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1.
Genes Dev ; 34(3-4): 166-178, 2020 02 01.
Artículo en Inglés | MEDLINE | ID: mdl-31919188

RESUMEN

Oocytes are indispensable for mammalian life. Thus, it is important to understand how mature oocytes are generated. As a critical stage of oocytes development, meiosis has been extensively studied, yet how chromatin remodeling contributes to this process is largely unknown. Here, we demonstrate that the ATP-dependent chromatin remodeling factor Snf2h (also known as Smarca5) plays a critical role in regulating meiotic cell cycle progression. Females with oocyte-specific depletion of Snf2h are infertile and oocytes lacking Snf2h fail to undergo meiotic resumption. Mechanistically, depletion of Snf2h results in dysregulation of meiosis-related genes, which causes failure of maturation-promoting factor (MPF) activation. ATAC-seq analysis in oocytes revealed that Snf2h regulates transcription of key meiotic genes, such as Prkar2b, by increasing its promoter chromatin accessibility. Thus, our studies not only demonstrate the importance of Snf2h in oocyte meiotic resumption, but also reveal the mechanism underlying how a chromatin remodeling factor can regulate oocyte meiosis.


Asunto(s)
Adenosina Trifosfatasas/genética , Adenosina Trifosfatasas/metabolismo , Ensamble y Desensamble de Cromatina/genética , Proteínas Cromosómicas no Histona/genética , Proteínas Cromosómicas no Histona/metabolismo , Factor Promotor de Maduración/genética , Meiosis/genética , Oogénesis/genética , Animales , Femenino , Regulación del Desarrollo de la Expresión Génica , Técnicas de Inactivación de Genes , Ratones , Oocitos/citología , Transcriptoma
2.
Results Probl Cell Differ ; 68: 477-494, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31598868

RESUMEN

The subphylum Chelicerata represents one of the oldest groups among arthropods and comprises more than a dozen orders. Representatives of particular orders differ significantly in their external morphology, reproductive biology, behavior, and structure of internal organs, e.g. of the respiratory system. However, in almost all chelicerates (excluding some mites) the female gonads show a similar architecture. In this chapter, the chelicerate-type ovary structure and the course of oogenesis are described. Structural and functional diversities of the chelicerate-type ovary in non-matrotrophic and matrotrophic arachnids are also presented.


Asunto(s)
Artrópodos/anatomía & histología , Artrópodos/citología , Oogénesis , Ovario/anatomía & histología , Ovario/citología , Animales , Arácnidos/anatomía & histología , Arácnidos/citología , Femenino
3.
Results Probl Cell Differ ; 68: 495-513, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31598869

RESUMEN

Even though tardigrades have been known since 1772, their phylogenetic position is still controversial. Tardigrades are regarded as either the sister group of arthropods, onychophorans, or onychophorans plus arthropods. Furthermore, the knowledge about their gametogenesis, especially oogenesis, is still poor and needs further analysis. The process of oogenesis has been studied solely for several eutardigradan species. Moreover, the spatial organization of the female germ-line clusters has been described for three species only. Meroistic ovaries characterize all analyzed species. In species of the Parachela, one cell per germ-cell cluster differentiates into the oocyte, while the remaining cells become the trophocytes. In Apochela several cells in the cluster differentiate into oocytes. Vitellogenesis is of a mixed type. The eggs are covered with the egg capsule that is composed of two shells: the thin vitelline envelope that adheres to the oolemma and the thick three-layered chorion. Chorion is formed as a first followed by vitelline envelope. Several features related to the oogenesis and structure of the ovary confirm the hypothesis that tardigrades are the sister group rather for arthropods than for onychophorans.


Asunto(s)
Oocitos/citología , Oogénesis , Ovario/anatomía & histología , Tardigrada/anatomía & histología , Tardigrada/fisiología , Animales , Femenino , Ovario/citología , Filogenia , Tardigrada/clasificación
4.
Results Probl Cell Differ ; 68: 515-551, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31598870

