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1.
Parasit Vectors ; 12(1): 205, 2019 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-31060579

RESUMO

BACKGROUND: Vitellogenin (Vg), a key molecule for oocyte development synthesized in the fat body during blood-feeding, is released into the hemolymph and then taken into the oocytes via Vg receptor (VgR) in ticks. Previously, we showed that VgR mRNA is expressed in the ovary at the adult stage of parthenogenetic Haemaphysalis longicornis ticks and its expression increases after blood-feeding. However, intracellular localization of VgR mRNA and protein at each developmental stage of oocytes during oogenesis remains largely unclear. METHODS: mRNA and protein expression profiles of H. longicornis VgR (HlVgR) in the oocytes from the unfed to oviposition periods were analyzed by real-time PCR, in situ hybridization, and immunostaining. To elucidate the timing of the onset of Vg uptake, RNA interference (RNAi)-mediated gene silencing of HlVgR was performed. RESULTS: In situ hybridization revealed that HlVgR mRNA was detected in the cytoplasm of stage I-III oocytes, and weaker positive signals for HlVgR mRNA were found in the cell periphery of stage IV and V oocytes. Likewise, HlVgR protein was detected by immunostaining in the cytoplasm of stage I-III oocytes and in the cell periphery of stage IV and V oocytes. Each developmental stage of the oocytes showed distinct patterns of mRNA and protein expression of HlVgR. Moreover, RNAi of HlVgR caused delayed or arrested development in the oocytes. The ovaries of control ticks showed all developmental stages of oocytes, whereas stage I-III oocytes were found in the ovaries of HlVgR-RNAi ticks at 5 days after engorgement. CONCLUSIONS: These results suggest that active uptake of Vg is required for development from stage III to stage IV during oogenesis. Our data clearly revealed an apparent shift in the intracellular localization of VgR for both mRNA and protein level in oocytes during oogenesis.


Assuntos
Proteínas do Ovo/metabolismo , Ixodidae/metabolismo , Oogênese/fisiologia , RNA Mensageiro/metabolismo , Receptores de Superfície Celular/metabolismo , Vitelogeninas/metabolismo , Animais , Proteínas do Ovo/genética , Feminino , Ixodidae/genética , Ixodidae/crescimento & desenvolvimento , Camundongos , Camundongos Endogâmicos BALB C , Oócitos/crescimento & desenvolvimento , Oócitos/metabolismo , Oogênese/genética , Coelhos , Reação em Cadeia da Polimerase em Tempo Real , Receptores de Superfície Celular/genética , Transcriptoma
2.
Arthropod Struct Dev ; 50: 53-63, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31004762

RESUMO

In this study, we describe the female reproductive system organization and oogenesis in the eutardigrade Thulinius ruffoi. Light, confocal and electron microscopy was used in this study. During oogenesis, three phases can be distinguished: previtellogenesis, vitellogenesis, and choriogenesis. Germ-line cells form cell clusters in which the cells are connected by intercellular (cytoplasmic) bridges. These structures are crucial for delivering the yolk materials, macromolecules, ribosomes, and organelles to the developing oocyte. Vitellogenesis is of a mixed type. Autosynthesis and heterosynthesis of the yolk material occur. Yolk precursors that have been synthesized outside the ovary are delivered to the oocyte via endocytosis. We also present data on cortical granules, and moreover, we describe the cortical reaction in tardigrades, possibly for the first time.


Assuntos
Tardígrados/anatomia & histologia , Tardígrados/fisiologia , Animais , Feminino , Genitália Feminina/anatomia & histologia , Genitália Feminina/ultraestrutura , Microscopia Confocal , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Oogênese/fisiologia , Tardígrados/ultraestrutura
3.
Gene ; 701: 104-112, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-30905810

RESUMO

PIWI family member piwil1, which associates with Piwi-interacting RNA (piRNA), is responsible in regulation of germ cell differentiation and maintenance of reproductive stem cells. In this study, we analyzed the piwil1 gene in Paralichthys olivaceus. Bioinformatics analysis and structure prediction showed that piwil1 had the conserved domains: PAZ domain and PIWI domain. Expression analysis during embryonic development implied that piwil1 gene was maternally inherited. The tissue distribution showed a sexually dimorphic gene expression pattern, with higher expression level in testis than ovary. In situ hybridization results demonstrated that piwil1 was predominantly distributed in oogonia, oocytes, sertoli cells and spermatocytes. A CpG island was predicted in the 5'-flanking region of piwil1 gene, and its methylation levels showed significant disparity between males and females, indicating that the sexually dimorphic expression of piwil1 gene might be regulated by methylation. Furthermore, we explored the distinct roles of human chorionic gonadotropin and 17α-methyltestosterone in regulating the expression of piwil1, and found that piwil1 was interacting with the HPG axis hormones. These results indicated that piwil1 might play a crucial role in gonadal development and gametogenesis in Paralichthys olivaceus.


