Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 779
Filtrar
1.
Int J Mol Sci ; 22(2)2021 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-33477832

RESUMO

Mammalian reproductive health affects the entire reproductive cycle starting with the ovarian function through implantation and fetal growth. Various environmental and physiological factors contribute to disturbed reproductive health status leading to infertility problems in mammalian species. In the last couple of decades a significant number of studies have been conducted to investigate the transcriptome of reproductive tissues and organs in relation to the various reproductive health issues including endometritis, polycystic ovarian syndrome (PCOS), intrauterine growth restriction (IUGR), preeclampsia, and various age-associated reproductive disorders. Among others, the post-transcriptional regulation of genes by small noncoding miRNAs contributes to the observed transcriptome dysregulation associated with reproductive pathophysiological conditions. MicroRNAs as a class of non-coding RNAs are also known to be involved in various pathophysiological conditions either in cellular cytoplasm or they can be released to the extracellular fluid via membrane-bounded extracellular vesicles and proteins. The present review summarizes the cellular and extracellular miRNAs and their association with the etiology of major reproductive pathologies including PCOS, endometritis, IUGR and age-associated disorders in various mammalian species.


Assuntos
Genitália/metabolismo , MicroRNAs/genética , Reprodução/genética , Saúde Reprodutiva , Animais , Implantação do Embrião/genética , Feminino , Regulação da Expressão Gênica/genética , Redes Reguladoras de Genes/genética , Humanos , Síndrome do Ovário Policístico/genética , Síndrome do Ovário Policístico/patologia , Gravidez
2.
Gene ; 766: 145117, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-32920039

RESUMO

The extracellular vesicles (EVs) of uterine flushing fluids (UFs) mediate intrauterine communication between conceptus and uterus in pigs. The small RNAs of UFs-EVs are widely recognized as important factors that influence embryonic implantation. However, small RNAs expression profiles of porcine UFs-EVs during peri-implantation are still unknown. In this study, cup-shaped EVs of porcine UFs on days 10 (D10), 13 (D13) and 18 (D18) of pregnancy were isolated and characterized. The expression of small RNAs in these EVs was comprehensively profiled through sequencing. A total of 152 known microRNAs (miRNAs), 43 novel miRNAs, 6248 known Piwi-interacting RNAs (piRNAs) and 110 novel piRNAs were identified. Among these small RNAs, RT-qRCR results indicated that ssc-let-7f-5p, ssc-let-7i-5p and ssc-let-7g were differentially expressed during the three stages. Bioinformatics analysis showed that the miRNAs differentially expressed in the three comparisons (D10 vs D13, D13 vs D18 and D10 vs D18) were involved in important processes and pathways related to immunization, endometrial receptivity and embryo development, which play important roles in embryonic implantation. Our results reveal that EVs from porcine UFs contain various small RNAs with potentially vital effects on implantation. This research also provides resources for studies of miRNAs and piRNAs in the cross-talk between embryo and endometrium.


Assuntos
Implantação do Embrião/genética , Vesículas Extracelulares/genética , MicroRNAs/genética , Útero/fisiologia , Animais , Desenvolvimento Embrionário/genética , Endométrio/fisiologia , Feminino , Gravidez , RNA Interferente Pequeno/genética , RNA-Seq/métodos , Suínos
3.
Nat Commun ; 11(1): 3987, 2020 08 10.
Artigo em Inglês | MEDLINE | ID: mdl-32778678

RESUMO

Aneuploidy, the presence of an abnormal number of chromosomes, is a major cause of early pregnancy loss in humans. Yet, the developmental consequences of specific aneuploidies remain unexplored. Here, we determine the extent of post-implantation development of human embryos bearing common aneuploidies using a recently established culture platform. We show that while trisomy 15 and trisomy 21 embryos develop similarly to euploid embryos, monosomy 21 embryos exhibit high rates of developmental arrest, and trisomy 16 embryos display a hypo-proliferation of the trophoblast, the tissue that forms the placenta. Using human trophoblast stem cells, we show that this phenotype can be mechanistically ascribed to increased levels of the cell adhesion protein E-CADHERIN, which lead to premature differentiation and cell cycle arrest. We identify three cases of mosaicism in embryos diagnosed as full aneuploid by pre-implantation genetic testing. Our results present the first detailed analysis of post-implantation development of aneuploid human embryos.