RESUMEN

Animal female and male germ-line cells often form syncytial units termed cysts, clusters, or clones. Within these cysts, the cells remain interconnected by specific cell junctions known as intercellular bridges or ring canals, which enable cytoplasm to be shared and macromolecules and organelles to be exchanged between cells. Numerous analyses have shown that the spatial organization of cysts and their functioning may differ between the sexes and taxa. The vast majority of our knowledge about the formation and functioning of germ-line cysts comes from studies of model species (mainly Drosophila melanogaster); the other systems of the cyst organization and functioning are much less known and are sometimes overlooked. Here, we present the current state of the knowledge of female germ-line cysts in clitellate annelids (Clitellata), which is a monophyletic taxon of segmented worms (Annelida). The organization of germ-line cysts in clitellates differs markedly from that of the fruit fly and vertebrates. In Clitellata, germ cells are not directly connected one to another, but, as a rule, each cell has one ring canal that connects it to an anuclear central cytoplasmic core, a cytophore. Thus, this pattern of cell distribution is similar to the germ-line cysts of Caenorhabditis elegans. The last decade of studies has revealed that although clitellate female germ-line cysts have a strong morphological plasticity, e.g., cysts may contain from 16 to as many as 2500 cells, the oogenesis always shows a meroistic mode, i.e., the interconnected cells take on different fates; a few (sometimes only one) become oocytes, whereas the rest play the role of supporting (nurse) cells and do not continue oogenesis.This is the first comprehensive summary of the current knowledge on the organization and functioning of female germ-line cysts in clitellate annelids.


Asunto(s)
Anélidos/citología , Células Germinativas/citología , Células Gigantes/citología , Células Gigantes/fisiología , Animales , Femenino , Oocitos/citología , Oocitos/crecimiento & desarrollo , Oogénesis
5.
Environ Pollut ; 255(Pt 1): 113194, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31520902

RESUMEN

Heavy metal cadmium (Cd) is a widespread environmental contaminant with a potential toxicity that might adversely influence the health of experimental animals and humans. It has been known that Cd might accumulate in vertebrates for many years and thus leads to the hepatic and renal toxicity. Additionally, Cd concentration in the ovary increases with age and is highly related to the reproductive hazard. However, the underlying mechanisms regarding how Cd affects the female reproductive system especially the oocyte quality have not yet fully defined. Here, we reported that Cd exposure led to the defective nuclear maturation of oocytes via the impairment of cytoskeleton assembly, displaying the aberrant spindle organization, chromosome alignment and actin polymerization. In the meantime, Cd exposure caused the impaired cytoplasmic maturation by showing the disrupted dynamics of mitochondrial integrity and cortical granules, and thereby resulting in the compromised sperm binding ability and fertilization capacity of oocytes. More importantly, we found that glutathione (GSH) supplementation was able to recover the meiotic failure induced by Cd exposure through suppressing the excessive ROS level, DNA damage accumulation and apoptotic incidence. Taken together, our findings demonstrate that Cd exposure has the adverse effects on the oocyte meiotic maturation as well as subsequent fertilization, and provide a potential effective strategy to improve the quality of Cd-exposed oocytes.


Asunto(s)
Cadmio/toxicidad , Mitocondrias/patología , Oocitos/citología , Oogénesis/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Animales , Citoesqueleto/efectos de los fármacos , Daño del ADN/efectos de los fármacos , Femenino , Glutatión/metabolismo , Humanos , Masculino , Meiosis/efectos de los fármacos , Oocitos/patología , Porcinos
6.
Adv Exp Med Biol ; 1169: 213-223, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31487026

RESUMEN

Every organ in the body is thought to harbor two populations of stem cells, including the quiescent and the actively dividing, that leads to heterogeneity among them. It is generally believed that the ovary harbors a fixed number of follicles at birth that differentiate during fetal development from the primordial germ cells. The numbers of follicles decrease by age, leading to menopause. However, in 2004, it was suggested that ovary may harbor stem cells that are possibly involved in the formation of new follicles throughout reproductive life. Research over little more than a decade shows that ovarian stem cells include a quiescent population of very small embryonic-like stem cells (VSELs) and slightly bigger, actively dividing ovarian stem cells (OSCs). This heterogeneity among ovarian stem cells is similar to the presence of VSELs along with spermatogonial stem cells (SSCs) in the testis or hematopoietic stem cells (HSCs) in the hematopoietic system. VSELs express embryonic markers, including nuclear OCT-4, and are lodged in the ovary surface epithelium (OSE). Ovarian VSELs undergo asymmetric cell division to self-renew and give rise to OSCs that in turn undergo symmetric cell divisions and clonal expansion (germ cell nest) followed by meiosis to form an oocyte that gets assembled as a primordial follicle. Both VSELs and OSCs also express receptors for follicle-stimulating hormone (FSHR) and are directly activated by FSH to undergo neo-oogenesis and primordial follicle assembly. Whether stimulation of ovaries by FSH in Infertility Clinics activates the stem cells leading to the formation of multiple follicles needs further investigation. Epithelial cells lining the surface of ovary provide a niche to the stem cells under normal circumstances and undergo epithelial-mesenchymal transition (EMT) to form granulosa cells for primordial follicle assembly. Compromised function of the epithelial cells with age possibly leads to inability of stem cells to form follicles, leading to menopause. More than 90% of ovarian cancers arise in the OSE, possibly due to excessive self-renewal of VSELs. Altered biology of the OSE cells results in the formation of myofibroblasts by EMT and may provide a cancerous niche that supports excessive expansion of the stem cells lodged in the OSE, leading to ovarian cancer. Ovarian cancer cells express markers like OCT-4 and FSHR, which are also expressed by the VSELs lodged in the OSE, whereas the epithelial cells are distinctly negative for the same. Lot more research is required in the field to gain further understanding of ovarian stem cell biology.


