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1.
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
2.
Yonsei Med J ; 60(7): 667-678, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31250581

RESUMO

PURPOSE: The aim of this study was to investigate how type I diabetes mellitus (T1D) affects the folliculogenesis and oocyte development, fertilization, and embryo development. MATERIALS AND METHODS: A comparative animal study was conducted using two different mouse models of T1D, a genetic AKITA model and a streptozotocin-induced diabetes model. Ovarian function was assessed by gross observation, immunoblot, immunohistochemistry, oocyte counting, and ELISA for serum hormones (insulin, anti-Mullerian hormone, estradiol, testosterone, and progesterone). Maturation and developmental competence of metaphase II oocytes from control and T1D animals was evaluated by immunofluorescent and immunohistochemical detection of biomarkers and in vitro fertilization. RESULTS: Animals from both T1D models showed increased blood glucose levels, while only streptozotocin (STZ)-injected mice showed reduced body weight. Folliculogenesis, oogenesis, and preimplantation embryogenesis were impaired in both T1D mouse models. Interestingly, exogenous streptozotocin injection to induce T1D led to marked decreases in ovary size, expression of luteinizing hormone/chorionic gonadotropin receptor in the ovaries, the number of corpora lutea per ovary, oocyte maturation, and serum progesterone levels. Both T1D models exhibited significantly reduced pre-implantation embryo quality compared with controls. There was no significant difference in embryo quality between STZ-injected and AKITA diabetic mice. CONCLUSION: These results suggest that T1D affects folliculogenesis, oogenesis, and embryo development in mice. However, the physiological mechanisms underlying the observed reproductive effects of diabetes need to be further investigated.


Assuntos
Diabetes Mellitus Tipo 1/patologia , Fertilidade , Hormônio Luteinizante/metabolismo , Oócitos/patologia , Ovário/patologia , Receptores do LH/metabolismo , Transdução de Sinais , Animais , Glicemia/metabolismo , Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Experimental/patologia , Diabetes Mellitus Experimental/fisiopatologia , Diabetes Mellitus Tipo 1/sangue , Diabetes Mellitus Tipo 1/fisiopatologia , Feminino , Humanos , Camundongos , Camundongos Endogâmicos C57BL
3.
Maturitas ; 125: 1-4, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31133209

RESUMO

Egg freezing was initially used as an intervention for iatrogenic ovarian insufficiency. Nowadays, it is not considered an experimental procedure, and it is increasingly popular among healthy women who wish to maintain their reproductive potential (fertility preservation). This review summarises the evidence on egg freezing technology, reproductive outcomes and long-term effects, as well as its psychological and socioeconomic implications. Egg freezing technology is advancing, with vitrification being the current method of choice, due to its effectiveness. The reproductive outcomes following egg freezing appear to be promising, though the strong negative association with maternal age has to be taken into consideration. Unfortunately, data on long-term outcomes are missing, raising safety issues. Egg freezing has several socioeconomic implications involving health risks, as well as financial and psychological factors.


Assuntos
Criopreservação/métodos , Preservação da Fertilidade/métodos , Idade Materna , Oócitos/patologia , Adulto , Tomada de Decisões , Medicina Baseada em Evidências , Feminino , Humanos , Pessoa de Meia-Idade , Reserva Ovariana , Reprodução , Risco , Classe Social , Resultado do Tratamento , Vitrificação
4.
Ecotoxicol Environ Saf ; 180: 168-178, 2019 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-31082581

RESUMO

Fluorene-9-bisphenol (BHPF), a substitute for bisphenol A, is a chemical component of plastics for industrial production. There is evidence that BHPF exerts an antioestrogenic effect on mice, induces endometrial atrophy and leads to adverse pregnancy outcomes. However, the effects of BHPF on oocyte maturation and ovary development as well as its possible mechanisms remain unclear. The objective of this study was to investigate the toxicity and mechanism of BHPF exposure in mouse oocytes in vitro and in vivo. Our results showed that BHPF could inhibit the maturation of oocytes in vitro by reducing the protein level of p-MAPK and destroying the meiotic spindle. We found that in vitro, BHPF-treated oocytes showed increased ROS levels, DNA damage, mitochondrial dysfunction, and expression of apoptosis- and autophagy-related genes, such as Bax, cleaved-caspase 3, LC 3 and Atg 12. In addition, in vivo experiments showed that BHPF exposure could induce the expression of oxidative stress genes (Cat, Gpx 3 and Sod 2) and apoptosis genes (Bax, Bcl-2 and Cleaved-caspase 3) and increase the number of atresia follicles in the ovaries. Our data showed that BHPF exposure affected the first polar body extrusion of oocytes, increased oxidative stress, destroyed spindle assembly, caused DNA damage, altered mitochondrial membrane potentials, induced apoptosis and autophagy, and affected ovarian development.