Assuntos
Aneuploidia , Implantação do Embrião/genética , Embrião de Mamíferos , Desenvolvimento Embrionário , Antígenos CD/genética , Caderinas/genética , Caderinas/metabolismo , Adesão Celular , Pontos de Checagem do Ciclo Celular , Linhagem da Célula , Segregação de Cromossomos , Cromossomos Humanos Par 16 , Cromossomos Humanos Par 21 , Feminino , Genes erbB-1/genética , Testes Genéticos , Humanos , Monossomia , Mosaicismo , Gravidez , Células-Tronco , Trissomia
4.
Nat Commun ; 11(1): 2782, 2020 06 03.
Artigo em Inglês | MEDLINE | ID: mdl-32493987

RESUMO

The transcriptional repressor Blimp1 controls cell fate decisions in the developing embryo and adult tissues. Here we describe Blimp1 expression and functional requirements within maternal uterine tissues during pregnancy. Expression is robustly up-regulated at early post-implantation stages in the primary decidual zone (PDZ) surrounding the embryo. Conditional inactivation results in defective formation of the PDZ barrier and abnormal trophectoderm invasion. RNA-Seq analysis demonstrates down-regulated expression of genes involved in cell adhesion and markers of decidualisation. In contrast, genes controlling immune responses including IFNγ are up-regulated. ChIP-Seq experiments identify candidate targets unique to the decidua as well as those shared across diverse cell types including a highly conserved peak at the Csf-1 gene promoter. Interestingly Blimp1 inactivation results in up-regulated Csf1 expression and macrophage recruitment into maternal decidual tissues. These results identify Blimp1 as a critical regulator of tissue remodelling and maternal tolerance during early stages of pregnancy.


Assuntos
Decídua/metabolismo , Fator 1 de Ligação ao Domínio I Regulador Positivo/metabolismo , Transcrição Genética , Animais , Decídua/ultraestrutura , Ectoderma/metabolismo , Ectoderma/ultraestrutura , Implantação do Embrião/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Fator Estimulador de Colônias de Macrófagos/genética , Fator Estimulador de Colônias de Macrófagos/metabolismo , Macrófagos/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Mutação/genética , Gravidez , Regiões Promotoras Genéticas , Trofoblastos/metabolismo , Trofoblastos/ultraestrutura , Regulação para Cima/genética
5.
Development ; 147(6)2020 03 17.
Artigo em Inglês | MEDLINE | ID: mdl-32184271

RESUMO

Reproductive decline in older female mice can be attributed to a failure of the uterus to decidualise in response to steroid hormones. Here, we show that normal decidualisation is associated with significant epigenetic changes. Notably, we identify a cohort of differentially methylated regions (DMRs), most of which gain DNA methylation between the early and late stages of decidualisation. These DMRs are enriched at progesterone-responsive gene loci that are essential for reproductive function. In female mice nearing the end of their reproductive lifespan, DNA methylation fidelity is lost at a number of CpG islands (CGIs) resulting in CGI hypermethylation at key decidualisation genes. Importantly, this hypermethylated state correlates with the failure of the corresponding genes to become transcriptionally upregulated during the implantation window. Thus, age-associated DNA methylation changes may underlie the decidualisation defects that are a common occurrence in older females. Alterations to the epigenome of uterine cells may therefore contribute significantly to the reproductive decline associated with advanced maternal age.


Assuntos
Envelhecimento/genética , Implantação do Embrião/genética , Epigênese Genética/fisiologia , Reprodução/fisiologia , Animais , Células Cultivadas , Ilhas de CpG/genética , Metilação de DNA/fisiologia , Decídua/fisiologia , Embrião de Mamíferos , Feminino , Masculino , Idade Materna , Camundongos , Camundongos Endogâmicos C57BL , Gravidez , Reprodução/genética
6.
Reprod Sci ; 27(1): 29-38, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-32046408

RESUMO

Recurrent implantation failure (RIF) is defined when pregnancy failure occurs after two consecutive in vitro fertilization-embryo transfers to the endometrium using at least four high-quality embryos in women. MicroRNAs are well-known function modulators and are involved in many diseases. Recently, studies on microRNA and recurrent pregnancy loss (RPL) have been actively carried out; however, single nucleotide polymorphisms of miRNA in RPL are not well known. Therefore, we set the aim of this study to identify whether polymorphisms in miRNAs that miR-27aA>G, miR-423C>A, miR-449bA>G, and miR-604A>G are risk factors for idiopathic recurrent implantation failure (RIF) in Korean women. Genotyping was assessed with a polymerase chain reaction-restriction fragment length polymorphism assay. We examined polymorphisms in four miRNA genes: miR-27aA>G, miR-423C>A, miR-449bA>G, and miR-604A>G. We found that the miR-27aA>G, miR-449bA>G, and miR-604A>G polymorphisms were significantly associated with a risk of RIF. In addition, the miR-27aA>G and miR-449bA>G polymorphisms were associated with the frequency of implantation failures. Specifically, the miR-449bAG+GG genotype was associated with RIF prevalence (total RIF: adjusted odd ratio [AOR] = 1.584, 95% CI = 1.008-2.490, P = 0.046; IF ≥ 3 group: AOR = 1.747, 95% CI = 1.088-2.803, P = 0.021; IF ≥ 4: AOR = 1.932, 95% CI = 1.122-3.327, P = 0.018). Based on these results, the miR-449b A>G may be a predisposing factor to RIF susceptibility. However, the mechanism underlying the function of miR-449b A>G in RIF remains to be determined and further studies are needed to improve understanding of the roles of miR-449b A>G, using a larger and more heterogeneous cohort.


