RESUMO
BACKGROUND: Epigenetic abnormalities caused by superovulation have recently attracted increasing attention. Superovulation with exogenous hormones may prevent oocytes from establishing an appropriate epigenetic state, and this effect may extend to the methylation programming in preimplantation embryos, as de novo DNA methylation is a function of developmental stage of follicles and oocyte size. Follicle-stimulating hormone (FSH) and human menopausal gonadotropin (hMG) are common gonadotropins used for superovulation, and appropriate concentrations of these gonadotropins might be necessary. However, no systematic study on the effects of DNA methylation alterations in oocytes associated with superovulation with different dosages of FSH/hMG at the single-cell level has yet been reported. In the current study, different dosages of FSH/hMG combined with human chorionic gonadotropin (hCG) were used in female mice to generate experimental groups, while naturally matured oocytes and oocytes superovulated with only hCG were respectively used as controls. Single-cell level DNA methylation sequencing was carried out on all these matured oocytes. RESULTS: In this study, we revealed that the genome-wide methylation pattern and CG methylation level of the maternal imprinting control regions of all mature oocytes were globally conserved and stable. However, methylation alterations associated with superovulation were found at a specific set of loci, and the differentially methylated regions (DMRs) mainly occurred in regions other than promoters. Furthermore, some of the annotated genes in the DMRs were involved in biological processes such as glucose metabolism, nervous system development, cell cycle, cell proliferation, and embryo implantation and were altered in all dosages of FSH/hMG group (for example, Gfod2 and SYF2). Other genes were impaired only after high gonadotropin dosages (for instance, Sox17 and Phactr4). CONCLUSIONS: In conclusion, the current study addressed the effects of superovulation on DNA methylation from the perspective of different dosages of gonadotropins at the single-cell level. We found that the genome-wide DNA methylation landscape was globally preserved irrespective of superovulation or of the kind and dosage of gonadotropins used, whereas the methylation alterations associated with superovulation occurred at a specific set of loci. These observed effects reflect that superovulation recruits oocytes that would not normally be ovulated or that have not undergone complete epigenetic maturation. Our results provide an important reference for the safety assessment of superovulation with different dosages of gonadotropins. However, it should be noted that this study has some limitations, as the sample number and library coverage of analyzed oocytes were relatively low. Future studies with larger sample sizes and high-coverage libraries that examine the effects of superovulation on embryo development and offspring health as well as the underlying mechanisms are still needed.
Assuntos
Gonadotropina Coriônica/farmacologia , Metilação de DNA/efeitos dos fármacos , Epigênese Genética/efeitos dos fármacos , Oócitos/metabolismo , Análise de Sequência de DNA/métodos , Animais , Blastocisto/efeitos dos fármacos , Blastocisto/metabolismo , Gonadotropina Coriônica/administração & dosagem , Gonadotropina Coriônica/metabolismo , Metilação de DNA/genética , Epigênese Genética/genética , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Variantes Farmacogenômicos/genética , Superovulação/efeitos dos fármacos , Superovulação/metabolismoRESUMO
STUDY QUESTION: Could in vitro maturation (IVM) following transvaginal oocyte retrieval during gynaecological surgery (IVM-surgery) be an effective and safe strategy for fertility preservation? SUMMARY ANSWER: IVM-surgery on unstimulated ovaries is a novel option that can be considered for fertility preservation for women requiring gynaecological surgery, but more research is needed to identify appropriate patients who may benefit and to determine the cost-effectiveness of such an approach. WHAT IS KNOWN ALREADY: IVM followed by oocyte/embryo cryopreservation has been useful as a safe reproductive strategy for some infertile women. STUDY DESIGN, SIZE, DURATION: This prospective cohort study comprised 158 consecutive women with polycystic ovary syndrome (PCOS) who underwent laparoscopy or hysteroscopy for other reasons and had concomitant transvaginal oocyte retrieval followed by IVM between 2014 and 2016. PARTICIPANTS/MATERIALS, SETTING, METHODS: A total of 158 women with anovulatory PCOS who underwent IVM-surgery in our infertility centre