonads after exposure of zebrafish (Danio rerio) embryos to 2.8, 27.6, and 289.8 ng/L DDG until sexual maturity for a total of 140 days. Dydrogesterone exposure led to male-biased zebrafish sex ratios. Histological examination revealed that DDG induced postovulatory follicles and atretic follicles in the ovary of the female fish. Postovulatory follicles indicated the occurrence of ovulation. DDG also increased spermatids and spermatozoa in the male fish testis, suggesting promotion of spermatogenesis. Ovarian metabolome showed that DDG increased the concentrations of free amino acids, urea, putrescine, free fatty acids, acylcarnitines, lysophospholipids, and other metabolites catabolized from phospholipids. Most of these metabolites are biodegradation products of proteins and lipids, suggesting the existence of ovulated oocytes over-ripening. Further, DDG upregulated arachidonic acid (AA) and its 5­lipoxygenase (5-LOX) metabolites 5­oxo­6,8,11,14­eicosatetraenoic acid (5-oxo-ETE) in the ovary, which could lead to suppression of AA cyclooxygenase (COX) metabolite prostaglandin F2α (PGF2α). It is believed that AA induced oocyte maturation, while 5-oxo-ETE and related metabolites in purinergic signaling promoted ovulation. Whereas, the suppression of PGF2α production might block spawning and damaged follicular tissue digestion, which explained the oocytes over-ripening and atretic follicles in the treated ovary. Overall, our results suggested that DDG exposure induced zebrafish oocyte maturation and ovulation but led to oocytes over-ripening via the AA metabolic pathway and purinergic signaling.


Assuntos
Didrogesterona/efeitos adversos , Disruptores Endócrinos/efeitos adversos , Oócitos/efeitos dos fármacos , Ovulação/efeitos dos fármacos , Poluentes Químicos da Água/efeitos adversos , Peixe-Zebra/fisiologia , Animais , Relação Dose-Resposta a Droga , Embrião não Mamífero/efeitos dos fármacos , Metaboloma/efeitos dos fármacos , Metabolômica , Progestinas/efeitos adversos , Distribuição Aleatória , Reprodução/efeitos dos fármacos , Razão de Masculinidade
15.
Int J Mol Sci ; 20(11)2019 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-31142052

RESUMO

Somatic cell nuclear transfer (SCNT) is a very powerful technique used to produce genetically identical or modified animals. However, the cloning efficiency in mammals remains low. In this study, we aimed to explore the effects of vitamin C (Vc)-treated donor cells on cloned embryos. As a result, Vc treatment relaxed the chromatin of donor cells and improved cloned embryo development. RNA sequencing was adopted to investigate the changes in the transcriptional profiles in early embryos. We found that Vc treatment increased the expression of genes involved in the cell-substrate adherens junction. Gene ontology (GO) analysis revealed that Vc treatment facilitated the activation of autophagy, which was deficient in cloned two-cell embryos. Rapamycin, an effective autophagy activator, increased the formation of cloned blastocysts (36.0% vs. 25.6%, p < 0.05). Abnormal expression of some coding genes and long non-coding RNAs in cloned embryos was restored by Vc treatment, including the zinc-finger protein 641 (ZNF641). ZNF641 compensation by means of mRNA microinjection improved the developmental potential of cloned embryos. Moreover, Vc treatment rescued some deficient RNA-editing sites in cloned two-cell embryos. Collectively, Vc-treated donor cells improved the development of the cloned embryo by affecting embryonic transcription. This study provided useful resources for future work to promote the reprogramming process in SCNT embryos.


Assuntos
Ácido Ascórbico/farmacologia , Blastocisto/efeitos dos fármacos , Clonagem de Organismos/métodos , Oócitos/efeitos dos fármacos , Transcriptoma , Vitaminas/farmacologia , Animais , Autofagia , Blastocisto/metabolismo , Bovinos/genética , Células Cultivadas , Feminino , Masculino , Técnicas de Transferência Nuclear , Oócitos/metabolismo
16.
BMC Plant Biol ; 19(1): 190, 2019 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-31068146