Assuntos
Aborto Habitual/genética , Implantação do Embrião/genética , Predisposição Genética para Doença , MicroRNAs/genética , Polimorfismo de Nucleotídeo Único , Feminino , Genótipo , Humanos
7.
Mol Biol Rep ; 47(3): 1751-1758, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32006196

RESUMO

In vitro fertilization failure is not only the cause of despair among couples and individuals undergoing the treatment, it has also been contributing to the impediment of assistive reproductive technologies' development. MicroRNAs (miRNAs) have been linked to significant events in the reproduction course. The identification of miRNA polymorphisms may provide a good lead for the potential of diagnosis and treatment of unidentified in vitro fertilization (IVF) failure causes. The aim of our study is to explore the association between miRNA polymorphisms (mir-320b T>C and mir-27a G >A) and IVF failure. Our case-control study consisted of 200 Kurdish women in total, 100 with IVF failure and the other 100 control who have had at least two successful pregnancies and no history of pregnancy loss, we used tetra amplification refractory mutation system PCR to identify the polymorphisms within the groups. The TT genotype of mir-320b was found more frequently in IVF failure patients when compared to the healthy women (OR 8.07, CI 2.18-29.78, P = 0.001) and T allele was more present in the case group (OR 1.83, CI 91.04-2.12, P = 0.034), however mir-27a seemed to show no association with IVF failure in regards to genotype and allele frequencies. The difference in genotype and allele frequencies of mir-320b of the two groups may indicate that it has an effect on the target mRNAs and alter the implantation of embryo during IVF cycles.


Assuntos
Fertilização In Vitro/métodos , MicroRNAs/genética , Polimorfismo de Nucleotídeo Único , Adulto , Alelos , Sequência de Bases , Estudos de Casos e Controles , Implantação do Embrião/genética , Feminino , Frequência do Gene , Estudos de Associação Genética/métodos , Genótipo , Humanos , Fatores de Risco , Adulto Jovem
8.
Biomed Res Int ; 2020: 6279795, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32104701

RESUMO

This review analyses the genetic mechanisms of acephalic spermatozoa (AS) defects, which are associated with primary infertility in men. Several target genes of headless sperms have been identified but intracytoplasmic sperm injection (ICSI) outcomes are complex. Based on electron microscopic observations, broken points of the sperm neck are AS defects that are based on various genes that can be classified into three subtypes: HOOK1, SUN5, and PMFBP1 genes of subtype II; TSGA10 and BRDT genes of subgroup III, while the genetic mechanism(s) and aetiology of AS defects of subtype I have not been described and remain to be explored. Interestingly, all AS sperm of subtype II achieved better ICSI outcomes than other subtypes, resulting in clinical pregnancies and live births. For subtype III, the failure of clinical pregnancy can be explained by the defects of paternal centrioles that arrest embryonic development; for subtype I, this was due to a lack of a distal centriole. Consequently, the embryo quality and potential ICSI results of AS defects can be predicted by the subtypes of AS defects. However, this conclusion with regard to ICSI outcomes based on subtypes still needs further research, while the existence of quality of oocyte and implantation failure in women cannot be ignored.


Assuntos
Infertilidade Masculina/genética , Oócitos/metabolismo , Espermatozoides/patologia , Adulto , Implantação do Embrião/genética , Implantação do Embrião/fisiologia , Feminino , Humanos , Infertilidade Masculina/patologia , Masculino , Proteínas de Membrana/genética , Proteínas Associadas aos Microtúbulos/genética , Oócitos/crescimento & desenvolvimento , Oócitos/patologia , Gravidez , Taxa de Gravidez , Espermatozoides/crescimento & desenvolvimento
9.
Reprod Domest Anim ; 55(4): 523-529, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31986225

RESUMO

Pregnancy is a complex process in which significant changes occur continually in both the corpora lutea and in the endometrium of the females and varies depending on the embryonic, pre-implantation or foetal stages. In the embryonic stages, the majority of genes expressed in the pig embryo correspond to the loss of cellular pluripotency. In contrast, the implantation consists of three phases: elongation of the conceptus, adhesion and union of the embryo to the endometrial epithelium. During these phases, many factors are expressed, including growth factors, molecules that facilitate adhesion and cytokines. All these changes are ultimately regulated by different lipid and hormonal substances, specifically by progesterone, oestradiol and prostaglandins, which regulate the expression of many proteins necessary for the development of the embryo, endometrial remodelling and embryo-maternal communication. This paper is a review of primary gene regulatory mechanisms in pigs during different stages of implantation.