were recruited for this study. Matured IVM oocytes obtained from these women were either freshly fertilized and subsequently frozen at the blastocyst stage (fresh oocyte group, n = 46) or the oocytes were frozen (frozen oocyte group, n = 112) for fertility preservation followed by later thawing for insemination and cleavage embryo transfer (ET) (n = 33). The following outcomes were then evaluated: embryological data, clinical pregnancy rate, live birth rate (LBR), neonatal outcomes, post-operative complications and post-operative ovarian function. MAIN RESULTS AND THE ROLE OF CHANCE: Among all the women who underwent IVM-surgery, the clinical pregnancy rate and LBR per initiated IVM cycle were 9.5% (15/158) and 6.9% (11/158), respectively. Women (40.6%, 20/33) who underwent the procedure with frozen-thawed oocytes (oocyte survival rate, 83.0%) obtained a high quality of cleaved embryos. In the fresh oocyte group, the clinical pregnancy rate and LBR per ET cycle were 69.2 and 53.8%, respectively. In the frozen oocyte group, the clinical pregnancy rate and LBR per ET cycle were 28.6 and 19.1%, respectively. No adverse neonatal outcomes were recorded. IVM-surgery was not associated with post-operative complications, a longer hospital stay, or impaired ovarian function. LIMITATIONS, REASONS FOR CAUTION: Because of the small sample size and the low utilization rate and cost-effectiveness per retrieval, the present findings should be interpreted with caution, and further studies are needed for the long-term follow-up of live births. WIDER IMPLICATIONS OF THE FINDINGS: This strategy can also help patients with normal ovulation to obtain available oocytes and embryos for cryopreservation and subsequent use. STUDY FUNDING/COMPETING INTEREST(S): This research was supported by the Joint Research Fund for Overseas Natural Science of China (No. 31429004), the National Key Research and Development Program of China (No. 2017YFC1002000, 2017YFC1001504, 2016YFC1000302), the Ministry of Science and Technology of China Grants (No. 2014CB943203), the Chinese Society of Reproductive Medicine Fund (No. 16020400656) and the National Natural Science Foundation of China (No. 81300456). All the authors have nothing to disclose in terms of conflicts of interest. TRIAL REGISTRATION NUMBER: chictr-ONC-17011861.
Assuntos
Preservação da Fertilidade , Infertilidade Feminina , China , Feminino , Humanos , Técnicas de Maturação in Vitro de Oócitos , Infertilidade Feminina/terapia , Recuperação de Oócitos , Oócitos , Gravidez , Taxa de Gravidez , Estudos Prospectivos , Estudos RetrospectivosRESUMO
BACKGROUND: Superovulation treatment had some adverse effects on maturity and development of oocytes. Can superovulation dose of gonadotropins (Gns) affect the transcriptome of granulosa cells and follicular fluid (FF) hormone levels? METHODS: One leading pre-ovulatory follicle per subject was used from three natural-cycle and four Gn-stimulated patients. Granulosa cells and FF samples were collected from the same leading follicle of each patient. RNA was extracted from granulosa cells and subjected to deep sequencing and analysis. Follicle-stimulating hormone (FSH), estradiol (E2), androstenedione (AND), testosterone (T), luteinizing hormone (LH), and progesterone (P4) levels in FF were measured by immunoassays. Student's t test was used for statistical analysis. RESULTS: A total of 715 genes were up-regulated, and 287 genes were down-regulated, in the Gn-stimulated group relative to the control group. Gene Ontology analysis revealed that the down-regulated genes were enriched in cell cycle and meiosis pathways, primarily those associated with follicle or oocyte maturation and quality. On the other hand, the up-regulated genes were enriched in functions related to immunity and cytokine-cytokine receptor interactions. Compared to the follicles of natural cycle, the E2 and LH concentrations were significantly reduced (P < 0.001), the P4 concentration was significantly increased (P = 0.003), and the concentrations of FSH, T and AND had no difference in the follicles of Gn-stimulated cycle. CONCLUSIONS: Cell cycle- and meiosis-associated genes were down-regulated by Gns stimulation, whereas immune- and cytokine-associated genes were up-regulated. Hormone levels were also affected by Gns stimulation. Compared with natural-cycle follicles,putative markers associated with oocyte quality and follicle maturation were significantly different from those in Gn-stimulated follicles. Hormone levels in follicles were compatible with the steroidogenic patterns of granulosa cell, which reflects the follicle maturation and oocyte quality.