Assuntos
Proteínas Argonauta/biossíntese , Proteínas de Peixes/biossíntese , Linguado/crescimento & desenvolvimento , Regulação da Expressão Gênica , Oogênese/fisiologia , Espermatogênese/fisiologia , Animais , Proteínas Argonauta/genética , Feminino , Proteínas de Peixes/genética , Linguado/genética , Masculino , Oócitos/citologia , Oócitos/metabolismo , Oogônios/citologia , Oogônios/metabolismo , Células de Sertoli/citologia , Células de Sertoli/metabolismo
4.
J Reprod Dev ; 65(2): 183-190, 2019 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-30745496

RESUMO

We examined whether the use of in vivo-matured oocytes, collected by ovum pick-up (OPU) from superstimulated Japanese Black cows, can improve the productivity and quality of in vitro produced embryos. The cows were superstimulated by treatment with progesterone, GnRH, FSH and prostaglandin F2α according to a standardized protocol. The resulting in vivo-matured oocytes were collected by OPU and used subsequently for the other experiments. The immature oocytes from cows in the non-stimulated group were collected by OPU and then subjected to maturation in vitro. We found that the rate of normally distributed cortical granules of the matured oocyte cytoplasm in the superstimulated group was significantly higher than that in the non-stimulated group. The normal cleavage rate (i.e., production of embryos with two equal blastomeres without fragmentation) and freezable blastocyst rate were significantly higher in the superstimulated group than in the non-stimulated group. Among the transferable blastocysts, the ratio of embryos from normal cleavage was also significantly higher in the superstimulated group than in the non-stimulated group. For in vivo-matured oocytes, it was observed that the pregnancy rates were significantly higher when normally cleaved embryos were used for transfer. Taken together, these results suggest that high-quality embryos with respect to developmental kinetics can be efficiently produced with the use of in vivo-matured oocytes collected by OPU from superstimulated Japanese Black cows.


Assuntos
Bovinos , Embrião de Mamíferos/citologia , Fertilização In Vitro/métodos , Técnicas de Maturação in Vitro de Oócitos , Recuperação de Oócitos , Oócitos/fisiologia , Indução da Ovulação , Animais , Transferência Embrionária/veterinária , Desenvolvimento Embrionário/fisiologia , Feminino , Fertilização In Vitro/veterinária , Técnicas de Maturação in Vitro de Oócitos/métodos , Técnicas de Maturação in Vitro de Oócitos/veterinária , Masculino , Recuperação de Oócitos/veterinária , Oócitos/citologia , Oogênese/fisiologia , Indução da Ovulação/veterinária , Gravidez , Taxa de Gravidez , Resultado do Tratamento
5.
J Reprod Dev ; 65(2): 113-120, 2019 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-30606957

RESUMO

This study aimed to investigate the effect of resveratrol supplementation in maturation medium on the developmental ability and bioenergetic\oxidative status of prepubertal goat oocytes selected by brilliant cresyl blue (BCB). Oocytes collected from slaughterhouse-derived ovaries were selected by 13 µM BCB staining and classified as grown BCB+ and growing BCB- oocytes. All oocytes were matured in vitro in our conventional maturation medium and supplemented with 1 µM (BCB+R and BCB-R) and without (Control groups: BCB+C and BCB-C) resveratrol. After 24 h, IVM-oocytes were fertilized with fresh semen and presumptive zygotes were in vitro cultured for 8 days. Oocytes were assessed for blastocyst development and quality, mitochondrial activity and distribution, and levels of GSH, ROS, and ATP. BCB+R (28.3%) oocytes matured with resveratrol presented significantly higher blastocyst development than BCB+C (13.0%) and BCB- groups (BCB-R: 8.3% and BCB-C: 4.7%). Resveratrol improved blastocyst development of BCB-R oocytes at the same rate as BCB+C oocytes. No differences were observed in blastocyst quality among groups. GSH levels were significantly higher in resveratrol groups (BCB+R: 36554.6; BCB-R: 34946.7 pixels/oocyte) than in control groups (BCB+C: 27624.0; BCB-C: 27655.4 pixels/oocyte). No differences were found in mitochondrial activity, ROS level, and ATP content among the groups. Resveratrol-treated oocytes had a higher proportion of clustered active mitochondria in both BCB groups (BCB+R: 73.07%; BCB-R: 79.16%) than control groups (BCB+C: 19.35%; BCB-C: 40%). In conclusion, resveratrol increased blastocyst production from oocytes of prepubertal goats, particularly in better quality oocytes (BCB+).