Asunto(s)
Células Madre Embrionarias , Folículo Ovárico , Células Madre Embrionarias/citología , Femenino , Humanos , Masculino , Oogénesis , Folículo Ovárico/citología
7.
Nat Commun ; 10(1): 3940, 2019 09 02.
Artículo en Inglés | MEDLINE | ID: mdl-31477736

RESUMEN

Collective cell migration is involved in development, wound healing and metastasis. In the Drosophila ovary, border cells (BC) form a small cluster that migrates collectively through the egg chamber. To achieve directed motility, the BC cluster coordinates the formation of protrusions in its leader cell and contractility at the rear. Restricting protrusions to leader cells requires the actin and plasma membrane linker Moesin. Herein, we show that the Ste20-like kinase Misshapen phosphorylates Moesin in vitro and in BC. Depletion of Misshapen disrupts protrusion restriction, thereby allowing other cells within the cluster to protrude. In addition, we show that Misshapen is critical to generate contractile forces both at the rear of the cluster and at the base of protrusions. Together, our results indicate that Misshapen is a key regulator of BC migration as it coordinates two independent pathways that restrict protrusion formation to the leader cells and induces contractile forces.


Asunto(s)
Actomiosina/genética , Movimiento Celular/genética , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Regulación del Desarrollo de la Expresión Génica , Oogénesis/genética , Proteínas Serina-Treonina Quinasas/genética , Citoesqueleto de Actina/metabolismo , Actomiosina/metabolismo , Algoritmos , Animales , Animales Modificados Genéticamente , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Femenino , Proteínas de Microfilamentos/genética , Proteínas de Microfilamentos/metabolismo , Modelos Genéticos , Proteínas Serina-Treonina Quinasas/metabolismo , Interferencia de ARN
8.
Zygote ; 27(5): 329-336, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31412969

RESUMEN

Mammalian Pou5f1 encodes the POU family class V (POU-V) transcription factor which is essential for the pluripotency of embryonic cells and germ cells. In vertebrates, various POU-V family genes have been identified and classified into the POU5F1 family or its paralogous POU5F3 family. In this study, we cloned two cDNAs named CpPou5f1 and CpPou5f3, which encode POU-V family proteins of the Japanese red bellied newt Cynops pyrrhogaster. In the predicted amino acid sequence encoded by CpPou5f1, the typical MAGH sequence at the N-terminus and deletion of arginine at the fifth position of POU-homeodomain were recognized, but not in the sequence encoded by CpPou5f3. Phylogenetic analysis using Clustal Omega software indicated that CpPou5f1 and CpPou5f3 are classified into the clade of the POU5F1 and POU5F3 families, respectively. In a real-time polymerase chain reaction (RT-PCR) analysis, the marked gene expression of CpPou5f1 was observed during oogenesis and early development up to the tail-bud stage, whereas weak gene expression of CpPou5f3 was detected only in the early stages of oogenesis and gastrula. In adult organs, CpPou5f1 was expressed only in the ovary, while gene expression of CpPou5f3 was recognized in various organs. A regeneration experiment using larval forelimb revealed that transient gene expression of CpPou5f1 occurred at the time of wound healing, followed by gene activation of CpPou5f3 during the period of blastema formation. These results suggest that CpPou5f1 and CpPou5f3 might play different roles in embryogenesis and limb regeneration.