Assuntos
Líquido Folicular/metabolismo , Gonadotropinas/farmacologia , Células da Granulosa/metabolismo , Hormônios Hipofisários/metabolismo , Transcriptoma/efeitos dos fármacos , Androstenodiona/metabolismo , Estradiol/metabolismo , Feminino , Fertilização in vitro , Hormônio Foliculoestimulante/metabolismo , Ontologia Genética , Humanos , Hormônio Luteinizante/metabolismo , Transdução de Sinais/genética , Testosterona/metabolismoRESUMO
Gonadotropins have been widely used in human-assisted reproduction and animal science for the past four decades. However, the effects of gonadotropins on oocyte maturation at the molecular and biochemical levels are poorly understood. To determine the effects of gonadotropins (recombinant follicle stimulating hormone and urinary human menopausal gonadotropin) on oocyte maturation, we used the bovine oocyte in vitro maturation model. First, we studied the effects of increasing gonadotropin concentrations on nuclear maturation and mitochondrial function in oocytes. Gonadotropins at concentrations of 0.075 and 0.75 IU/ml improved nuclear maturation and increased inner mitochondrial membrane potential and ATP levels; however, there were no beneficial effects at concentrations of 7.5 and 75 IU/ml. Second, we studied the effects of increasing gonadotropin concentrations on the status of methylation in matured (MII) oocytes. Aberrant methylation and demethylation of H19, SNRPN, and PEG3 genes were observed in MII oocytes at all concentrations except 0.075 IU/ml. The expression of genes that function in spindle formation, cell cycle control, and methylation was also downregulated by high gonadotropin concentrations. In conclusion, we established the optimal gonadotropin concentration (i.e., 0.075 IU/ml) to be used for bovine oocyte in vitro maturation studies. These results may provide a guide for clinical stimulation protocols and help to reduce the risks associated with gonadotropin administration during in vitro fertilization treatment.
Assuntos
Bovinos/fisiologia , Gonadotropinas/farmacologia , Técnicas de Maturação in Vitro de Oócitos/veterinária , Oócitos/citologia , Oócitos/fisiologia , Animais , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/fisiologia , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/fisiologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Coloração e RotulagemRESUMO
It is well known that maternal ageing not only causes increased spontaneous abortion and reduced fertility, but it is also a high genetic disease risk. Although assisted reproductive technologies (ARTs) have been widely used to treat infertility, the overall success is still low. The main reasons for age-related changes include reduced follicle number, compromised oocyte quality especially aneuploidy, altered reproductive endocrinology, and increased reproductive tract defect. Various approaches for improving or treating infertility in aged women including controlled ovarian hyperstimulation with intrauterine insemination (IUI), IVF/ICSI-ET, ovarian reserve testing, preimplantation genetic diagnosis and screening (PGD/PGS), oocyte selection and donation, oocyte and ovary tissue cryopreservation before ageing, miscarriage prevention, and caloric restriction are summarized in this review. Future potential reproductive techniques for infertile older women including oocyte and zygote micromanipulations, derivation of oocytes from germ stem cells, ES cells, and iPS cells, as well as through bone marrow transplantation are discussed.