Assuntos
Desenvolvimento Embrionário/efeitos dos fármacos , Técnicas de Maturação in Vitro de Oócitos , Oócitos/citologia , Oócitos/efeitos dos fármacos , Resveratrol/farmacologia , Maturidade Sexual/fisiologia , Animais , Blastocisto/efeitos dos fármacos , Blastocisto/fisiologia , Separação Celular/métodos , Células Cultivadas , Corantes/química , Embrião de Mamíferos , Feminino , Fertilização In Vitro/veterinária , Cabras , Técnicas de Maturação in Vitro de Oócitos/métodos , Técnicas de Maturação in Vitro de Oócitos/veterinária , Masculino , Oócitos/fisiologia , Oogênese/efeitos dos fármacos , Oogênese/fisiologia , Oxazinas/química , Coloração e Rotulagem/métodos
6.
Sci Rep ; 8(1): 17737, 2018 12 10.
Artigo em Inglês | MEDLINE | ID: mdl-30531803

RESUMO

RING-between-RING (RBR) E3 ubiquitin ligases are implicated in various developmental processes, and mutations in genes encoding RBR proteins HHARI/ARIH1 and Parkin are associated with human diseases. Here we show by phylogenetic analysis that the ARI1 family has undergone a dramatic expansion within the Caenorhabditis clade in recent history, a characteristic shared by some genes involved in germline development. We then examined the effects of deleting all ARI1 family members in the nematode Caenorhabditis elegans, which to our knowledge represents the first complete knockout of ARI1 function in a metazoan. Hermaphrodites that lacked or had strongly reduced ARI1 activity had low fecundity and were partially defective in initiation of oocyte differentiation. We provide evidence that the C. elegans ARI1s likely function downstream or in parallel to FBF-1 and FBF-2, two closely related RNA-binding proteins that are required for the switch from spermatogenesis to oogenesis during late larval development. Previous studies have shown that the E2 enzymes UBC-18/UBCH7 and UBC-3/CDC34 can functionally collaborate with ARI1 family members. Our data indicated that UBC-18, but not UBC-3, specifically cooperates with the ARI1s in germline development. These findings provide new insights into the functions of RING-between-RING proteins and Ariadne E3s during development.


Assuntos
Proteínas de Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/crescimento & desenvolvimento , Caenorhabditis elegans/metabolismo , Células Germinativas/crescimento & desenvolvimento , Células Germinativas/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitina/metabolismo , Animais , Diferenciação Celular/fisiologia , Oócitos/metabolismo , Oogênese/fisiologia , Filogenia , Proteínas de Ligação a RNA/metabolismo , Espermatogênese/fisiologia , Enzimas de Conjugação de Ubiquitina/metabolismo
7.
PLoS One ; 13(12): e0208760, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30532263

RESUMO

The present study evaluated the effect of supplementing in vitro culture medium with J. insularis compared to FSH on isolated secondary follicles and in vitro maturation of oocytes from those follicles. Secondary follicles were isolated from sheep ovaries and individually cultured for 18 days in α-MEM+ (Control), α-MEM+ supplemented with 100 ng/mL recombinant bovine follicle stimulating hormone (FSH) or with 0.3, 1.25, or 2.5 mg/mL of J. insularis extract (JI0.3, JI1.25, and JI2.5, respectively). Culture medium collected every 2 days was used to measure ROS levels. At the end of the culture period, cumulus oocytes complex (COCs) were collected and matured in vitro. Follicular walls were used for mRNA quantitation. JI0.3 led to a higher (P < 0.05) percentages of intact follicles than other groups after 18 days of culture. While follicular diameter remained unchanged from Day 6 onwards with JI0.3 and FSH, percentages of antral cavity formation were higher (P < 0.05) with JI0.3 at Day 6 than in all other treatments. No differences were observed between controls and treatment groups regarding ROS levels and mRNA expression of genes. Viability of resulting oocytes was higher (P < 0.05) in JI0.3 compared to FSH. Interestingly, in control experiment, supplementation of maturation medium with JI0.3 led to higher (P < 0.05) percentages of metaphase II compared to controls. Although more validations will be needed, it seems that this natural extract could be used as a cheap and easily available alternative to commercial FSH.