Asunto(s)
Oogénesis/genética , Factores del Dominio POU/genética , Regeneración/genética , Salamandridae/genética , Animales , Embrión no Mamífero/fisiología , Extremidades/fisiología , Femenino , Regulación del Desarrollo de la Expresión Génica , Masculino , Familia de Multigenes , Filogenia , Salamandridae/embriología , Salamandridae/fisiología
9.
Nat Commun ; 10(1): 3858, 2019 08 26.
Artículo en Inglés | MEDLINE | ID: mdl-31451685

RESUMEN

The Polycomb group of proteins is required for the proper orchestration of gene expression due to its role in maintaining transcriptional silencing. It is composed of several chromatin modifying complexes, including Polycomb Repressive Complex 2 (PRC2), which deposits H3K27me2/3. Here, we report the identification of a cofactor of PRC2, EZHIP (EZH1/2 Inhibitory Protein), expressed predominantly in the gonads. EZHIP limits the enzymatic activity of PRC2 and lessens the interaction between the core complex and its accessory subunits, but does not interfere with PRC2 recruitment to chromatin. Deletion of Ezhip in mice leads to a global increase in H3K27me2/3 deposition both during spermatogenesis and at late stages of oocyte maturation. This does not affect the initial number of follicles but is associated with a reduction of follicles in aging. Our results suggest that mature oocytes Ezhip-/- might not be fully functional and indicate that fertility is strongly impaired in Ezhip-/- females. Altogether, our study uncovers EZHIP as a regulator of chromatin landscape in gametes.


Asunto(s)
Proteínas Oncogénicas/metabolismo , Óvulo/metabolismo , Complejo Represivo Polycomb 2/metabolismo , Espermatozoides/metabolismo , Adulto , Animales , Línea Celular Tumoral , Cromatina/metabolismo , Femenino , Células HEK293 , Histonas/metabolismo , Humanos , Masculino , Ratones , Ratones Noqueados , Mutación , Proteínas Oncogénicas/genética , Proteínas Oncogénicas/aislamiento & purificación , Oogénesis , Ovario/citología , Ovario/patología , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo , Células Sf9 , Espermatogénesis , Testículo/citología , Testículo/patología
10.
Chemosphere ; 237: 124435, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31352102

RESUMEN

Glyphosate is a high-efficiency, low-toxicity, broad-spectrum herbicide. The residues of glyphosate-based herbicides are frequent pollutants in the environment. However, the effects of glyphosate on oocyte maturation, as well as its possible mechanisms, remain unclear. The present study revealed that mouse oocytes had reduced rates of germinal vesicle breakdown (GVBD) and first polar body extrusion (PBE) after treatment with 500 µM glyphosate. Reactive oxygen species (ROS) were found in mouse oocytes exposed to glyphosate, as shown by changes in the mRNA expression of related antioxidant enzyme genes (cat, sod2, gpx). After 14 h of exposure to glyphosate, metaphase II (MII) mouse oocytes displayed an abnormal spindle morphology and DNA double-strand breaks (DNA-DSBs). Simultaneously, mitochondria showed an aggregated distribution and decreased membrane potential in mouse oocytes exposed to glyphosate. The protein expression levels of apoptosis factors (Bax, Bcl-2) and the mRNA expression levels of apoptosis-related genes (bax, bcl-2, caspase3) were measured by Western blot and qRT-PCR, respectively. Meanwhile, the expression levels of autophagy-related genes (lc3, atg14, mtor) and proteins (LC3, Atg12) were significantly decreased in the glyphosate treatment group compared with the control group. Collectively, our results indicated that glyphosate exposure could interfere with mouse oocyte maturation by generating oxidative stress and early apoptosis.


Asunto(s)
Glicina/análogos & derivados , Oocitos/efectos de los fármacos , Animales , Apoptosis/efectos de los fármacos , Autofagia/efectos de los fármacos , Femenino , Glicina/toxicidad , Herbicidas/toxicidad , Ratones , Mitocondrias/metabolismo , Oogénesis/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo
11.
Gen Comp Endocrinol ; 282: 113218, 2019 10 01.
Artículo en Inglés | MEDLINE | ID: mdl-31301284

RESUMEN

Progestin receptor membrane component (Pgrmc1 & 2) is a heme-binding protein. Studies on Pgrmc1 have suggested possible roles in heme binding, activation of steroid-synthesizing P450s, along with binding and transferring of membrane proteins. However, the studies of Pgrmc1's paralog, Pgrmc2 are still lacking. In order to determine the physiologic function(s) of Pgrmc2, we generated a zebrafish mutant line (pgrmc2-/-). We found a reduction in both spawning frequency and the number of embryos produced in female pgrmc2-/-. This subfertility is caused by reduced oocyte maturation (germinal vesicle breakdown, GVBD) in pgrmc2-/- in vivo. Nonetheless, oocytes from pgrmc2-/- had similar sensitivity to 17α,20ß-dihydroxy-4-pregnen-3-one (DHP, a maturation induced progestin in zebrafish) compared with wildtype (wt) in vitro. Therefore, we hypothesized that oocyte maturation tardiness found in vivo, could be due to lack of progestin in pgrmc2-/-. Interestingly, we found significant reduced expression of hormones, receptors, and steroid synthesizing enzymes including lhcgr, egfra, ar, and esr2, cyp11a1 and hsd3b1. In addition, DHP levels in pgrmc2-/- ovaries showed a significant decrease compared to those in wt. In summary, we have provided a plausible molecular mechanism for the physiological functions of Pgrmc2 in the regulation of female fertility, likely via regulation of receptors and steroids in the ovary, which in turn regulates oocyte maturation in zebrafish.