Assuntos
Embrião de Mamíferos/metabolismo , Desenvolvimento Embrionário/genética , Regulação da Expressão Gênica no Desenvolvimento , Gravidez/fisiologia , Animais , Implantação do Embrião/genética , Endométrio/metabolismo , Feminino , Troca Materno-Fetal , Suínos
10.
Sci Rep ; 10(1): 854, 2020 01 21.
Artigo em Inglês | MEDLINE | ID: mdl-31965014

RESUMO

In the pre-implantation embryo, aneuploidy resulting from chromosome segregation error is considered responsible for pregnancy loss. However, only a few studies have examined the relationship between chromosome segregation errors during early cleavage and development. Here, we evaluated this relationship by live-cell imaging using the histone H2B-mCherry probe and subsequent single blastocyst transfer using mouse embryos obtained by in vitro fertilization. We showed that some embryos exhibiting early chromosomal segregation error and formation of micronuclei retained their developmental potential; however, the error affected the blastocyst/arrest ratio. Further, single-cell sequencing after live-cell imaging revealed that all embryos exhibiting micronuclei formation during 1st mitosis showed aneuploidy at the 2-cell stage. These results suggest that early chromosome segregation error causing micronuclei formation affects ploidy and development to blastocyst but does not necessarily cause developmental failure after the blastocyst stage. Our result suggests the importance of the selection of embryos that have reached blastocysts.


Assuntos
Blastocisto , Segregação de Cromossomos , Implantação do Embrião/genética , Embrião de Mamíferos , Desenvolvimento Embrionário/genética , Aneuploidia , Animais , Transferência Embrionária , Feminino , Fertilização In Vitro , Camundongos , Camundongos Endogâmicos ICR , Gravidez
11.
J Assist Reprod Genet ; 37(1): 33-43, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31758513

RESUMO

PURPOSE: High progesterone is associated with low implantation rate. Our previous study demonstrated that DNA methylation in endometrium was increased in women with high progesterone in IVF cycles. However, the DNA methylation status is still not yet confirmed, and how it affects endometrial receptivity in high progesterone is still unknown. Current study investigated the effects of high progesterone on DNA methylation and gene expression of adhesion molecules on endometrium during implantation window. METHODS: A cohort study included 20 women with high progesterone (HP) and 20 with normal progesterone (NP) on the day of human chorionic gonadotropin (hCG) administration after controlled ovarian hyperstimulation in IVF cycle. Endometrial tissues were collected on the 7th day after hCG administration. Immunohistochemical staining of DNA methyltransferases (DNMT1 and DNMT3B) and adhesion molecules (MUC1, CDH1 and CTNNB1) were performed. Methylation of MUC1, CDH1, and CTNNB1 promoter regions was detected by Sequenom MassARRAY or bisulfite sequencing PCR. RT-qPCR was used to quantify mRNA expression levels, and correlation of methylation and gene expression level of the adhesion molecules were determined. RESULTS: DNMT3B, but not DNMT1, in nucleus of luminal and glandular epithelial cells in HP group was significantly higher than that in NP group. Promoter regions of CDH1 and CTNNB1, but not MUC1, in endometrium of HP group were hypermethylated. Protein and mRNA expression of MUC1, CDH1, and CTNNB1 in endometrium of HP group was significantly lower than that in NP group. Level of DNA methylation was negatively correlated with the gene expression of CDH1 and CTNNB1, but not MUC1. CONCLUSIONS: DNA hypermethylation and low expression of adhesion molecules on endometrium were associated with high progesterone during implantation window, which may contribute to the underlying epigenetic mechanism in the failure of IVF treatment.