RESUMO

BACKGROUND: The functional characteristics of SLAC/SLAH family members isolated from Arabidopsis thaliana, poplar, barley and rice have been comprehensively investigated. However, there are no reports regarding SLAC/SLAH family genes from Rosaceae plants. RESULTS: In this study, the function of PbrSLAH3, which is predominately expressed in pear (Pyrus bretschneideri) root, was investigated. PbrSLAH3 can rescue the ammonium toxicity phenomenon of slah3 mutant plants under high-ammonium/low-nitrate conditions. In addition, yeast two-hybrid and bimolecular fluorescence complementation assays confirmed that PbrSLAH3 interacts with PbrCPK32. Moreover, when PbrSLAH3 was co-expressed with either the Arabidopsis calcium-dependent protein kinase (CPK) 21 or PbrCPK32 in Xenopus oocytes, yellow fluorescence was emitted from the oocytes and typical anion currents were recorded in the presence of extracellular NO3-. However, when PbrSLAH3 alone was injected, no yellow fluorescence or anion currents were recorded, suggesting that anion channel PbrSLAH3 activity was controlled through phosphorylation. Finally, electrophysiological and transgene results showed that PbrSLAH3 was more permeable to NO3- than Cl-. CONCLUSION: We suggest that PbrSLAH3 crossing-talk with PbrCPK32 probably participate in transporting of nitrate nutrition in pear root.


Assuntos
Canais Iônicos/metabolismo , Nitratos/metabolismo , Proteínas de Plantas/metabolismo , Proteínas Quinases/metabolismo , Pyrus/enzimologia , Compostos de Amônio/toxicidade , Animais , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Fenômenos Eletrofisiológicos , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes de Plantas , Mutação/genética , Oócitos/efeitos dos fármacos , Oócitos/metabolismo , Filogenia , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Pyrus/efeitos dos fármacos , Pyrus/genética , Xenopus
17.
Wei Sheng Yan Jiu ; 48(1): 109-113, 2019 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-31032778

RESUMO

OBJECTIVE: To study the effects of cigarette smoke on the oxidative damage of mice oocytes and developmental potential. METHODS: A total of 96 SPF grade 6-week-old ICR female mice were randomly divided into control group, low-dose group, middle-dose group and high-dose group according to the principle of similar body weight. Twenty-four mice in each group were placed in a cigarette-lighting containers with the volume of 60, 32 and 23 L respectively. The control group was fed normally twice a day for 1 hour for 4 weeks. Weight and record daily, draw weight change curve; use kit to detect the activity of superoxide dismutase(SOD) and glutathione peroxidase(GPX) in ovarian tissue; use tissue section, indirect immunofluorescence and in vitro maturation technology to detect the activity of each group. The ovarian organ index, the level of reactive oxygen species(ROS) in oocyte and the first polar body rate were measured. The t test was used to compare the mean values of the two groups, and one-way ANOVA was used to compare the mean values of the multiple groups. RESULTS: Compared with the control group, the weight gain of mice in the treatment group was significantly slower(the weight gain in the high dose group was only 1. 05%±0. 41%). The ovarian organ index, SOD and GPX activity in the ovarian tissue and litter size(the high dose group was reduced to 5. 20%±1. 48%) were significantly decreased. The ROS level in oocyte was significantly increased, and oocyte maturation in vitro was the first. The polar body rate decreased(high dose group decreased to 49. 34%±5. 78%), and the difference was statistically significant(P<0. 05). CONCLUSION: Cigarette smoke exposure in mice can cause the decrease of ovarian organ index, SOD and GPX activity, the increase of ROS content in oocyte, the decrease of first polar body rate and litter size.


Assuntos
Oócitos , Estresse Oxidativo , Fumar , Animais , Feminino , Camundongos , Camundongos Endogâmicos ICR , Oócitos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio , Fumaça/efeitos adversos , Fumar/efeitos adversos , Superóxido Dismutase
18.
Anim Reprod Sci ; 205: 62-69, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31005360

RESUMO

The aims of the present study were to determine the concentrations of lipopolysaccharide (LPS), hormonal progesterone, estradiol-17ß, insulin growth factor (IGF-1) and magnesium in the serum and the preovulatory follicle follicular fluid (FF) in repeat breeder (RB) cows without (nSCE) or with subclinical endometritis (SCE), and further to examine the effects of this FF on developmental competence of cattle oocytes. In Experiment 1, 13 of 23 clinically healthy Holstein RB cows were identified (uterine PMNs) to have SCE. The cows were estrous synchronized, and 6-12 h after detection of standing estrus, FF and blood of the preovulatory follicles were collected. The mean (±SD) LPS (862.3 ± 148.1 compared with 1063.4 ± 262.8 EU/ml, P = 0.04) and estradiol-17ß (188.9 ± 15.8 compared with 162.0 ± 31.5 ng/ml, P = 0.02) concentrations of FF was different between nSCE and SCE cows. In Experiment 2, FF of RB cows with relatively lesser (nSCE, n = 4) and greater (SCE, n = 4) percentages of uterine PMNs was separately added to the oocyte maturation medium for in vitro embryo production. Addition of FF from SCE cows to the oocyte maturation medium resulted in a lesser rate of development to the blastocyst stage than that of the nSCE cows (21.9 ± 1.8 compared with 27.8 ± 2.5%, P < 0.05). Results of the present study indicate greater FF LPS concentration may result in a lesser quality microenvironment milieu for the final stages of oocyte maturation in RB dairy cows with subclinical endometritis. In addition, supplementation of oocyte maturation medium with FF of preovulatory follicles from RB cows with subclinical endometritis resulted in a lesser potential of in vitro oocyte developmental competence.