Asunto(s)
Infertilidad/metabolismo , Infertilidad/patología , Progestinas/biosíntesis , Proteínas de Pez Cebra/metabolismo , Pez Cebra/fisiología , Animales , Secuencia de Bases , Femenino , Regulación de la Expresión Génica , Mutación/genética , Oocitos/metabolismo , Oogénesis , Folículo Ovárico/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reproducción/genética , Maduración Sexual , Pez Cebra/metabolismo , Proteínas de Pez Cebra/deficiencia , Proteínas de Pez Cebra/genética
12.
In Vitro Cell Dev Biol Anim ; 55(7): 548-558, 2019 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-31313007

RESUMEN

Recently, the mean maternal age at first birth has been continuing to increase. The decline in the age-related fertility is due to the reduction in the number and the quality of the oocyte. An elevation in intra-ovarian reactive oxygen species (ROS) is correlated with the increase in maternal age, and the oxidative stress is involved in the decline in oocyte quality. Although ß-carotene, a very effective quencher of ROS, has been found to have the beneficial contribution to the ovarian development and steroidogenesis, it is unknown the effect of ß-carotene on the oocyte development especially oocyte maturation. This investigation aimed to explore the beneficial contribution of ß-carotene on oocyte maturation under oxidative stress and the underlying mechanism. We found that the oxidative stress induced by ROS reagent Rosup inhibited oocyte development/maturation and parthenogenetic activation which could be dramatically rescued by ß-carotene (57.1 ± 4.7% vs 78.9 ± 3.8%; p < 0.05) in vitro. The underlying mechanisms include that ß-carotene not only reduces ROS formation and cell apoptosis, but also it can restore actin expression, cortical granule-free domain (CGFD) formation, mitochondria homogeneous distribution, and nuclear maturation. The data suggest that ß-carotene acts as a potential antioxidant in the oocyte. Therefore, the findings from this investigation provide the fundamental 7knowledge for using ß-carotene as an antioxidant to improve the oocyte quality and even the ovarian function.


Asunto(s)
Antioxidantes/farmacología , Oocitos/citología , Oogénesis/fisiología , Estrés Oxidativo/efectos de los fármacos , beta Caroteno/farmacología , Actinas/biosíntesis , Animales , Apoptosis/efectos de los fármacos , Células Cultivadas , Femenino , Edad Materna , Ratones , Mitocondrias/fisiología , Especies Reactivas de Oxígeno/metabolismo
13.
BMC Genomics ; 20(1): 588, 2019 Jul 17.
Artículo en Inglés | MEDLINE | ID: mdl-31315563

RESUMEN

BACKGROUND: Maturation of oocytes under in vitro conditions (IVM) results in impaired developmental competence compared to oocytes matured in vivo. As oocytes are closely coupled to their cumulus complex, elucidating aberrations in cumulus metabolism in vitro is important to bridge the gap towards more physiological maturation conditions. The aim of this study was to analyze the equine "cumulome" in a novel combination of proteomic (nano-HPLC MS/MS) and metabolomic (UPLC-nanoESI-MS) profiling of single cumulus complexes of metaphase II oocytes matured either in vivo (n = 8) or in vitro (n = 7). RESULTS: A total of 1811 quantifiable proteins and 906 metabolic compounds were identified. The proteome contained 216 differentially expressed proteins (p ≤ 0.05; FC ≥ 2; 95 decreased and 121 increased in vitro), and the metabolome contained 108 metabolites with significantly different abundance (p ≤ 0.05; FC ≥ 2; 24 decreased and 84 increased in vitro). The in vitro "cumulome" was summarized in the following 10 metabolic groups (containing 78 proteins and 21 metabolites): (1) oxygen supply, (2) glucose metabolism, (3) fatty acid metabolism, (4) oxidative phosphorylation, (5) amino acid metabolism, (6) purine and pyrimidine metabolism, (7) steroid metabolism, (8) extracellular matrix, (9) complement cascade and (10) coagulation cascade. The KEGG pathway "complement and coagulation cascades" (ID4610; n = 21) was significantly overrepresented after in vitro maturation. The findings indicate that the in vitro condition especially affects central metabolism and extracellular matrix composition. Important candidates for the metabolic group oxygen supply were underrepresented after maturation in vitro. Additionally, a shift towards glycolysis was detected in glucose metabolism. Therefore, under in vitro conditions, cumulus cells seem to preferentially consume excess available glucose to meet their energy requirements. Proteins involved in biosynthetic processes for fatty acids, cholesterol, amino acids, and purines exhibited higher abundances after maturation in vitro. CONCLUSION: This study revealed the marked impact of maturation conditions on the "cumulome" of individual cumulus oocyte complexes. Under the studied in vitro milieu, cumulus cells seem to compensate for a lack of important substrates by shifting to aerobic glycolysis. These findings will help to adapt culture media towards more physiological conditions for oocyte maturation.