Assuntos
Meios de Cultura , Hormônio Foliculoestimulante/administração & dosagem , Técnicas de Maturação in Vitro de Oócitos , Adhatoda , Extratos Vegetais/administração & dosagem , Animais , Relação Dose-Resposta a Droga , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Adhatoda/química , Oócitos/efeitos dos fármacos , Oócitos/crescimento & desenvolvimento , Oócitos/metabolismo , Oogênese/efeitos dos fármacos , Oogênese/fisiologia , Folículo Ovariano/efeitos dos fármacos , Folículo Ovariano/crescimento & desenvolvimento , Folículo Ovariano/metabolismo , Compostos Fitoquímicos/administração & dosagem , Compostos Fitoquímicos/química , Extratos Vegetais/química , Folhas de Planta/química , RNA Mensageiro/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Ovinos
8.
Nat Commun ; 9(1): 5318, 2018 12 14.
Artigo em Inglês | MEDLINE | ID: mdl-30552320

RESUMO

Oocyte-specific miRNA function remains unclear in mice and worms because loss of Dgcr8 and Dicer from mouse and worm oocytes, respectively, does not yield oogenic defects. These data lead to several models: (a) miRNAs are not generated in oocytes; (b) miRNAs are generated but do not perform an oogenic function; (c) functional oocyte miRNAs are generated in a manner independent of these enzymes. Here, we test these models using a combination of genomic, expression and functional analyses on the C. elegans germline. We identify a repertoire of at least twenty-three miRNAs that accumulate in four spatial domains in oocytes. Genetic tests demonstrate that oocyte-expressed miRNAs regulate key oogenic processes within their respective expression domains. Unexpectedly, we find that over half of the oocyte-expressed miRNAs are generated through an unknown Drosha independent mechanism. Thus, a functional miRNA repertoire generated via Drosha dependent and independent pathways regulates C. elegans oocyte development.


Assuntos
Caenorhabditis elegans/genética , Genômica , MicroRNAs/genética , MicroRNAs/metabolismo , Oócitos/crescimento & desenvolvimento , Oócitos/metabolismo , Oogênese/fisiologia , Animais , Caenorhabditis elegans/embriologia , Caenorhabditis elegans/crescimento & desenvolvimento , Proteínas de Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/metabolismo , Fertilidade/genética , Fertilidade/fisiologia , Células Germinativas , Hibridização In Situ , Meiose/fisiologia , Oócitos/citologia , Interferência de RNA , Ribonuclease III/genética , Ribonuclease III/metabolismo
9.
Dev Cell ; 46(3): 285-301.e9, 2018 08 06.
Artigo em Inglês | MEDLINE | ID: mdl-30086300

RESUMO

Phase separation represents an important form of subcellular compartmentalization. However, relatively little is known about how the formation or disassembly of such compartments is regulated. In zebrafish, the Balbiani body (Bb) and the germ plasm (Gp) are intimately linked phase-separated structures essential for germ cell specification and home to many germ cell-specific mRNAs and proteins. Throughout development, these structures occur as a single large aggregate (Bb), which disperses throughout oogenesis and upon fertilization accumulates again into relatively large assemblies (Gp). Formation of the Bb requires Bucky ball (Buc), a protein with prion-like properties. We found that the multi-tudor domain-containing protein Tdrd6a interacts with Buc, affecting its mobility and aggregation properties. Importantly, lack of this regulatory interaction leads to significant defects in germ cell development. Our work presents insights into how prion-like protein aggregations can be regulated and highlights the biological relevance of such regulatory events.


Assuntos
Células Germinativas/metabolismo , Oócitos/metabolismo , Oogênese/fisiologia , Proteínas de Peixe-Zebra/metabolismo , Animais , Citoplasma/metabolismo , Organelas/metabolismo , RNA Mensageiro/metabolismo , Peixe-Zebra
10.
Tissue Cell ; 53: 37-43, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30060825

RESUMO

The morphological and cytological changes of oogenesis and ovarian development were described in the freshwater crab Sinopotamon henanense through macroscopic and microscopic examinations. Serial histological dissections of the ovaries demonstrated that oocyte development was asynchronous. Oogenesis was divided into four distinct stages including six phases: oogonium stage, the first phase (OI) and the second phase (OII) comprising the previtellogenic stage, the third phase (OIII), the fourth phase (OIV) and the fifth phase (OV), comprising the vitellogenic stage and the sixth phase representing the mature stage. Furthermore, examining and analyzing the gonadosomatic indices showed that the developmental cycle of the ovary was closely related to season, and indicated that the breeding season of S. henanense was between May and June. Ovarian development was classified into six stages: proliferation stage, small growth stage, large growth stage, pre-maturation stage, mature stage and spawning stage. Ovaries varied in size and color during each developmental stage, which were closely related to the status and proportions of oogonia and primary oocytes. Although there were cases that oocytes at two or more phases were present at each stage, ovary developmental stages were substantially different. These results provide an important base for studies of the regulatory mechanisms of oogenesis in this compared to other brachyuran species, and will be useful for the aquaculture of S. henanense and related species.