Assuntos
Doenças dos Bovinos/metabolismo , Endometrite/veterinária , Líquido Folicular , Oócitos/efeitos dos fármacos , Animais , Bovinos , Endometrite/metabolismo , Sincronização do Estro , Feminino , Masculino , Oócitos/fisiologia , Folículo Ovariano , Ovulação , Capacitação Espermática/efeitos dos fármacos
19.
Zygote ; 27(2): 55-63, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30871647

RESUMO

SummaryStudies have shown that daily exposure to different products, whether chemical or natural, can cause irreversible damage to women's reproductive health. Therefore it is necessary to use tests that evaluate the safety and efficacy of these products. Most reproductive toxicology tests are performed in vivo. However, in recent years, various cell culture methods, including embryonic stem cells and tissues have been developed with the aim of reducing the use of animals in toxicological tests. This is a major advance in the area of toxicology, as these systems have the potential to become a widely used tool compared with in vivo tests routinely used in reproductive biology and toxicology. The present review describes and highlights data on in vitro culture processes used to evaluate reproductive toxicity as an alternative to traditional methods using in vivo tests.


Assuntos
Técnicas de Cultura de Órgãos/métodos , Folículo Ovariano/citologia , Ovário/efeitos dos fármacos , Reprodução/efeitos dos fármacos , Testes de Toxicidade/métodos , Animais , Linhagem Celular , Feminino , Humanos , Oócitos/citologia , Oócitos/efeitos dos fármacos , Folículo Ovariano/efeitos dos fármacos , Ovário/fisiologia
20.
Ticks Tick Borne Dis ; 10(4): 766-774, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30904538

RESUMO

Rhipicephalus microplus, commonly known as southern cattle tick, causes huge economic losses in the cattle industry. Its infestation affects the production of meat and milk and causes discomfort to hosts. In addition, it is the vector of Babesia spp. and Anaplasma spp. The most frequent method used to control these parasites is through synthetic acaricides. However, their indiscriminate use can be toxic for hosts and environment as well as cause selection of resistant ticks. Plant extracts and essential oils emerge as promising alternatives to manage tick infestation. Carvacrol, an aromatic monoterpene extracted from plants, has recognized antimicrobial, antioxidant, insecticidal, repellent and acaricidal activities. Acetylation of carvacrol is believed to enhance its nematicidal and acaricidal activities and to decrease its toxicity to hosts. Thus, the aim of this study was to evaluate the effect of different concentrations of acetylcarvacrol in the morphology of ovaries of engorged R. microplus ticks. The most remarkable morphological alterations found in the female germ cells were irregular and thicker chorion, decreasing in size and irregular shape of female germ cells (oocytes), cytoplasmatic vacuolization as well as ring-shaped nucleoli. These alterations were analyzed through a semi-quantitative method proposed in this study for ixodid ticks. Treatment group IV, which was exposed to 4.5 µL/mL of carvacrol acetate, showed the most significant alterations, and it was also statistically different when compared to control groups. Therefore, sublethal concentrations of acetylcarvacrol demonstrated to impact the reproductive system of R. microplus by causing several damages in the female germ cells. This would hinder the generation of new individuals, probably contributing for a long-term control of tick infestation.


Assuntos
Acaricidas , Monoterpenos/química , Oócitos/efeitos dos fármacos , Óleos Vegetais/química , Rhipicephalus , Infestações por Carrapato/veterinária , Animais , Bovinos , Feminino , Óleos Voláteis/química , Oócitos/crescimento & desenvolvimento , Oogênese/efeitos dos fármacos , Controle de Ácaros e Carrapatos/métodos , Infestações por Carrapato/tratamento farmacológico
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