Asunto(s)
Caballos/metabolismo , Técnicas de Maduración In Vitro de los Oocitos , Oocitos/metabolismo , Oogénesis , Animales , Células Cultivadas , Células del Cúmulo/metabolismo , Femenino , Metaboloma , Proteoma
14.
BMC Dev Biol ; 19(1): 14, 2019 07 05.
Artículo en Inglés | MEDLINE | ID: mdl-31277577

RESUMEN

BACKGROUND: Insulin/insulin-like growth peptide signaling (IIS) down-regulates hemolymph sugar level and facilitates larval growth in the soybean pod borer, Maruca vitrata. The objective of this study is to determine whether IIS of M. vitrata can mediate ovarian development of adult females. RESULTS: A pair of ovaries consists of 8 ovarioles, each of which is separated into distal germarium and proximal vitellarium in M. vitrata. In the germarium, oocyte development occurred with active mitotic activity which was visible by incorporating bromodeoxyribose uridine. Previtellogenic development and subsequent vitellogenesis began soon after adult emergence. They continued with increase of female age. Oocyte development was facilitated by up-regulation of vitellogenin (Vg) and Vg receptor (VgR) gene expression. Larval diets significantly influenced on ovarian development of M. vitrata because oocyte development varied with pupal size derived from larvae treated with different nutritional diets. Its ovarian development was dependent on endocrine signal(s) from the head because decapitation soon after adult emergence prevented oogenesis and subsequent vitellogenesis along with marked reduction of Vg and VgR expression. Topical application of juvenile hormone (JH) significantly recovered its ovarian development whereas farnesoic acid (a precursor of JH biosynthesis) or 20-hydroxyecdysone treatment did not. JH stimulated vitellogenesis and choriogenesis, but not previtellogenic development. In contrast, insulin injection to decapitated females stimulated oocyte differentiation and vitellogenesis along with increase of Vg and VgR expression. To further analyze the effect of insulin on ovarian development, expression of four IIS components (InR, FOXO, Akt, and TOR) genes was manipulated by RNA interference. Hemocoelic injection of gene-specific double stranded RNAs significantly reduced their target gene mRNA levels and interfered with ovarian development. An addition of insulin to JH treatment against decapitated females enhanced the gonadotropic effect of JH by stimulating oogenesis. CONCLUSIONS: IIS plays crucial role in mediating previtellogenic development of M. vitrata in response to nutrient signal. It also enhances the gonadotropic effect of JH II on vitellogenesis.


Asunto(s)
Factor I del Crecimiento Similar a la Insulina/metabolismo , Insulina/metabolismo , Hormonas Juveniles/metabolismo , Ovario/crecimiento & desarrollo , Vitelogénesis/fisiología , Animales , Ecdisterona/farmacología , Proteínas del Huevo/genética , Proteínas del Huevo/metabolismo , Ácidos Grasos Insaturados/farmacología , Femenino , Proteína Forkhead Box O1/genética , Mariposas Nocturnas , Oogénesis/fisiología , Proteínas Proto-Oncogénicas c-akt/genética , Interferencia de ARN , ARN Interferente Pequeño/genética , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/metabolismo , Receptores de Somatomedina/genética , Serina-Treonina Quinasas TOR/genética , Vitelogeninas/genética , Vitelogeninas/metabolismo
15.
J Assist Reprod Genet ; 36(7): 1457-1469, 2019 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-31187330