Assuntos
Moléculas de Adesão Celular/metabolismo , Metilação de DNA , Implantação do Embrião/genética , Endométrio/metabolismo , Fertilização In Vitro/métodos , Infertilidade Feminina/metabolismo , Progesterona/farmacologia , Adulto , Moléculas de Adesão Celular/genética , Gonadotropina Coriônica/administração & dosagem , Estudos de Coortes , Implantação do Embrião/efeitos dos fármacos , Endométrio/efeitos dos fármacos , Epigênese Genética , Feminino , Perfilação da Expressão Gênica , Humanos , Infertilidade Feminina/tratamento farmacológico , Infertilidade Feminina/genética , Indução da Ovulação/métodos , Progestinas/farmacologia
12.
J Mol Endocrinol ; 64(1): 43-52, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31786540

RESUMO

Receptive endometrium is a prerequisite for successful embryo implantation, and it follows that poor endometrial receptivity is a leading cause of implantation failure. miRNAs play important roles as epigenetic regulators of endometrial receptivity and embryo implantation through post-transcriptional modifications. However, the mechanisms of action of many miRNAs are poorly understood. In this study, we investigated the role of the miR-183 family, comprising three miRNAs (miR-183-5p, miR-182-5p, and miR-96-5p) in endometrial receptivity and embryo implantation. The miR-183 family shows estrogen-dependent upregulation in endometrial Ishikawa (IK) cells. The miR-183 family also has a positive role in migration and proliferation of IK cells. Furthermore, JAr spheroid attachment experiments show that attachment rates were significantly decreased after treatment of IK cells with inhibitors for miR-183-5p and miR-182-5p and increased after treatment with miR-183-5p-mimic and miR-96-5p-mimic, respectively. The downstream analysis shows that catenin alpha 2 (CTNNA2) is a potential target gene for miR-183-5p, and this was confirmed in luciferase reporter assays. An in vivo mouse pregnancy model shows that inhibition of miR-183-5p significantly decreases embryo implantation rates and increases CTNNA2 expression. Downregulation of CTNNA2 in endometrial cells by miR-183-5p may be significant in mediating estrogenic effects on endometrial receptivity. In conclusion, miR-183-5p and the CTNNA2 gene may be potential biomarkers for endometrial receptivity and may be useful diagnostic and therapeutic targets for successful embryo implantation.


Assuntos
Implantação do Embrião/genética , MicroRNAs/genética , Útero/fisiologia , Animais , Biomarcadores/metabolismo , Movimento Celular/genética , Proliferação de Células/genética , Células Cultivadas , Regulação para Baixo/genética , Implantação do Embrião/fisiologia , Endométrio/fisiologia , Feminino , Células HEK293 , Humanos , Masculino , Camundongos , Camundongos Endogâmicos ICR , Gravidez , alfa Catenina/genética
13.
J Anim Breed Genet ; 137(2): 123-138, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31657065

RESUMO

Uterine capacity (UC), defined as the total number of kits from unilaterally ovariectomized does at birth, has a high genetic correlation with litter size. The aim of our research was to identify genomic regions associated with litter size traits through a genomewide association study using rabbits from a divergent selection experiment for UC. A high-density SNP array (200K) was used to genotype 181 does from a control population, high and low UC lines. Traits included total number born (TNB), number born alive (NBA), number born dead, ovulation rate (OR), implanted embryos (IE) and embryo, foetal and prenatal survivals at second parity. We implemented the Bayes B method and the associations were tested by Bayes factors and the percentage of genomic variance (GV) explained by windows. Different genomic regions associated with TNB, NBA, IE and OR were found. These regions explained 7.36%, 1.27%, 15.87% and 3.95% of GV, respectively. Two consecutive windows on chromosome 17 were associated with TNB, NBA and IE. This genomic region accounted for 6.32% of GV of TNB. In this region, we found the BMP4, PTDGR, PTGER2, STYX and CDKN3 candidate genes which presented functional annotations linked to some reproductive processes. Our findings suggest that a genomic region on chromosome 17 has an important effect on litter size traits. However, further analyses are needed to validate this region in other maternal rabbit lines.


Assuntos
Genoma/genética , Tamanho da Ninhada de Vivíparos/genética , Coelhos/genética , Seleção Genética , Animais , Mapeamento Cromossômico/veterinária , Implantação do Embrião/genética , Feminino , Estudo de Associação Genômica Ampla/veterinária , Genótipo , Desequilíbrio de Ligação , Nascimento Vivo/genética , Nascimento Vivo/veterinária , Ovulação/genética , Fenótipo , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Coelhos/fisiologia
14.
Eur J Med Genet ; 63(2): 103741, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31445143