Assuntos
Compostos Benzidrílicos/toxicidade , Disruptores Endócrinos/toxicidade , Fluorenos/toxicidade , Oócitos/patologia , Ovário/patologia , Fenóis/toxicidade , Animais , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Dano ao DNA/efeitos dos fármacos , Feminino , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos , Oócitos/crescimento & desenvolvimento , Oócitos/metabolismo , Oogênese/efeitos dos fármacos , Ovário/crescimento & desenvolvimento , Ovário/metabolismo , Estresse Oxidativo/efeitos dos fármacos
5.
J Hum Genet ; 64(5): 379-385, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30765866

RESUMO

Oocyte maturation arrest results in primary female infertility, but the genetic etiology of this phenotype remains largely unknown. Previously, we and other groups have reported that biallelic mutations in PATL2 are mainly responsible for human oocyte germinal vesicle-stage arrest and that the specific phenotype varies for different mutations. Here, we identified four novel missense mutations (p.V260M, p.Q300*, p.T425P, and p.D293Y), a novel frameshift mutation (p.N239Tfs*9), and a reported splicing mutation (p.R75Vfs*21) in PATL2 in seven affected individuals from five unrelated families, showing a multiplicity of phenotypes in oocyte maturation arrest, fertilization failure, or embryonic developmental arrest, which further expands the mutational and phenotypic spectrum in patients with PALTL2 mutations. This work further indicates the critical role of PATL2 in oocyte maturation and early embryo development and will provide a basis for pursuing the determination of genetic variation in PALT2 as an additional criterion for evaluating the quality of oocytes and embryos for assisted reproduction techniques.


Assuntos
Infertilidade Feminina/genética , Mutação de Sentido Incorreto , Proteínas Nucleares/genética , Proteínas de Ligação a RNA/genética , Substituição de Aminoácidos , Embrião de Mamíferos/embriologia , Embrião de Mamíferos/patologia , Desenvolvimento Embrionário/genética , Feminino , Humanos , Infertilidade Feminina/metabolismo , Infertilidade Feminina/patologia , Proteínas Nucleares/metabolismo , Oócitos/metabolismo , Oócitos/patologia , Proteínas de Ligação a RNA/metabolismo
6.
Mar Pollut Bull ; 140: 341-352, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30803653

RESUMO

We studied the bioaccumulation of metals and PAHs, the pathological conditions, regressive phenomena and pathogens in wild Mytilus galloprovincialis taken along the North Pier facing the former second Italian largest steelworks of Bagnoli. There was no Cd and Pb bioaccumulation with respect to the EU role 221/2002. Metal shell index decreased as follows: Pb > Ni > Zn > Cu and correlates with the pollution state. The level of BaP was up to thirtysixfold higher the EU rule 835/2011. The sum of 4 hydrocarbons, PAH4, were up to seventeen-fold the rule. PAH levels increased toward the coast. Prevalence values of tissue necrosis and inflammatory lesions were between 50 and 100%. In May animals showed lesion like granulocytomas and inflammatory capsules. Signs of atresia, necrotic oocytes and diffused cases of hermaphroditism were detected. An appropriate localization of farming to avoid contamination from sediment turbulence and risks for consumer health is needed.


Assuntos
Monitoramento Ambiental/métodos , Metais Pesados/análise , Mytilus/efeitos dos fármacos , Hidrocarbonetos Policíclicos Aromáticos/metabolismo , Poluentes Químicos da Água/metabolismo , Animais , Sedimentos Geológicos/química , Itália , Mar Mediterrâneo , Metais Pesados/metabolismo , Mytilus/metabolismo , Necrose , Oócitos/efeitos dos fármacos , Oócitos/patologia , Hidrocarbonetos Policíclicos Aromáticos/análise , Água do Mar/química , Distribuição Tecidual , Poluentes Químicos da Água/análise
7.
J Assist Reprod Genet ; 36(4): 741-747, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30778819

RESUMO

PURPOSE: Empty follicle syndrome (EFS) is a complex reproductive disorder characterized by the repeated failure to aspirate oocytes from mature ovarian follicles during in vitro fertilization (IVF). In addition to some cases caused by iatrogenic problems and known genetic factors, there are still many unexplained aspects of EFS. Here, we aimed to assess the clinical and genetic characteristics of two EFS patients. METHODS: We have characterized two primary infertility patients with EFS in a nonconsanguineous family from China. Both the patients presented similar clinical phenotypes, that is a few granulosa cells but no oocytes could be retrieved during repeated cycles with normal follicular development, E2 levels, and bioavailable hCG plasma levels. Abnormal oocytes were obtained once or twice between multiple IVF cycles. We performed Sanger sequencing of the LHCGR and ZP1~ZP4 genes in the patients, and further bioinformatics analysis was performed to identify pathogenic elements in the genes. RESULTS: A novel mutation, c.181C>T (p.Arg61Cys), and a known mutation, c.1169_1176delTTTTCCCA (p.Ile390Thrfs*16), in the ZP1 gene were both identified in patient 2, but no mutations were identified in patient 1. The novel mutation inherited from her mother was absent in the control cohort and the ExAc database. The arginine residue is conserved at this position, and its replacement by cysteine was predicted to be deleterious. In another allele, a paternal frameshift mutation was predicted to introduce premature stop codons, resulting in the deletion of 234 amino acids from the C-terminus of the ZP1 protein. CONCLUSIONS: Our findings presented compound heterozygous mutations in ZP1 associated with EFS and abnormal oocytes and provided further new evidence for the genetic basis of EFS and support for the genetic diagnosis of infertile individuals.