Assuntos
Envelhecimento/fisiologia , Infertilidade Feminina/terapia , Técnicas Reprodutivas , Transplante de Medula Óssea , Feminino , Humanos , Idade MaternaRESUMO
STUDY QUESTION: Do different concentrations of FSH in the assisted reproductive technology (ART) procedure in vitro or in vivo affect the developmental competence of oocytes, the embryos and the offspring conceived from these embryos? SUMMARY ANSWER: Improper FSH treatment (200 IU/l in vitro, 10 IU/ml in vivo and 200 IU/ml in vivo) impairs the development competence of oocyte and embryo, but does not influence offspring physiology and behavior. WHAT IS KNOWN ALREADY: Exogenous FSH has been widely used in the field of ART. However, the effects of different concentrations of FSH on the developmental competence of oocytes, embryos and the offspring conceived from these embryos, are still unknown. STUDY DESIGN, SIZE, DURATION: In a prospective study, a total of 45 mice at 8-10 weeks of age were primed in vivo with different dosages of FSH (9 mice in the 10 IU/ml, 10 mice in the 50 IU/ml, 10 mice in the 100 IU/ml and 16 mice in the 200 IU/ml groups). Fresh MII oocytes were retrieved from ovaries: this was designated as in vivo group. Thirty six mice at 8-10 weeks of age were sacrificed by cervical dislocation to obtain ovaries without FSH treatment (9 mice in the 0 IU/l, 9 mice in the 50 IU/l, 8 mice in the 100 IU/l and 10 mice in the 200 IU/l groups), and then the immature oocytes were collected from these ovaries and cultured in vitro matured medium supplemented with 0, 50, 100 and 200 IU/l FSH: this was designated as in vitro group. MATERIALS, SETTING, METHODS: Spindle assembly of matured MII oocytes was stained via an immunofluorescence method and the oocytes ratio of normal spindle was analyzed. The developmental competence of the resulting fertilized embryos in the pre- and post-implantation stages was examined in in vitro and in vivo groups. Furthermore, physiological index, including reproductive potential and body weight, of the offspring was investigated by mating experiments and behavior index, including learning, memory, probing and intelligence, was tested by Morris water maze in in vitro and in vivo groups. MAIN RESULTS AND THE ROLE OF CHANCE: In the in vitro groups, the oocyte maturation competence, normal spindle assembly, blastocyst formation and implantation, as well as viable pup production were all impaired in the group treated with 200 IU/l FSH (P < 0.05). No differences were observed among the other three groups (P > 0.05). In the in vivo groups, 10 IU/ml FSH but not 200 IU/ml treatment influenced blastocyst formation and viable pup production (P < 0.05), although the high proportion of spindle assembly abnormality was only observed in the 200 IU/ml FSH treatment group (P < 0.05). Furthermore, there were no significant differences in terms of physiological index (reproductive potential and body weight) and behavior index (learning, memory, probing and intelligence) in offspring from in vitro and in vivo groups (P > 0.05). LIMITATIONS, REASONS FOR CAUTION: The mouse model was used in this study. The results of the mouse follicle growth and oocyte development in responding to different concentrations of FSH are not 100% transferable to human, because of the physiological differences between mouse and human. WIDER IMPLICATIONS OF THE FINDINGS: The findings indicated that FSH application in the field of ART is safe to the resulted offspring, but it should be more carefully used for each women in ART cycles because the inappropriate FSH concentration would decrease the oocyte developmental competence. STUDY FUNDING/COMPETING INTEREST(S): This work was partially supported by the Ministry of Science and Technology of China Grants (973 program; 2011CB944504), the Program for Changjiang Scholars and Innovative Research Team in University of Ministry of Education of China (30825038), the National Natural Science Funds for Young Scholar (31000661) and by the Joint Research Fund for Overseas, Hong Kong and Marco Scholars (31128013/C120205). None of the authors has any conflicts of interest.