RESUMO

OBJECTIVE: To determine the pregnancy outcome potential of euploid, mosaic and aneuploid embryos. DESIGN: Retrospective study. SETTING: Reference genetics laboratories. PATIENT(S): 2654 PGT-A cycles with euploid characterized embryo transfers, 253 PGT-A cycles with transfer of embryos characterized as mosaic, and 10 PGT-A cycles with fully abnormal embryo transfers. INTERVENTION(S): Blastocysts were assessed by trophectoderm (TE) biopsy followed by PGT-A via array CGH or NGS. MAIN OUTCOME MEASURE(S): Implantation, miscarriage, ongoing implantation rates (OIR), and karyotype if available, were compared between different embryo groups, and between the two PGT-A techniques. RESULTS: The Ongoing Pregnancy Rate (OPR)/transfer was significantly higher for NGS-classified euploid embryos (85%) than for aCGH ones (71%) (p < 0.001), but the OPR/cycle was similar (63% vs 59%). NGS-classified mosaic embryos resulted in 37% OPR/cycle (p < 0.001 compared to euploid). Mosaic aneuploid embryos with <40% abnormal cells in the TE sample had an OIR of 50% compared to 27% for mosaics with 40-80% abnormal cells in the TE, and 9% for complex mosaic embryos. All the karyotyped ongoing pregnancies (n = 29) were euploid. Transfers of embryos classified as aneuploid via aCGH (n = 10) led to one chromosomally abnormal pregnancy. CONCLUSION(S): NGS-classified euploid embryos yielded higher OIRs but similar OPRs/cycle compared to aCGH. NGS-classified mosaic embryos had reduced potential to reach term, compared to euploid embryos. If they did reach term, those with karyotype results available were euploid. Embryos carrying uniform aneuploidies affecting entire chromosomes were mostly unable to implant after transfer, and the one that implanted ended up in a chromosomally abnormal live birth.


Assuntos
Implantação do Embrião/genética , Transferência Embrionária , Testes Genéticos , Mosaicismo/embriologia , Resultado da Gravidez , Diagnóstico Pré-Implantação , Aborto Espontâneo/genética , Adulto , Aneuploidia , Blastocisto/patologia , Hibridização Genômica Comparativa , Ectoderma , Feminino , Feto/anormalidades , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Cariotipagem , Nascimento Vivo , Gravidez , Estudos Retrospectivos
15.
Eur J Med Genet ; 63(2): 103731, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31362121

RESUMO

The clinical application of a PGT-A program implementing single euploid embryo transfer is evaluated over a 6.5 year period, beginning with its early validation phases. Euploidy embryo status is inversely correlated to oocyte source age and positively correlated to blastocyst quality grades. However, once a single euploid embryo is transferred, high levels of implantation and live birth success are attained independent of patient age and embryo quality, with only AA blastocysts exhibiting improved implantation. Factors influencing successful outcomes are discussed, including the management of mosaic NGS profiles. Overall, distinct advantages to a dedicated PGT-A/single euploid embryo transfer program are clearly evident in per cycle start comparisons to control cycles and national average statistics by age groups.


Assuntos
Aneuploidia , Implantação do Embrião/genética , Fertilização In Vitro , Testes Genéticos/métodos , Diagnóstico Pré-Implantação/métodos , Adulto , Blastocisto/citologia , Blastocisto/fisiologia , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Nascimento Vivo , Mosaicismo , Oócitos/citologia , Oócitos/fisiologia , Gravidez
16.
Endocrinology ; 161(1)2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-31875883

RESUMO

Uterine receptivity is critical for establishing and maintaining pregnancy. For the endometrium to become receptive, stromal cells must differentiate into decidual cells capable of secreting factors necessary for embryo survival and placental development. Although there are multiple reports of autophagy induction correlated with endometrial stromal cell (ESC) decidualization, the role of autophagy in decidualization has remained elusive. To determine the role of autophagy in decidualization, we utilized 2 genetic models carrying mutations to the autophagy gene Atg16L1. Although the hypomorphic Atg16L1 mouse was fertile and displayed proper decidualization, conditional knockout in the reproductive tract of female mice reduced fertility by decreasing the implantation rate. In the absence of Atg16L1, ESCs failed to properly decidualize and fewer blastocysts were able to implant. Additionally, small interfering RNA knock down of Atg16L1 was detrimental to the decidualization response of human ESCs. We conclude that Atg16L1 is necessary for decidualization, implantation, and overall fertility in mice. Furthermore, considering its requirement for human endometrial decidualization, these data suggest Atg16L1 may be a potential mediator of implantation success in women.