Assuntos
Braquiúros , Oogênese/fisiologia , Ovário , Animais , Braquiúros/citologia , Braquiúros/fisiologia , Feminino , Ovário/citologia , Ovário/fisiologia
11.
Dev Biol ; 441(1): 52-66, 2018 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-29886128

RESUMO

CDK11, a member of the cyclin-dependent kinase family, has been implicated in a diverse array of functions including transcription, RNA processing, sister chromatid cohesion, spindle assembly, centriole duplication and apoptosis. Despite its involvement in many essential functions, little is known about the requirements for CDK11 and its partner Cyclin L in a developing multicellular organism. Here we investigate the function of CDK11 and Cyclin L during development of the nematode Caenorhabditis elegans. Worms express two CDK11 proteins encoded by distinct loci: CDK-11.1 is essential for normal male and female fertility and is broadly expressed in the nuclei of somatic and germ line cells, while CDK-11.2 is nonessential and is enriched in hermaphrodite germ line nuclei beginning in mid pachytene. Hermaphrodites lacking CDK-11.1 develop normally but possess fewer mature sperm and oocytes and do not fully activate the RAS-ERK pathway that is required for oocyte production in response to environmental cues. Most of the sperm and eggs that are produced in cdk-11.1 null animals appear to complete development normally but fail to engage in sperm-oocyte signaling suggesting that CDK-11.1 is needed at multiple points in gametogenesis. Finally, we find that CDK-11.1 and CDK-11.2 function redundantly during embryonic and postembryonic development and likely do so in association with Cyclin L. Our results thus define multiple requirements for CDK-11-Cyclin L during animal development.


Assuntos
Proteínas de Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/embriologia , Quinases Ciclina-Dependentes/metabolismo , Ciclinas/metabolismo , Oogênese/fisiologia , Espermatogênese/fisiologia , Animais , Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/genética , Quinases Ciclina-Dependentes/genética , Ciclinas/genética , Feminino , Fertilidade/fisiologia , Masculino
12.
Dev Biol ; 440(1): 31-39, 2018 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-29729259

RESUMO

Tissue-specific stem cells are tied to the nutritional and physiological environment of adult organisms. Adipocytes have key endocrine and nutrient-sensing roles and have emerged as major players in relaying dietary information to regulate other organs. For example, previous studies in Drosophila melanogaster revealed that amino acid sensing as well as diet-dependent metabolic pathways function in adipocytes to influence the maintenance of female germline stem cells (GSCs). How nutrient-sensing pathways acting within adipocytes influence adult stem cell lineages, however, is just beginning to be elucidated. Here, we report that insulin/insulin-like growth factor signaling in adipocytes promotes GSC maintenance, early germline cyst survival, and vitellogenesis. Further, adipocytes use distinct mechanisms downstream of insulin receptor activation to control these aspects of oogenesis, all of which are independent of FOXO. We find that GSC maintenance is modulated by Akt1 through GSK-3ß, early germline cyst survival is downstream of adipocyte Akt1 but independent of GSK-3ß, and vitellogenesis is regulated through an Akt1-independent pathway in adipocytes. These results indicate that, in addition to employing different types of nutrient sensing, adipocytes can use distinct axes of a single nutrient-sensing pathway to regulate multiple stages of the GSC lineage in the ovary.


Assuntos
Adipócitos/fisiologia , Células-Tronco Germinativas Adultas/fisiologia , Glicogênio Sintase Quinase 3 beta/metabolismo , Adipócitos/metabolismo , Células-Tronco Germinativas Adultas/metabolismo , Animais , Contagem de Células , Proliferação de Células , Drosophila/metabolismo , Proteínas de Drosophila/metabolismo , Proteínas de Drosophila/fisiologia , Feminino , Fatores de Transcrição Forkhead/metabolismo , Fatores de Transcrição Forkhead/fisiologia , Células Germinativas/citologia , Glicogênio Sintase Quinase 3 beta/fisiologia , Insulina/metabolismo , Masculino , Oogênese/fisiologia , Ovário/citologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/fisiologia , Células-Tronco/citologia
13.
J Assist Reprod Genet ; 35(7): 1265-1276, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29774457