Assuntos
Fertilização/fisiologia , Hormônio Foliculoestimulante/metabolismo , Oócitos/fisiologia , Técnicas de Reprodução Assistida , Animais , Comportamento Animal , Blastocisto/citologia , Meios de Cultura/química , Técnicas de Cultura Embrionária , Feminino , Hormônio Foliculoestimulante/farmacologia , Humanos , Testes de Inteligência , Masculino , Memória , Camundongos , Camundongos Endogâmicos ICR , Microscopia de Fluorescência , Oócitos/citologia , Folículo Ovariano/fisiologia , Injeções de Esperma Intracitoplásmicas , Espermatozoides/metabolismo , Espermatozoides/patologiaRESUMO
BACKGROUND: It remains almost a helpless situation for the recurrent implantation failure and pregnancy loss caused by endometrial injury at present. The purpose of this study was to develop a rabbit model of endometrial mechanical injury that could provide a research platform for this difficult clinical predicament. METHODS: Three experiments were conducted. Experiment 1: Curettages in both uterus horns and copper wire inserting after curettage (double-injury) in one horn. The histological changes were monitored at 0, 24, 48, 72 hours, as well as in 1 and 2 weeks after operation. Experiment 2: Direct copper wire inserting in one horn and double-injury in other horn. The wires in both horns were removed after 2 weeks. The histological changes were recorded at 0, 1 and 2 weeks after wire removal. Experiment 3: Double-injury procedure in one horn was performed and wire was removed after 2 weeks; another horn was remained normal to serve as control. Histological changes were recorded, tissue areas were measured, and proliferation indices (PIs, %) were calculated at 1, 2, 4 and 8 weeks after wire removal, respectively. RESULTS: The experiments revealed that the injured endometrium by simple curettage or copper wire could be fully repaired. While the endometrial regeneration was severely impaired by double-injury, both areas of endometrium and uterine cavity decreased (P < 0.05); both PIs of glandular epithelial and stromal cells increased and reached maximum at 4 weeks (P < 0.05), but returned by 8 weeks. CONCLUSION: This study demonstrated that a rabbit model of endometrial injury could be effectively established through a double-injury procedure of curettage and copper wire with comparable clinical index.
Assuntos
Cobre/efeitos adversos , Curetagem/efeitos adversos , Modelos Animais de Doenças , Endométrio/lesões , Animais , Feminino , Imuno-Histoquímica , CoelhosRESUMO
The objective of the present study was to evaluate the developmental potential and clinical application value of metaphase I (MI) oocytes obtained from stimulated intracytoplasmic sperm injection (ICSI) cycles. ICSI was performed on MI oocytes immediately after denudation (Group A), or on in vitro-matured (IVM) oocytes following culture; oocytes in culture were further divided into two groups, being cultured for either 3-5 h (Group B) or 24-28 h (Group C). Metaphase II oocytes from the same cycle(s) isolated for ICSI served as the control group (Group D). The rates of normal fertilisation, cleavage and high-quality embryos were compared among the four groups. High-quality embryos were transferred whenever possible, and pregnancy rates were evaluated. Results showed that normal fertilisation rates for Groups B, C and D were significantly higher than that of Group A (68.6%, 57.8%, 74.5% and 30.1%, respectively; P<0.01). The rate of high-quality embryos in Group B was comparable with Group D; the rate for Group C was significantly lower than that of the other groups (P<0.05). Two clinical pregnancies were achieved after transfer of embryos from IVM oocytes. In vitro maturation of MI oocytes for a short period of time may increase the number of available embryos; however, overnight in vitro culture of MI oocytes did not improve results.