Assuntos
Proteínas Relacionadas à Autofagia/genética , Autofagia/genética , Decídua/metabolismo , Endométrio/metabolismo , Mutação , Animais , Proteínas Relacionadas à Autofagia/metabolismo , Diferenciação Celular/genética , Células Cultivadas , Decídua/citologia , Implantação do Embrião/genética , Endométrio/citologia , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Interferência de RNA , Células Estromais/citologia , Células Estromais/metabolismo
17.
Exp Cell Res ; 388(1): 111718, 2020 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-31874176

RESUMO

Successful implantation happens only when the development of a competent blastocyst synchronized with the differentiation of a receptive uterus. The exact mechanism affecting embryo implantation competency is still unclear. Previous data from our laboratory showed that several members of the let-7 family were up-regulated in the implanting dormant blastocysts and prohibited embryo activation through down-regulation integrin-ß3. However, how the mir-let-7 family is regulated is still a question. In this study, the in vitro co-culture model was applied to imitate implantation. Human embryo surrogate Jeg-3 spheroids and endometrium epithelial cells Ishikawa were used. The following views were demonstrated. Firstly,Wnt/ß-catenin signaling is essential for Jeg-3 spheroids implantation. Secondly, mir-let-7a is repressed by Wnt signaling, and low let-7a is beneficial for spheroids attachment and outgrowth. Third, in contrast with let-7a, lin28a is up-regulated by Wnt and promotes attachment and outgrowth. Lastly, the function of Wnt in embryo surrogate spheroids in implantation is mediated through lin28a/let-7a axis. In summary, our findings suggest Wnt/ß-catenin signaling strength human embryo surrogate spheroids implanting potential through regulation lin28a/let-7a axis.


Assuntos
Endométrio/citologia , MicroRNAs/metabolismo , Proteínas de Ligação a RNA/genética , Esferoides Celulares/metabolismo , Trofoblastos/citologia , Via de Sinalização Wnt , Linhagem Celular Tumoral , Técnicas de Cocultura , Implantação do Embrião/genética , Endométrio/metabolismo , Células Epiteliais/metabolismo , Feminino , Humanos , MicroRNAs/genética , Proteínas de Ligação a RNA/metabolismo , Esferoides Celulares/citologia , Trofoblastos/metabolismo
18.
Am J Reprod Immunol ; 83(1): e13196, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31595580

RESUMO

PROBLEM: DNA methylation profile in mid-secretory phase of endometrium is reported to be varied from other phases in natural menstrual cycle. Therefore, we intended to study the impairment in endometrial receptivity by performing whole-genome methylation and gene expression profiling in endometrium of recurrent implantation failure patients (RIF) during IVF under controlled ovarian stimulation (COS). METHOD OF STUDY: Endometrial biopsies were collected from IVF-RIF patients (cases, n = 6) and healthy fertile oocyte donors (controls, n = 6) undergoing COS after 6/7th day of human chorionic gonadotropin administration. The whole-genome methylation and gene expression microarray were performed and analysed by GenomeStudio software (P < .05 by Illumina Custom Model), whereas the enrichment analysis was performed using "Database for Annotation, Visualization and Integrated Discovery" (DAVID, V6.8). Significant differentially methylated genes were correlated with dys-regulated genes using Pearson's correlation. RESULTS: Differential methylation in RIF patients revealed 448 CpG sites. The enrichment analysis showed aberrant methylation in genes involved in immunological response and G protein activity. Methylation in NLRP2 gene in inflammatory pathway had significant negative correlation with gene expression (P = .008), whereas SERPINA5 gene that is already known to be involved in endometrial receptivity was observed to be hypomethylated in promoter region with highest delta beta value and up-regulated in gene expression analysis. CONCLUSION: The aberrant methylation of genes involved in immunological functions and G protein activation was found to be prevalent which might suggest a role in endometrial receptivity. However, the findings need to be further validated on a larger cohort of IVF-RIF patients.


Assuntos
Metilação de DNA , Implantação do Embrião/genética , Implantação do Embrião/imunologia , Endométrio/fisiologia , Fertilização In Vitro , Adulto , Gonadotropina Coriônica/uso terapêutico , Feminino , Humanos , Transcriptoma , Falha de Tratamento , Adulto Jovem
19.
Theriogenology ; 142: 196-206, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31606658