RESUMO

PURPOSE: This study investigated the relationship between the vitamin D [25(OH)D] level in individual follicles and oocyte developmental competence. METHODS: A prospective cohort study in a private infertility center. Infertile women (N = 198) scheduled for intracytoplasmic sperm injection (ICSI) and a single embryo transfer (SET) provided serum samples and 322 follicular fluid (FF) specimens, each from a single follicle on the day of oocyte retrieval. RESULTS: FFs corresponding to successfully fertilized oocytes (following ICSI) contained significantly lower 25(OH)D level compared with those that were not fertilized (28.4 vs. 34.0 ng/ml, P = 0.001). Top quality embryos on the third day after fertilization, when compared to other available embryos, developed from oocytes collected from follicles containing significantly lower 25(OH)D levels (24.56 vs. 29.59 ng/ml, P = 0.007). Positive hCG, clinical pregnancy, and live birth rates were achieved from embryos derived from oocytes that grew in FF with significantly lower 25(OH)D levels than in follicles not associated with subsequent pregnancy. The concentration of 25(OH)D in FF in women with negative hCG was 32.23 ± 20.21 ng/ml, positive hCG 23.62 ± 6.09 ng/ml, clinical pregnancy 23.13 ± 6.09 ng/ml, and live birth 23.45 ± 6.11 ng/ml (P < 0.001). Women with serum 25(OH)D < 20 ng/ml had not only a higher fertilization rate (71 vs. 61.6%, P = 0.026) and a higher clinical pregnancy rate (48.2 vs. 25%, P = 0.001), but also higher miscarriage rate (14.5 vs. 3.8%, P = 0.013) compared with those with levels ≥ 20 ng/ml. CONCLUSION: This study reveals that the level of 25(OH)D in FF correlates negatively with the oocytes' ability to undergo fertilization and subsequent preimplantation embryo development. Oocytes matured in FF with low 25(OH)D concentration are more likely to produce top quality embryos and are associated with higher pregnancy and delivery rates. On the other hand, low serum vitamin D concentration is associated with higher miscarriage rates.


Assuntos
Biomarcadores/sangue , Oócitos/metabolismo , Oócitos/fisiologia , Folículo Ovariano/metabolismo , Vitamina D/sangue , Vitamina D/metabolismo , Adulto , Coeficiente de Natalidade , Desenvolvimento Embrionário/fisiologia , Feminino , Fertilização/fisiologia , Fertilização In Vitro/métodos , Líquido Folicular/metabolismo , Líquido Folicular/fisiologia , Humanos , Infertilidade Feminina/sangue , Infertilidade Feminina/metabolismo , Infertilidade Feminina/fisiopatologia , Nascimento Vivo , Recuperação de Oócitos/métodos , Oogênese/fisiologia , Folículo Ovariano/fisiologia , Gravidez , Taxa de Gravidez , Estudos Prospectivos , Injeções de Esperma Intracitoplásmicas/métodos
14.
Dev Biol ; 440(2): 99-112, 2018 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-29753016

RESUMO

Intercellular bridges are conserved structures that allow neighboring cells to exchange cytoplasmic material; defects in intercellular bridges can lead to infertility in many organisms. Here, we use the Drosophila egg chamber to study the mechanisms that regulate intercellular bridges. Within the developing egg chamber, the germ cells (15 nurse cells and 1 oocyte) are connected to each other through intercellular bridges called ring canals, which expand over the course of oogenesis to support the transfer of materials from the nurse cells to the oocyte. The ring canals are enriched in actin and actin binding proteins, and many proteins have been identified that localize to the germline ring canals and control their expansion and stability. Here, we demonstrate a novel role for the Ste20 family kinase, Misshapen (Msn), in regulation of the size of the germline ring canals. Msn localizes to ring canals throughout most of oogenesis, and depletion of Msn led to the formation of larger ring canals. Over-expression of Msn decreased ring canal diameter, and expression of a membrane tethered form of Msn caused ring canal detachment and nurse cell fusion. Altering the levels or localization of Msn also led to changes in the actin cytoskeleton and altered the localization of E-cadherin, which suggests that Msn could be indirectly limiting ring canal size by altering the structure or dynamics of the actin cytoskeleton and/or adherens junctions.


Assuntos
Proteínas de Drosophila/metabolismo , Proteínas de Drosophila/fisiologia , Células Germinativas/fisiologia , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Serina-Treonina Quinases/fisiologia , Citoesqueleto de Actina/metabolismo , Actinas/metabolismo , Animais , Proteínas Contráteis/metabolismo , Drosophila/genética , Drosophila/metabolismo , Proteínas de Drosophila/genética , Feminino , Células Germinativas/citologia , Células Germinativas/metabolismo , Proteínas dos Microfilamentos/metabolismo , Oócitos/citologia , Oócitos/enzimologia , Oogênese/fisiologia , Proteínas Serina-Treonina Quinases/genética
15.
J Assist Reprod Genet ; 35(7): 1135-1148, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29691711