Assuntos
Metáfase/fisiologia , Oócitos/fisiologia , Injeções de Esperma Intracitoplásmicas/métodos , Adulto , Distribuição de Qui-Quadrado , Transferência Embrionária/métodos , Desenvolvimento Embrionário/fisiologia , Feminino , Humanos , Masculino , Oócitos/citologia , Gravidez , Fatores de TempoRESUMO
This paper aims to determine the possible role of estrogen receptor-ß (ERß) gene RsaI polymorphism on sperm fertility and early embryonic development in humans. Three groups of Chinese men were recruited: in vitro fertilization (IVF) group, including 374 couples who underwent conventional IVF; intracytoplasmic sperm injection (ICSI) group, including 294 couples who underwent an ICSI procedure using ejaculated sperm; and azoospermic group, consisting of 197 couples who underwent ICSI using either testis or epididymis sperm. RsaI polymorphism in the ERß gene was detected by polymerase chain reaction (PCR)-restriction fragment length polymorphism technique; fertilization and high-quality embryo rates were evaluated for each group. In each group, no significant differences were found in the overall rates of fertilization and high-quality embryos among GG, AG and AA genotypes. However, the proportion of cycles possessing a satisfactory high-quality embryo rate with the AA genotype was significantly lower than that in the wild-type GG genotype from each group. These results demonstrated that sperm possessing the ERß RsaI A genotype may have reduced fertilization ability and decreased early embryonic developmental potential, which could directly or indirectly contribute to the low fertilization rate and early embryonic developmental arrest in some cases.
Assuntos
Desenvolvimento Embrionário/genética , Desenvolvimento Embrionário/fisiologia , Receptor beta de Estrogênio/genética , Receptor beta de Estrogênio/fisiologia , Fertilidade/genética , Fertilidade/fisiologia , Infertilidade Masculina/genética , Infertilidade Masculina/fisiopatologia , Povo Asiático/genética , Azoospermia/genética , Azoospermia/fisiopatologia , Azoospermia/terapia , Sequência de Bases , China , Primers do DNA/genética , Feminino , Fertilização in vitro , Frequência do Gene , Genótipo , Humanos , Infertilidade Masculina/terapia , Masculino , Polimorfismo de Fragmento de Restrição , Gravidez , Injeções de Esperma IntracitoplásmicasRESUMO
OBJECTIVE: To establish a novel scoring system for human immature oocytes and estimate its prognostic effects on the in vitro maturation (IVM) and developmental potential of human immature oocytes. DESIGN: Prospective study. SETTING: Hospital-based IVF center. PATIENT(S): Patients with polycystic ovary syndrome (PCOS) undergoing unstimulated cycles of oocyte retrieval. INTERVENTION(S): Immature (germinal vesicle and metaphase I) oocytes were collected from PCOS patients and were evaluated based on morphologic characters. After IVM, the maturation rate, fertilization rate, and cleavage rate were evaluated. The quality of the embryos and the apoptosis of the granulosa cells (GCs) were analyzed as well. MAIN OUTCOME MEASURE(S): Morphologic characters of immature oocytes and maturation rate, fertilization rate, cleavage rate, high-quality embryo formation rate, and the apoptosis of the GCs after IVM. RESULT(S): The maturation rate, fertilization rate, cleavage rate, and high-quality embryo formation rate of both MI and GV oocytes reduced with the decrease of the score. There were no differences in maturation, fertilization, and cleavage rates between the MI and GV phase. The apoptosis rate of GCs of grade 1' and 0' oocytes was obvious higher than those of grade 3-4'. CONCLUSION(S): Evaluating cumulus layer around the oocyte was a simple but useful method, which played an important role in terms of yielding good prognostic parameters in an embryology laboratory. The scoring system of immature oocytes can correctly reflect the development potential of human oocytes.