RESUMO

The maternal endometrium undergoes transformations during early pregnancy period to regulate the paracellular permeability across the epithelium and to enable adhesion between the trophoblast and endometrial epithelial cells. These transformations, under the influence of ovarian hormones, are associated with a partial loss in polarity of epithelial cell that is regulated by tight junctions (TJ), adherens junctions (AJ) and associated polarity protein complexes. This study examined the change in expression and distribution of proteins associated with TJs, AJs and apical partition defective (PAR) complex in porcine endometrium on Days 10, 13 and 16 of estrous cycle and pregnancy. Moreover, effect of hormones, progesterone (P4) and 17-ß estradiol (E2) on polar phenotype of endometrial epithelial cells was also investigated in vitro. There was pregnancy induced increase in gene and protein expression of TJ associated claudin-1 (CLDN1) on Day 13 of pregnancy as compared to corresponding day of estrous cycle and a decrease in TJ protein, zona occludens-1 (ZO-1) and PAR complex associated PAR6 expression levels on Day 16 of pregnancy (P < 0.05). Immunofluorescence studies revealed that on Days 10 and 13, TJ proteins occludin (OCLN) and ZO-1were primarily present in the apical region of lateral epithelial membrane. On Day 16 of pregnancy, whereas, OCLN redistributed into cytoplasm, ZO-1 decreased apically but was found to localize in the basal epithelium. The AJ proteins cadherin and ß-catenin were located at the apical epithelium on Day 10 of estrous cycle and pregnancy and Day 13 of estrous cycle. On Days 13 and 16 of pregnancy both proteins were expressed in the lateral membrane and co-localization between these proteins was observed on Day 16. On Day 10, PAR complex proteins PAR3, cell division control protein 42 (CDC42) and atypical protein kinase C (aPKC) ζ were observed in apical epithelium and in lateral membrane and CDC42 was also present in the cytoplasm of epithelium. Pregnancy induced redistribution of aPKCζ to cytoplasm and CDC42 to apical surface of luminal epithelium was observed on Days 13 and 16. The in vitro P4 and E2 treatment of epithelial cells mimicked in vivo results. These results indicate that P4 and E2 regulate alterations in epithelium that may facilitate embryo implantation and given the role of cadherin, catenin and CDC42 in embryo invasion, change in distribution of these proteins may limit the invasiveness of porcine conceptuses into the stroma.


Assuntos
Polaridade Celular/genética , Endométrio/metabolismo , Moléculas de Adesão Juncional/genética , Moléculas de Adesão Juncional/metabolismo , Prenhez , Suínos , Junções Aderentes/genética , Junções Aderentes/metabolismo , Animais , Células Cultivadas , Implantação do Embrião/genética , Feminino , Expressão Gênica , Idade Gestacional , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Gravidez , Prenhez/genética , Prenhez/metabolismo , Suínos/embriologia , Suínos/genética , Suínos/metabolismo , Junções Íntimas/genética , Junções Íntimas/metabolismo , Distribuição Tecidual
20.
Hum Genomics ; 13(1): 68, 2019 12 16.
Artigo em Inglês | MEDLINE | ID: mdl-31842980

RESUMO

BACKGROUND: Recurrent implantation failure (RIF) is the failure of embryos to implant more than two times in a given individual. There is debate about a precise definition for RIF, but we consider more than two implantation failures for individuals who undergo in vitro fertilization-embryo transfer (IVF-ET) to constitute RIF. There are many potential reasons for RIF, including embryonic factors, immunological factors, uterine factors, coagulate factors, and genetic factors. Genetic variation has been suggested as one of the contributing factors leading to RIF, and a number of single-nucleotide polymorphisms (SNPs) have been reported to be associated with RIF. The recent elucidation of miRNA functions has provided new insight into the regulation of gene expression. METHODS: We investigated associations between polymorphisms in four miRNAs and RIF in 346 Korean women: 118 patients with RIF and 228 controls. We determined the genotypes of the miRNAs in the study participants by polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) analysis. We analyzed the effects of genotypes, allele combinations, and environmental and clinical factors on the risk of RIF. RESULTS: The miR-25 T/miR-125aT/miR-222G (odds ratio (OR), 0.528; 95% confidence interval (CI), 0.282-0.990; P = 0.044) and miR-25 T/miR-125aT allele combinations were associated with a reduced risk of RIF. The miR-25 T/miR-32C/miR-125aC/miR-222 T allele combination was associated with an increased risk of RIF. The miR-222GT+TT genotypes interacted with high prothrombin time (≥ 12 s) to increase the risk of RIF. CONCLUSIONS: MicroRNA polymorphisms are significantly different between patients that experience RIF and healthy controls. Combinations of microRNA polymorphisms were associated with the risk of RIF. Interactions between environmental factors and genotypes increased the risk of RIF in Korean women.


Assuntos
Implantação do Embrião/genética , Estudos de Associação Genética , Predisposição Genética para Doença , MicroRNAs/genética , Polimorfismo de Nucleotídeo Único/genética , Adulto , Alelos , Fatores de Coagulação Sanguínea/metabolismo , Estudos de Casos e Controles , Feminino , Humanos , MicroRNAs/metabolismo , Gravidez
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...