RESUMO

PURPOSE: Mammalian oogenesis and folliculogenesis share a dynamic connection that is critical for gamete development. For maintenance of quiescence or follicular activation, follicles must respond to soluble signals (growth factors and hormones) and physical stresses, including mechanical forces and osmotic shifts. Likewise, mechanical processes are involved in cortical tension and cell polarity in oocytes. Our objective was to examine the contribution and influence of biomechanical signaling in female mammalian gametogenesis. METHODS: We performed a systematic review to assess and summarize the effects of mechanical signaling and mechanotransduction in oocyte maturation and folliculogenesis and to explore possible clinical applications. The review identified 2568 publications of which 122 met the inclusion criteria. RESULTS: The integration of mechanical and cell signaling pathways in gametogenesis is complex. Follicular activation or quiescence are influenced by mechanical signaling through the Hippo and Akt pathways involving the yes-associated protein (YAP), transcriptional coactivator with PDZ-binding motif (TAZ), phosphatase and tensin homolog deleted from chromosome 10 (PTEN) gene, the mammalian target of rapamycin (mTOR), and forkhead box O3 (FOXO3) gene. CONCLUSIONS: There is overwhelming evidence that mechanical signaling plays a crucial role in development of the ovary, follicle, and oocyte throughout gametogenesis. Emerging data suggest the complexities of mechanotransduction and the biomechanics of oocytes and follicles are integral to understanding of primary ovarian insufficiency, ovarian aging, polycystic ovary syndrome, and applications of fertility preservation.


Assuntos
Mecanotransdução Celular/fisiologia , Oogênese/fisiologia , Ovário/fisiologia , Transdução de Sinais/fisiologia , Animais , Feminino , Humanos , Oócitos/fisiologia , Folículo Ovariano/fisiologia
16.
Zygote ; 26(2): 168-176, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29607795

RESUMO

SummaryThis study aimed to understand how germ cell development occurs in females of Devario aequipinnatus, by morphologically describing oogenesis and the reproductive phases. Sexually mature females of D. aequipinnatus (n = 70) were obtained from commercial fisheries and delivered to the Laboratório de Ictiologia Neotropical, UNESP, Ilha Solteira, SP, Brazil. The ovaries were removed, fragmented and fixed following the usual techniques for light microscopy. The stages of ovarian development in D. aequipinnatus begin with the oogonia, which proliferate into new cells or differentiate into prophasic oocytes that, at the end of this process, form the ovarian follicle and end folliculogenesis. In the previtellogenic stage, the oocytes were characterized mainly by the gradual loss of basophilia and an increase in oocyte diameter. Vitellogenesis was marked mainly by the incorporation of yolk granules. Mature oocytes were defined by their migration from the nucleus to the micropyle. Postovulatory follicles and atresic oocytes were also observed. The reproductive phases were classified as: immature, early and final developing, spawning capable, regressing and regenerating. Therefore, the development of an understanding of cell modifications that occurs up to oogenesis is a basic step that is essential for the description of the reproductive biology of D. aequipinnatus, given the lack of information about the reproductive aspects of this species.


Assuntos
Cyprinidae/fisiologia , Ciclo Menstrual/fisiologia , Oócitos/fisiologia , Animais , Feminino , Oócitos/citologia , Oogênese/fisiologia , Ovário/citologia , Ovário/fisiologia
17.
C R Biol ; 341(4): 219-227, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29705198

RESUMO

At the beginning of diplotene, the oocyte of Xenopus laevis is a cell of about 10-20 microns destined to increase 10,000-fold its size when the oocyte becomes filled with yolk platelets and has accumulated a great number of pigment granules in a half of its periphery. Its internal architecture is gradually accomplished during growth because of several factors, especially because of cytoskeletal changes. In the fully-grown oocyte, the cytoskeleton appears to sustain the eccentrically located germinal vesicle through arms radiating from the cortex to the germinal vesicle, a unique organization not to be found in other Amphibians. In this report, we summarized and analysed steps of cytoskeletal proteins and related mRNAs organization and function throughout diplotene stage, highlighting our studies in this animal model. The cytoskeletal proteins appear to exploit their activity with respect to ribosomal 60S subunit maturation and during translation. Most importantly, the polarity of the oocyte is achieved through a sophisticated and highly organized localization of mRNAs and cytoskeletal proteins in one side of the cell. This asymmetry will start the construction of the oocyte polarity that is instrumental for determining the characteristic of this cell, which will become an embryo. Moreover, in the same time membrane composition, conditioned by the underlying cytoskeletal organization, will acquire the prerequisites for sperm binding and fusion.