Assuntos
Técnicas de Diagnóstico Obstétrico e Ginecológico , Infertilidade Feminina/diagnóstico , Oócitos/patologia , Síndrome do Ovário Policístico/patologia , Apoptose/fisiologia , Diferenciação Celular/fisiologia , Células Cultivadas , Fase de Clivagem do Zigoto/patologia , Fase de Clivagem do Zigoto/fisiologia , Técnicas de Cultura Embrionária , Desenvolvimento Embrionário/fisiologia , Feminino , Fertilização in vitro , Células da Granulosa/patologia , Células da Granulosa/fisiologia , Humanos , Infertilidade Feminina/etiologia , Infertilidade Feminina/patologia , Infertilidade Feminina/fisiopatologia , Oócitos/fisiologia , Oogênese/fisiologia , Indução da Ovulação , Síndrome do Ovário Policístico/complicações , Síndrome do Ovário Policístico/diagnóstico , Síndrome do Ovário Policístico/fisiopatologia , Controle de Qualidade , Projetos de PesquisaRESUMO
AIM: To determine the possible roles of the t-complex testis expressed gene 5 (Tctex5) on sperm functions, the full-length sequence of mRNA was studied and compared in the testis between the normal wild-type and the sterile t-haplotype mutant mice. METHODS: We applied rapid amplification of cDNA ends, Northern blot and reverse transcription polymerase chain reaction to analyze the full length of Tctex5 mRNAs isolated from testes of the wild-type and the t-haplotype mice. Reverse transcription polymerase chain reaction was used to semi-quantitatively compare expression of Tctex5 transcripts in the 16 tissues and 9.5 day stage embryos in the wild-type mice. E-translation was applied to estimate the amino acid sequences. RESULTS: One long and one short transcript of Tctex5 mRNA were discovered in mouse testis of wild-type (Tctex5(long-+) and Tctex5(short-+)) and t-haplotype (Tctex5(long-t) and Tctex5(short-t)) mice, respectively. Being enhanced only in the testis, Tctex5(long-t) had 17 point mutations and one 15-bp-deletion in the exon 1 region, comparing with the Tctex5(long-+), whereas the Tctex5(short-t) was similar to the Tctex5(short-+). The short isoforms of Tctex5 mRNAs in the two models encoded exactly the same peptides, but the long isoforms did not. The estimated peptide encoded by Tctex5(long-t) had significant mutations on putative sites of phosphorylation and PP1 binding. CONCLUSION: We established that mutations that occur in the Tctex5 long transcript of the t-haplotype mice are important for normal sperm function, whereas the short transcript of Tctex5 might have a conserved function among different tissues.
Assuntos
Proteínas Associadas aos Microtúbulos/química , Proteínas Associadas aos Microtúbulos/genética , Proteínas Nucleares/química , Proteínas Nucleares/genética , Espermatozoides/metabolismo , Testículo/metabolismo , Animais , Expressão Gênica , Haplótipos , Infertilidade Masculina , Masculino , Camundongos , Mutação , Proteína Fosfatase 1 , Análise de Sequência de Proteína , Ubiquitina-Proteína Ligases , Região do Complexo-t do GenomaRESUMO
OBJECTIVE: To assess the potential effects of in vitro maturation (IVM) of human oocytes on the meiotic spindle and associated chromosome configuration. DESIGN: Prospective study. SETTING: Hospital-based IVF center. PATIENT(S): Patients with polycystic ovary syndrome (PCOS) undergoing unstimulated and stimulated cycles of oocyte retrieval. INTERVENTION(S): Immature (germinal vesicle and metaphase I) and mature (metaphase II) oocytes were collected from PCOS patients. The meiotic spindle and chromosome configurations in oocytes matured in vitro and in vivo were studied by confocal microscopy, with fluorescent labeling techniques for visualization of both microtubules and chromatin. MAIN OUTCOME MEASURE(S): Meiotic spindle and associated chromosome configurations. RESULT(S): Oocytes can develop to the metaphase II stage after IVM. Confocal microscopic observations revealed that the oocytes matured in vitro had a higher frequency of abnormal meiotic spindle and chromosomal alignment morphology than in vivo-matured oocytes. These abnormalities included a partial or total disorganization of the meiotic spindle microtubules. Abnormal chromosome organization included dispersal of chromosomes or chromosomes with an aberrant, less-condensed appearance. The proportions of abnormality in spindle and chromosome configurations in oocytes matured in vitro were 43.7% and 33.3%, respectively, which was significantly higher than in those oocytes matured in vivo (13.6% and 9.1%). CONCLUSION(S): In vitro maturation can have deleterious effects on the organization of the meiotic spindle and chromosome alignment of human oocytes. This result suggests one possible explanation for the reduced developmental potential of oocytes matured in vitro compared with those matured in vivo. This is likely a contributing factor to the overall lower clinical outcomes observed after IVM and ET.