Assuntos
Predisposição Genética para Doença , Infertilidade Feminina/genética , Doenças Ovarianas/genética , Glicoproteínas da Zona Pelúcida/genética , Adulto , China/epidemiologia , Feminino , Humanos , Infertilidade Feminina/patologia , Mutação , Oócitos/crescimento & desenvolvimento , Oócitos/patologia , Doenças Ovarianas/patologia , Folículo Ovariano/crescimento & desenvolvimento , Folículo Ovariano/patologia , Ovulação/genética , Indução da Ovulação/métodos , Fenótipo , Zona Pelúcida/patologia
8.
Environ Pollut ; 246: 597-607, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30605815

RESUMO

Di(n-butyl) phthalate (DBP) is extensively used in industrial applications as plasticizer and stabilizer and its presence in the environment may present health risks for human. Previous studies have demonstrated its mutagenic, teratogenic, and carcinogenic ability. However, its effect on mammalian oocyte maturation remains unknown. In this study, we examined the effect of DBP on oocyte maturation both in vitro and in vivo. Our results showed that DBP could significantly reduce mice oocyte germinal vesicle breakdown (GVBD) and polar body extrusion (PBE) rates. In addition, oocyte cytoskeleton was damaged and cortical granule-free domains (CGFDs) were also disrupted. Finally, DBP induced early apoptosis of oocyte and granulosa cells (GCs). Collectively, these data demonstrate that DBP could reduce meiosis competence and mouse oocyte development.


Assuntos
Apoptose/efeitos dos fármacos , Dibutilftalato/toxicidade , Poluentes Ambientais/toxicidade , Meiose/efeitos dos fármacos , Oócitos/efeitos dos fármacos , Animais , Feminino , Células da Granulosa/efeitos dos fármacos , Células da Granulosa/patologia , Humanos , Camundongos , Camundongos Endogâmicos ICR , Oócitos/crescimento & desenvolvimento , Oócitos/patologia
9.
Fertil Steril ; 111(2): 195-196, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30691622

RESUMO

This introduction reviews the background of mitochondrial transplantation. Its goals are to provide both basic and clinical insights into mitochondria and reproduction. The articles will review the biology of mitochondria, mitochondrial DNA, mitochondrial dynamics, and mitochondrial stress (unfolded protein response), discuss novel imaging technology, fluorescent lifetime imaging microscopy to visualize mitochondria and their function in oocytes and early embryos, provide a critical and objective review of the data suggesting and refuting the possible use of mitochondrial DNA as well as other emerging technologies as a biomarker for embryo viability and finally to review the use of mitochondria as a rejuvenation tool in human in vitro fertilization.


Assuntos
Blastocisto/metabolismo , Infertilidade/terapia , Mitocôndrias/metabolismo , Mitocôndrias/transplante , Oócitos/metabolismo , Técnicas de Reprodução Assistida , Animais , Blastocisto/patologia , DNA Mitocondrial/genética , DNA Mitocondrial/metabolismo , Implantação do Embrião , Fertilidade , Humanos , Infertilidade/genética , Infertilidade/metabolismo , Infertilidade/patologia , Mitocôndrias/genética , Mitocôndrias/patologia , Dinâmica Mitocondrial , Oócitos/patologia , Resposta a Proteínas não Dobradas
10.
Fertil Steril ; 111(2): 197-204, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30691623

RESUMO

Mitochondria play a central role in the regulation of energy metabolism in oocytes and preimplantation embryos, where the number and morphology of mitochondria and mitochondrial DNA (mtDNA) content are tightly regulated. A number of mouse models with mitochondrial dysfunction result in infertility, further confirming the key role of mitochondria in female reproductive function. When cells and organisms detect mitochondrial dysfunction they use response mechanisms directed at recovering salvageable mitochondria and eliminating mitochondria that can no longer be rescued. Among these mechanisms, mitochondrial unfolded protein response (UPRmt) has recently been linked with prevention of aging, as compromised mitochondrial stress response contributes to age-related accumulation of damaged proteins, reduced oxidative phosphorylation, and increased reactive oxygen species (ROS) production. These mechanisms seem to be especially relevant for reproduction, as targeted deletion of the UPRmt-regulatory gene Clpp results in female infertility, with impaired oocyte maturation and two-cell embryo development, and failure to form blastocysts. In addition, absence of CLPP results in accelerated depletion of follicles, and a phenotype similar to premature reproductive aging. Further studies will provide novel mechanistic insights for physiologic and pathologic control of oocyte and early embryonic mitochondrial function, which can be exploited for the development of novel therapeutic approaches for the promotion of fertility during the aging process.