RESUMEN

PURPOSE: To determine whether a selected set of mRNA biomarkers expressed in individual cumulus granulosa cell (CC) masses show association with oocyte developmental competence, embryo ploidy status, and embryo outcomes. METHODS: This prospective observational cohort pilot study assessed levels of mRNA biomarkers in 163 individual CC samples from 15 women stimulated in antagonist cycles. Nineteen mRNA biomarker levels were measured by real-time PCR and related to the development of their corresponding individually cultured oocytes and subsequent embryos, embryo ploidy status, and live birth outcomes. RESULTS: PAPPA mRNA levels were significantly higher in CC from oocytes that led to euploid embryos resulting in live births and aneuploid embryos compared to immature oocytes by ANOVA. LHCGR mRNA levels were significantly higher in CC of oocytes resulting in embryos associated with live birth compared to immature oocytes and oocytes resulting in arrested embryos by ANOVA. Using a general linearized mixed model to assess ploidy status, CC HSD3B mRNA levels in oocytes producing euploid embryos were significantly lower than other oocyte outcomes, collectively. When transferred euploid embryos outcomes were analyzed by ANOVA, AREG mRNA levels were significantly lower and PAPPA mRNA levels significantly higher in CC from oocytes that produced live births compared to transferred embryos that did not form a pregnancy. CONCLUSIONS: Collectively, PAPPA, LHCGR, and AREG mRNA levels in CC may be able to identify oocytes with the best odds of resulting in a live birth, and HSD3B1 mRNA levels may be able to identify oocytes capable of producing euploid embryos.


Asunto(s)
Anfirregulina/genética , Complejos Multienzimáticos/genética , Oocitos/crecimiento & desarrollo , Proteína Plasmática A Asociada al Embarazo/genética , Progesterona Reductasa/genética , Receptores de HL/genética , Esteroide Isomerasas/genética , Adulto , Células del Cúmulo/metabolismo , Transferencia de Embrión , Desarrollo Embrionario/genética , Femenino , Regulación del Desarrollo de la Expresión Génica , Humanos , Técnicas de Maduración In Vitro de los Oocitos , Recuperación del Oocito , Oocitos/metabolismo , Oogénesis/genética , Folículo Ovárico/crecimiento & desarrollo , Folículo Ovárico/metabolismo , Ploidias , Embarazo , ARN Mensajero/genética
16.
Gen Comp Endocrinol ; 281: 83-90, 2019 09 15.
Artículo en Inglés | MEDLINE | ID: mdl-31170402

RESUMEN

The function of insulin-like growth factor (Igf) system in ovary has attracted much attention, but the role of Igf binding proteins (Igfbps) in ovary is still largely unknown. In this study, the role of Igfbps in oocyte maturation was investigated in zebrafish. The expression of all eight identified Igfbps except Igfbp6b could be detected in the adult ovary and exhibited differential expression profiles during folliculogenesis. The expression of several Igfbps is dynamically changed during oocyte maturation induced by human chorionic gonadotropin (hCG). By treatment of an Igfbps inhibitor NBI-31772 in vitro, the oocyte maturation could be stimulated in a clear dose-, time- and stage-dependent manner. Such effects were also observed by administration of NBI-31772 in vivo. Igfbps are differentially expressed in both follicular cells and oocytes, but the effect of NBI-31772 could only be found in intact follicles and not in the denuded oocytes. Previous studies have demonstrated that Igf3 is the major Igf member in regulating oocyte maturation of zebrafish. Interestingly, NBI-31772 could increase the effect of Igf3 on oocyte maturation. Furthermore, we found the effect of NBI-31772 on oocyte maturation could be blocked by an Igf type 1 receptor inhibitor BMS-536924 in vitro, suggesting the Igfbps can inhibit the oocyte maturation via Igf/Igf1r pathway. Together, we provided the first evidence in fish that Igfbps inhibit oocyte maturation of zebrafish.


Asunto(s)
Proteínas de Unión a Factor de Crecimiento Similar a la Insulina/metabolismo , Oocitos/fisiología , Proteínas de Pez Cebra/metabolismo , Pez Cebra/metabolismo , Animales , Catecoles/farmacología , Gonadotropina Coriónica/farmacología , Femenino , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Humanos , Proteínas de Unión a Factor de Crecimiento Similar a la Insulina/genética , Factor I del Crecimiento Similar a la Insulina/metabolismo , Isoquinolinas/farmacología , Oocitos/metabolismo , Oogénesis/efectos de los fármacos , Folículo Ovárico/citología , Folículo Ovárico/efectos de los fármacos , Folículo Ovárico/metabolismo , Receptor IGF Tipo 1/metabolismo , Transducción de Señal/efectos de los fármacos
17.
Zygote ; 27(3): 180-186, 2019 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-31171044