Assuntos
Citoesqueleto/fisiologia , Oócitos/metabolismo , Xenopus laevis , Animais , Citoplasma/fisiologia , Feminino , Microtúbulos/metabolismo , Oogênese/fisiologia , RNA Mensageiro
18.
Reprod Biomed Online ; 37(1): 25-32, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29703434

RESUMO

RESEARCH QUESTION: Are miRNAs found in follicular fluid related to blastocyst formation from the corresponding oocytes? DESIGN: In this study, 91 individual follicular fluid samples from single follicles containing mature oocytes from 91 women were collected and classified into group 1 (n = 38) with viable blastocysts, and group 2 (n = 53) with no blastocyst. TaqMan human miRNA cards and quantitative reverse transcription polymerase chain reaction were used to identify differently expressed follicular fluid miRNAs between the two groups. RESULTS: We found MIR-663B to be significantly differentially expressed in follicular fluid of oocytes that yielded viable blastocysts versus those that did not develop into blastocysts (14.16 ± 7.00 versus 23.68 ± 17.02; P = 0.019), as well as for those which develop into blastocysts with good morphology versus those with poor morphology (11.69 ± 3.49 versus 20.16 ± 9.33; P = 0.003). CONCLUSIONS: MIR-663B expression levels in human follicular fluid samples were significantly negatively related to viable blastocyst formation and may become an objective evaluation criterion for embryo development potential after IVF.


Assuntos
Blastocisto/metabolismo , Desenvolvimento Embrionário/genética , Líquido Folicular/metabolismo , MicroRNAs/metabolismo , Adulto , Feminino , Humanos , MicroRNAs/genética , Oócitos/metabolismo , Oogênese/fisiologia , Injeções de Esperma Intracitoplásmicas
19.
PLoS One ; 13(4): e0195647, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29694411

RESUMO

Matrotrophic viviparity is a reproductive pattern in which offspring develop inside a female's body which provides gas exchange and nutrients necessary for development. Besides placental mammals, structural and physiological aspects of matrotrophic viviparity are poorly characterized. In insects, the majority of species is oviparous, i.e. lay eggs, and viviparous reproduction has been reported only in 11 out of 44 orders, including earwigs (Dermaptera). Among dermapterans, matrotrophic viviparity has been reported in two epizoic subgroups: Arixeniidae and Hemimeridae. Here, we provide morphological evidence for distinct adaptations for this mode of viviparity in embryonic and maternal tissues in a representative of the latter subgroup, Hemimerus talpoides. Our study reveals a novel mechanism of maternal contribution to embryonic development which operates during oogenesis and involves characteristic modification of endoplasmic reticulum cisternae. Conspicuous and apparently inactive para-crystalline stacks of the endoplasmic reticulum are deposited in the oocyte cytoplasm and become activated during early embryonic development. Our analyses indicate additionally that in Hemimerus, transformed follicular/ovarian cells (on the mother's side) and an evagination of the dorsal vessel (on the embryo's side) converge to form a cephalic vesicle, structure analogous to a placenta. The cellular architecture of this unusual "cephalic placenta" points to its participation in an exchange of low molecular weight substances between a mother and developing embryo.


Assuntos
Insetos/anatomia & histologia , Insetos/embriologia , Oócitos/ultraestrutura , Viviparidade não Mamífera , Adaptação Fisiológica , Animais , Retículo Endoplasmático/metabolismo , Insetos/fisiologia , Insetos/ultraestrutura , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Oócitos/metabolismo , Oogênese/fisiologia , Oviparidade/fisiologia , Ratos , Viviparidade não Mamífera/fisiologia
20.
Dev Biol ; 438(1): 1-9, 2018 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-29571611

RESUMO

Apoptosis not only eliminates cells that are damaged or dangerous but also cells whose function during development in patterning or organogenesis is complete. The successful formation of germ cells is essential for the perpetuation of a species. The production of an oocyte often depends on signaling between germline and somatic cells, but also between specialized types of somatic cells. In Drosophila, each developing egg chamber is separated from the next by a single file of interfollicular somatic cells. Little is known about the function of the interfollicular stalk, although its presumed role in separating egg chambers is to ensure that patterning cues from one egg chamber do not impact or disrupt the development of adjacent egg chambers. We found that cells comprising the stalk undergo a progressive decrease in number during oogenesis through an apoptotic-dependent loss. The extent of programmed cell death is restricted by JAK/STAT signaling in a cell-autonomous manner to ensure that the stalk is maintained. Both a failure to undergo the normal reduction in stalk cell number, or to prevent excessive stalk cell apoptosis results in a decrease in fecundity. Thus, activation of JAK/STAT signaling in the Drosophila interfollicular stalk emerges as a model to study the tight regulation of signaling-dependent apoptosis.


Assuntos
Apoptose/genética , Janus Quinases/metabolismo , Oogênese/genética , Ovário/citologia , Fatores de Transcrição STAT/metabolismo , Animais , Contagem de Células , Drosophila/metabolismo , Proteínas de Drosophila/metabolismo , Feminino , Imuno-Histoquímica , Oogênese/fisiologia , Folículo Ovariano/citologia , Folículo Ovariano/fisiologia , Ovário/metabolismo , Ovário/fisiologia , Transdução de Sinais/fisiologia
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