Assuntos
Envelhecimento/metabolismo , Blastocisto/metabolismo , Fertilidade , Mitocôndrias/metabolismo , Oócitos/metabolismo , Saúde Reprodutiva , Resposta a Proteínas não Dobradas , Fatores Etários , Envelhecimento/genética , Envelhecimento/patologia , Animais , Blastocisto/patologia , Senescência Celular , Endopeptidase Clp/genética , Endopeptidase Clp/metabolismo , Feminino , Humanos , Infertilidade/genética , Infertilidade/metabolismo , Infertilidade/patologia , Infertilidade/terapia , Masculino , Mitocôndrias/genética , Mitocôndrias/patologia , Dinâmica Mitocondrial , Oócitos/patologia , Técnicas de Reprodução Assistida
11.
Fertil Steril ; 111(2): 212-218, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30691624

RESUMO

Current strategies for embryo assessment in the assisted reproductive technology laboratories rely primarily on morphologic parameters that have limited accuracy for determining embryo viability. Even with the addition of invasive diagnostic interventions such as preimplantation genetic testing for aneuploidy alone or in combination with mitochondrial DNA copy number assessment, at least one third of embryos fail to implant. Therefore, at a time when the clinical benefits of single ET are widely accepted, improving viability assessment of embryos is ever more important. Building on the previous work demonstrating the importance of metabolic state in oocytes and embryos, metabolic imaging via fluorescence lifetime imaging microscopy offers new and potentially useful diagnostic method by detecting natural fluorescence of FAD and NADH, the two electron transporters that play a central role in oxidative phosphorylation. Recent studies demonstrate that fluorescence lifetime imaging microscopy can detect oocyte and embryo metabolic function and dysfunction in a multitude of experimental models and provide encouraging evidence for use in scientific investigation and possibly for clinical application.


Assuntos
Blastocisto/metabolismo , Metabolismo Energético , Fertilização In Vitro , Infertilidade/terapia , Microscopia de Fluorescência , Mitocôndrias/metabolismo , Oócitos/metabolismo , Biomarcadores/metabolismo , Blastocisto/patologia , Sobrevivência Celular , Feminino , Fertilidade , Flavina-Adenina Dinucleotídeo/metabolismo , Humanos , Infertilidade/metabolismo , Infertilidade/patologia , Infertilidade/fisiopatologia , Masculino , Mitocôndrias/patologia , NAD/metabolismo , Oócitos/patologia , Valor Preditivo dos Testes , Gravidez , Resultado do Tratamento
12.
Fertil Steril ; 111(2): 219-226, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30611551

RESUMO

Ovarian aging leads to a decrease in the quantity and quality of oocytes. Aged oocytes have significantly reduced amounts of mitochondria, the energy factories of cells, leading to lower fertilization rates and poor embryonic development. Various techniques have tried to use heterologous or autologous sources of mitochondria to reestablish oocyte health by providing more energy. However, heterologous sources are no longer used owing to the known risk of heteroplasmy. Although autologous methods have recently been tested in humans, they have not shown a clear improvement in embryo quality. In this review, we describe the techniques that have been tested in recent years to provide a state of the art on oocyte rejuvenation through extra injection of mitochondria.


Assuntos
Fertilidade , Infertilidade Feminina/terapia , Mitocôndrias/metabolismo , Mitocôndrias/transplante , Oócitos/metabolismo , Ovário/metabolismo , Rejuvenescimento , Técnicas de Reprodução Assistida , Animais , Fracionamento Celular , Senescência Celular , Feminino , Humanos , Infertilidade Feminina/metabolismo , Infertilidade Feminina/patologia , Infertilidade Feminina/fisiopatologia , Masculino , Idade Materna , Mitocôndrias/patologia , Oócitos/patologia , Ovário/patologia , Ovário/fisiopatologia , Gravidez , Resultado do Tratamento
13.
J Assist Reprod Genet ; 36(3): 425-432, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30610662

RESUMO

Infertility due to Gonadotropin-Resistant Ovary Syndrome (GROS) is a rare type of hypergonadotropic hypogonadism. Here, we report an original case of GROS, associated with compound heterozygous follicle-stimulating hormone receptor (FSHR) variants, in a woman who achieved a live birth by in vitro maturation (IVM) of her oocytes. This 31-year-old woman consulted our assisted reproduction center for a second opinion after having been advised, because of pervasive high serum follicle-stimulating hormone (FSH) levels, to pursue in vitro fertilization (IVF) with donor oocytes. She presented with primary infertility and progressively prolonged menstrual cycles. Her serum FSH levels were indeed found to be high, but in discordance with a normal anti-Müllerian hormone (AMH) level and antral follicle count. Genetic investigation found the patient to be compound heterozygous for two FSHR variants: I160T, a known pathologic variant, and N558H, which has never been previously reported. As there was no ovarian response to high daily doses of exogenous gonadotropins, IVM was proposed to the patient with success and she finally delivered at term a healthy boy. Effects of the receptor variants were analyzed in heterologous cells. Whereas the I160T mutation blocked FSHR membrane trafficking and FSH-stimulated cAMP-dependent signaling in transfected CHO cells, the novel variant, N558H, functioned equivalently to wild-type FSHR in the assays employed. In conclusion, IVM should always be offered as a first-line therapy to infertile women presenting with GROS. The N558H variant discovered in FSHR is novel, but its functional significance, if any, is unresolved and merits further investigation as it may be associated with a recessive FSHR-related disorder.


Assuntos
Hormônio Foliculoestimulante/genética , Técnicas de Maturação in Vitro de Oócitos , Insuficiência Ovariana Primária/genética , Receptores do FSH/genética , Adulto , Animais , Hormônio Antimülleriano/sangue , Células CHO , Cricetulus , Feminino , Hormônio Foliculoestimulante/sangue , Humanos , Nascimento Vivo , Oócitos/crescimento & desenvolvimento , Oócitos/patologia , Insuficiência Ovariana Primária/sangue , Insuficiência Ovariana Primária/patologia , Transdução de Sinais
14.
J Assist Reprod Genet ; 36(3): 433-443, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30627993

RESUMO

PURPOSE: Clinical pregnancy rate after IVF with eSET stagnates between 30 and 40%. In order to increase pregnancy and live birth rates, multiple embryo transfer is still common practice. Providing additional non-invasive tools to choose the competent embryo for transfer could avoid multiple pregnancy and improve time to pregnancy. Cumulus mRNA analysis with quantitative PCR (QPCR) is a non-invasive approach. However, so far, no gene sets have been validated in prospective interventional studies. METHODS: A prospective interventional single-center pilot study with two matched controls (day-3 and day-5 eSET) was performed in 96 patients consenting to the analysis of the cumulus-corona of their oocytes. All patients were super-ovulated for ICSI and eSET at day 3. All oocytes were denuded individually and cumulus was analyzed by quantitative PCR using three predictive genes (EFNB2, SASH1, CAMK1D) and two housekeeping genes (UBC and ß2M). Patients (n = 62) with 2 or more day-3 embryos (good or excellent morphology) had their embryo chosen following the normalized expression of the genes. RESULTS: Corona testing significantly increased the clinical pregnancy and live births rates (63% and 55%) compared to single embryo transfer (eSET) on day 3 (27% and 23%: p < 0.001) and day 5 (43% and 39%: p = 0.022 and p = 0.050) fresh transfer cycle controls with morphology-only selection. Time-to-pregnancy was significantly reduced, regardless of the number of good-quality embryos available on day 3. CONCLUSION: Combining standard morphology scoring and cumulus/corona gene expression analysis increases day-3 eSET results and significantly reduces the time to pregnancy. TRIAL REGISTRATION NUMBER: This is not an RCT study and was only registered by the ethical committee of the University Hospital UZBRUSSEL of the Vrije Universiteit Brussel VUB (BUN: 143201318000).


Assuntos
Células do Cúmulo/patologia , Fertilização In Vitro/métodos , Oócitos/metabolismo , Injeções de Esperma Intracitoplásmicas/métodos , Adulto , Coeficiente de Natalidade , Células do Cúmulo/metabolismo , Embrião de Mamíferos , Feminino , Regulação da Expressão Gênica no Desenvolvimento/genética , Humanos , Estimativa de Kaplan-Meier , Nascimento Vivo , Oócitos/crescimento & desenvolvimento , Oócitos/patologia , Projetos Piloto , Gravidez , Resultado da Gravidez , Taxa de Gravidez , Transferência de Embrião Único/métodos , Tempo para Engravidar
15.
Cell Physiol Biochem ; 51(5): 2341-2358, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30537739

RESUMO

BACKGROUND/AIMS: This study investigated the effect of consecutive superovulation on the ovaries and established a premature ovarian failure (POF) model in mice. METHODS: The mouse POF model was induced by 5-15 consecutive superovulation treatments with pregnant mare serum gonadotropin (PMSG), human chorionic gonadotropin (HCG) and prostaglandin F2α (PGF2α). Normal adult mice were compared with mice displaying natural ovarian aging. The following serum biochemical parameters were measured: including follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone (P), estradiol (E2), inhibin B (INH B), malondialdehyde (MDA), total superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels. Follicles were counted using H&E staining. Levels of 8-hydroxyguanosine (8-OhdG), 4-hydroxynonenal (4-HNE), nitrotyrosine (NTY), anti-Mullerian hormone (AMH) and CDKN2A/ p16 (p16) were detected using immunohistochemical staining. Reactive oxygen species (ROS) levels were measured using dihydroethidium (DHE) staining. Cell apoptosis was detected using an in situ TUNEL fluorescence staining assay. Levels of proteins involved in ROS-related pathways and the p16 protein were detected using Western blotting. Sod1, Sod2 and Sod3 mRNA levels were detected using quantitative polymerase chain reaction (Q-PCR). Oocyte quality was evaluated using in vitro fertilization (IVF) and zygote culture. RESULTS: Consecutive superovulation groups presented lower P, E2, SOD, GSH-Px and INH B levels, significantly higher FSH, LH, MDA and ROS levels, and significantly fewer primordial follicles compared with the control group. Consecutive superovulation groups presented significantly increased levels of Sod2, 8-OhdG, 4-HNE, NTY, significantly increased levels of the SIRT1 and FOXO1 proteins, significantly increased levels of the senescence-associated protein p16, as well as decreased AMH, Sod1 and Sod3 levels and increased granulosa cell apoptosis compared with the control group. CONCLUSION: Consecutive superovulation significantly decreased ovarian function and oocyte quality and increased oxidative stress and apoptosis in the ovary via a mechanism involving the p16 and SIRT1/FOXO1 signaling pathways. These findings suggest that consecutive superovulation may be used to establish a mouse model of ovarian aging.


Assuntos
Ovário/fisiopatologia , Insuficiência Ovariana Primária/sangue , Insuficiência Ovariana Primária/fisiopatologia , Superovulação , Animais , Apoptose , Modelos Animais de Doenças , Feminino , Hormônio Foliculoestimulante/sangue , Humanos , Inibinas/sangue , Hormônio Luteinizante/sangue , Camundongos , Camundongos Endogâmicos C57BL , Oócitos/metabolismo , Oócitos/patologia , Folículo Ovariano/metabolismo , Folículo Ovariano/patologia , Folículo Ovariano/fisiopatologia , Ovário/metabolismo , Ovário/patologia , Estresse Oxidativo , Insuficiência Ovariana Primária/metabolismo , Insuficiência Ovariana Primária/patologia , Progesterona/sangue
16.
Pesqui. vet. bras ; 38(12): 2207-2212, dez. 2018. tab, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-976420

RESUMO

Bovine herpesvirus 1 (BoHV-1) is an important bovine pathogen that is responsible for causing respiratory diseases and reproductive failures. The presence of BoHV-1 in an in vitro embryo production system affects fertilization, maturation, and embryonic development. The objective of this study was to evaluate the developmental capacity of oocytes from naturally infected cows with no reproductive history. Moreover, this study investigated the presence of viral DNA in cumulus oophorus complexes (COCs). Experimental groups were differentiated by titrating the antibodies detected through seroneutralization assays, establishing three groups: seronegative animals (titer lower than 2), low titer (2 to 8), and animals with a titer above or equal to 16. COCs were obtained from 15 donors during 22 sessions of ultrasound-guided follicular aspiration. DNA was extracted from a pool of COCs obtained from all aspirations from the same donor as well as from whole blood and nested PCR reactions were performed. Only COCs with a compact layer of cumulus cells, an intact zona pellucida, and homogeneous cytoplasm were selected for in vitro culture and evaluation of nuclear maturation rate. After culturing for 24 hours, the oocytes were fixed and stained to evaluate the meiotic cell cycle stage. Oocytes that showed a chromosomal configuration in metaphase II were considered to have reached nuclear maturation. Compared with the other groups, the oocyte nuclear maturation rate in animals with a titer greater than or equal to 16 (50%) was compromised (P<0.05). However, the viral titer did not influence the maturation rate of bovine oocytes in animals exhibiting low titration (62.2%) when compared with the control group (76.7%). Viral DNA was not observed in the blood samples but was detected in the COC pool from three seropositive donors. In view of the results obtained, we conclude that natural infections by the BoHV-1 virus can compromise the nuclear maturation rate in cows, depending on the titration levels of antibodies against the virus. Moreover, viral DNA could be present in COCs, contradicting the hypothesis that seropositive animals with no history of clinical symptomatology pose a negligible risk of transmitting BoHV-1 by COCs.(AU)


Herpesvírus bovino 1 (BoHV-1) é um importante patógeno bovino, responsável por causar doenças respiratórias e falhas reprodutivas. A presença do BoHV-1 em sistema de produção in vitro de embriões afeta a fertilização, a maturação e o desenvolvimento embrionário. O objetivo deste estudo foi avaliar a capacidade de desenvolvimento de ovócitos oriundos de vacas infectadas naturalmente sem histórico reprodutivo. Além disso, este estudo investigou a presença do DNA viral em Complexos Cumulus Ooforus (COCs). Os tratamentos foram definidos a partir do título de anticorpos detectados pelos ensaios de soroneutralização, sendo estabelecidos três grupos: animais soronegativos (título menor do que 2), título baixo (2 a 8) e animais com título maior ou igual a 16. Os COCs foram obtidos de 15 doadoras durante 22 sessões de aspiração folicular guiada por ultrassom. A extração do DNA foi realizada em um pool de COCs de todas as aspirações de uma mesma doadora e no sangue total para a realização das reações de Nested-PCR. Para avaliação da taxa de maturação nuclear, foram selecionados para o cultivo in vitro somente os COCs com camada compacta de células do cumulus, zona pelúcida íntegra e citoplasma homogêneo. Após 24 horas de cultivo, os ovócitos foram fixados e corados em lâmina para a avaliação do estádio do ciclo celular meiótico. Os ovócitos que apresentaram configuração cromossômica em metáfase II foram considerados como tendo alcançado a maturação nuclear. Verificou-se comprometimento na taxa de maturação nuclear ovocitária (P<0.05) nos animais de título maior ou igual a 16 (50%). No entanto, não houve influência do título viral na taxa de maturação de ovócitos bovinos em animais que apresentaram titulação baixa (62,2%) quando comparados com o grupo controle (76,7%). O DNA viral não foi identificado nas amostras de sangue, mas foi detectado no pool de COCs de três doadoras soropositivas. Diante dos resultados encontrados conclui-se que vacas infectadas naturalmente pelo vírus BoHV-1 apresentam comprometimento na taxa de maturação nuclear, dependendo do grau de titulação de anticorpos contra o vírus. Ademais, o DNA viral pode estar presente em COCs contrariando a hipótese de que animais sorologicamente positivos e sem histórico de sintomatologia clínica oferecem risco negligível de transmissão do BoHV-1 por COCs.(AU)


Assuntos
Animais , Feminino , Bovinos , Oócitos/patologia , Oócitos/virologia , Infecções por Herpesviridae/veterinária , Herpesvirus Bovino 1 , Rinotraqueíte Infecciosa Bovina , Técnicas de Maturação in Vitro de Oócitos/veterinária
17.
Int J Mol Sci ; 20(1)2018 Dec 26.
Artigo em Inglês | MEDLINE | ID: mdl-30587792

RESUMO

The growth and development of oocyte affect the functional activities of the surrounding somatic cells. These cells are regulated by various types of hormones, proteins, metabolites, and regulatory molecules through gap communication, ultimately leading to the development and maturation of oocytes. The close association between somatic cells and oocytes, which together form the cumulus-oocyte complexes (COCs), and their bi-directional communication are crucial for the acquisition of developmental competences by the oocyte. In this study, oocytes were extracted from the ovaries obtained from crossbred landrace gilts and subjected to in vitro maturation. RNA isolated from those oocytes was used for the subsequent microarray analysis. The data obtained shows, for the first time, variable levels of gene expression (fold changes higher than |2| and adjusted p-value < 0.05) belonging to four ontological groups: regulation of cell proliferation (GO:0042127), regulation of cell migration (GO:0030334), and regulation of programmed cell death (GO:0043067) that can be used together as proliferation, migration or apoptosis markers. We have identified several genes of porcine oocytes (ID2, VEGFA, BTG2, ESR1, CCND2, EDNRA, ANGPTL4, TGFBR3, GJA1, LAMA2, KIT, TPM1, VCP, GRID2, MEF2C, RPS3A, PLD1, BTG3, CD47, MITF), whose expression after in vitro maturation (IVM) is downregulated with different degrees. Our results may be helpful in further elucidating the molecular basis and functional significance of a number of gene markers associated with the processes of migration, proliferation and angiogenesis occurring in COCs.


Assuntos
Apoptose/genética , Proliferação de Células/genética , Oócitos/metabolismo , Transcriptoma , Animais , Movimento Celular/genética , Células do Cúmulo/metabolismo , Células do Cúmulo/patologia , Regulação para Baixo , Feminino , Perfilação da Expressão Gênica , Redes Reguladoras de Genes , Técnicas de Maturação in Vitro de Oócitos , Análise de Sequência com Séries de Oligonucleotídeos , Oócitos/crescimento & desenvolvimento , Oócitos/patologia , RNA/genética , RNA/metabolismo , Suínos , Regulação para Cima
18.
Turk J Med Sci ; 48(4): 750-758, 2018 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-30119150

RESUMO

Background/aim: We aimed to investigate whether oocyte morphologic abnormalities affected cycle outcome in poor responder infertile women who fulfilled the Bologna criteria. Materials and methods: Data were obtained from infertile couples who underwent ICSI and embryo transfer at the Zeynep Kamil Maternity and Children's Health Training and Research Hospital Assisted Reproduction Unit in Istanbul, Turkey. They were allocated to two groups: Group A, poor responders, and Group B, normal responders. All morphological abnormalities of oocytes retrieved were reviewed and grouped as cytoplasmic abnormalities or extracytoplasmic abnormalities. All morphological features were compared between the poor and normal responder groups. ICSI cycle outcomes were described as biochemical pregnancy, fertilization rate, number of top-quality zygotes, top-quality zygote rate, number of embryos transferred, and number of top-quality embryos transferred. The relation between each morphological feature and ICSI outcomes was investigated in each group. Results: The results showed no difference between the groups in terms of morphological features of oocytes. The relation between ICSI cycle outcomes and each oocyte morphological feature was compared in the poor and normal responder groups. Presence of cytoplasmic morphological abnormality was found to significantly correlate with the fertilization rate (P = 0.019) in poor responders. Conclusion: Our data show that oocyte cytoplasmic abnormalities significantly reduce fertilization achievement in poor responders.


Assuntos
Transferência Embrionária , Fertilização , Infertilidade Feminina/patologia , Oócitos/patologia , Injeções de Esperma Intracitoplásmicas , Adulto , Citoplasma/patologia , Feminino , Hospitais , Humanos , Ovulação , Gravidez , Turquia , Zigoto
19.
Syst Biol Reprod Med ; 64(4): 283-290, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29718716

RESUMO

The presence of smooth endoplasmic reticulum aggregates (SERa) in the ooplasm is considered as the most severe oocyte dysmorphism due to its serious and potentially lethal outcomes in offspring. In the present case report, a couple underwent their first intracytoplasmic sperm injection (ICSI) cycle using a gonadotrophin releasing hormone (GnRH) antagonist protocol, followed by fetal ultrasound scanning and amniocentesis. SERa were observed in all oocytes retrieved. A singleton pregnancy was established. The second trimester fetal ultrasound scan revealed a female fetus with overlapping fingers in both hands, and amniocentesis was performed for the detection of chromosomal abnormalities. Comprehensive genetic analysis with the combined use of array-comparative genomic hybridization (CGH), fluoresence in situ hybridization (FISH) and conventional cytogenetics revealed a complex chromosome rearrangement (CCR) involving three break points on two chromosomes, resulting in a reciprocal translocation with a cryptic 2q31 deletion. A week following amniocentesis, there was rupture of amniotic membranes and a stillborn was delivered. This is the first case in the literature to report a CCR with concomitant 2q31 deletion resulting in a well-defined and clinically recognizable contiguous gene syndrome with an abnormal phenotype in a fetus arising from a cohort of oocytes affected by SERa. It is suggested that fertilization and transfer of oocytes with SERa should be avoided, until further research establishes whether there is a causal relationship between the presence of SERa and chromosomal abnormalities in the resulting fetus. ABBREVIATIONS: SER: smooth endoplasmic reticulum; ICSI: intracytoplasmic sperm injection; GnRH: gonadotrophin releasing hormone; CGH: comparative genomic hybridization; FISH: fluoresence in situ hybridization; FSH: follicle stimulating hormone; hCG: human chorionic gonadotrophin; OHSS: ovarian hyperstimulation syndrome; IVF: in vitro fertilization; MII: metaphase II; GV: germinal vesicle; CCR: complex chromosome rearrangement.


Assuntos
Aberrações Cromossômicas , Retículo Endoplasmático Liso/patologia , Oócitos/patologia , Adulto , Feminino , Humanos , Masculino , Injeções de Esperma Intracitoplásmicas
20.
Reproduction ; 155(6): 529-541, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29626105

RESUMO

Male chronic alcohol abuse causes testicular failure and infertility. We analyzed the effects of moderate sub-chronic alcohol intake on sperm morphology, capacitation, fertilization and sperm head decondensation. CF-1 male mice were administered 15% ethanol in drinking water for 15 days; control mice received ethanol-free water. Similar patterns of tyrosine phosphorylation were observed in capacitated spermatozoa of control and treated males. Percentage of hyperactivation (H) and spontaneous (SAR) and progesterone-induced (IAR) acrosome reaction significantly decreased at 120 and 150 min of capacitation in treated males compared to controls (H: 14.1 ± 2.5 vs 23.7 ± 2.6, P < 0.05; SAR-T120 min: 17.9 ± 2.5 vs 32.9 ± 4.1, P < 0.01; IAR-150 min: 43.3 ± 3.5 vs 73.1 ± 1.1, P < 0.001, n = 6). During in vitro fertilization (2.5, 3.5 and 4.5 h post-insemination), there was an increased percentage of fertilized oocytes (with a decondensed sperm head and one or two pronuclei) in treated males (P < 0.001, n = 7). After 60 min of in vitro decondensation with glutathione plus heparin, the percentage of decondensed sperm heads was significantly higher in treated males than in controls (mean ± s.d.: 57.1 ± 5.6 vs 48.3 ± 4.5, P < 0.05, n = 5). The percentage of morphologically normal sperm heads was significantly decreased in treated males with respect to controls (P < 0.001, n = 9). These results show that short-term moderate alcohol consumption in outbred mice affect sperm morphology, hyperactivation, acrosomal exocytosis, and the dynamics of in vitro fertilization and in vitro sperm nuclear decondensation.


Assuntos
Reação Acrossômica/efeitos dos fármacos , Etanol/administração & dosagem , Oócitos/efeitos dos fármacos , Capacitação Espermática/efeitos dos fármacos , Interações Espermatozoide-Óvulo/efeitos dos fármacos , Animais , Anti-Infecciosos Locais/administração & dosagem , Fertilização In Vitro , Masculino , Camundongos , Oócitos/patologia
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