RESUMEN

SummaryHeat shock may disrupt oocyte function by increasing the generation of reactive oxygen species (ROS). We evaluated the capacity of the antioxidant melatonin to protect oocytes using two models of oxidative stress - heat shock and the pro-oxidant menadione. Bovine cumulus-oocyte complexes (COC) were exposed in the presence or absence of 1 µM melatonin to the following treatments during maturation: 38.5°C, 41°C and 38.5°C+5 µM menadione. In the first experiment, COC were matured for 3 h with 5 µM CellROX® and analyzed by epifluorescence microscopy to quantify production of ROS. The intensity of ROS was greater for oocytes exposed to heat shock and menadione than for control oocytes. Melatonin reduced ROS intensity for heat-shocked oocytes and oocytes exposed to menadione, but not for control oocytes. In the second experiment, COC were matured for 22 h. After maturation, oocytes were fertilized and the embryos cultured for 7.5 days. The proportion of oocytes that cleaved after fertilization was lower for oocytes exposed to heat shock and menadione than for control oocytes. Melatonin increased cleavage for heat-shocked oocytes and oocytes exposed to menadione, but not for control oocytes. Melatonin tended to increase the developmental competence of embryos from heat-shocked oocytes but not for embryos from oocytes exposed to menadione or from control oocytes. In conclusion, melatonin reduced production of ROS of maturing oocytes and protected oocytes from deleterious effects of both stresses on competence of the oocyte to cleave after coincubation with sperm. These results suggest that excessive production of ROS compromises oocyte function.


Asunto(s)
Respuesta al Choque Térmico , Melatonina/farmacología , Oocitos/efectos de los fármacos , Oogénesis/efectos de los fármacos , Estrés Oxidativo , Especies Reactivas de Oxígeno/metabolismo , Animales , Antioxidantes/farmacología , Bovinos , Femenino , Técnicas de Maduración In Vitro de los Oocitos , Microscopía Fluorescente , Oocitos/citología , Oocitos/metabolismo
18.
Fertil Steril ; 112(2): 387-396.e3, 2019 08.
Artículo en Inglés | MEDLINE | ID: mdl-31146888

RESUMEN

OBJECTIVE: To study whether increased body mass index is associated with altered expression of extracellular vesicle microRNAs (EV-linked miRNAs) in human follicular fluid. DESIGN: Cross-sectional study. SETTING: Tertiary-care university-affiliated center. PATIENT(S): One hundred thirty-three women undergoing in vitro fertilization (IVF) were recruited from January 2014 to August 2016. INTERVENTIONS(S): None. MAIN OUTCOME MEASURE(S): EV-linked miRNAs were isolated from follicular fluid and their expression profiles were measured with the use of the Taqman Open Array Human miRNA panel. EV-linked miRNAs were globally normalized and inverse-normal transformed. Associations between body mass index (BMI) and EV-linked miRNA outcomes were analyzed by means of multivariate linear regression and principal component analysis. RESULT(S): Eighteen EV-linked miRNAs were associated with an increase in BMI after adjusting for age, ethnicity, smoking status, and batch effects. Hsa-miR-328 remained significant after false discovery rate adjustments. Principal component analyses identified the first principal component to account for 40% of the variation in our EV-linked miRNA dataset, and adjusted linear regression found that the first principal component was significantly associated with BMI after multiple testing adjustments. Using Kyoto Encyclopedia of Genes and Genomes enrichment analyses, we predicted gene targets of EV-linked miRNA in silico and identified PI3K-Akt signaling, ECM-receptor interaction, focal adhesion, FoxO signaling, and oocyte meiosis pathways. CONCLUSION(S): These results show that a 1-unit increase in BMI is associated with altered follicular fluid expression of EV-linked miRNAs that may influence follicular and oocyte developmental pathways. Our findings provide potential insight into a mechanistic explanation for the reduced fertility rates associated with increased BMI.


Asunto(s)
Índice de Masa Corporal , Vesículas Extracelulares/metabolismo , Líquido Folicular/metabolismo , MicroARNs/metabolismo , Adulto , Estudios Transversales , Vesículas Extracelulares/genética , Femenino , Fertilización In Vitro , Líquido Folicular/química , Humanos , MicroARNs/análisis , MicroARNs/genética , Oogénesis/genética , Transcriptoma , Adulto Joven
19.
Ticks Tick Borne Dis ; 10(5): 1085-1095, 2019 08.
Artículo en Inglés | MEDLINE | ID: mdl-31186201

RESUMEN

The present experiment was conducted to evaluate and compare the impact of Ageratum conyzoides plant extract (ACE) with routinely used synthetic acaricides i.e., amitraz and coumaphos on the oogenesis of engorged adult females of Rhipicephalus microplus tick. On the day of dropping from the host, panoistic ovary of R. microplus appeared white in colour, horseshoe shaped, hollow tubular organ with immature oocytes predominantly in dorsal groove. Different developmental stages of oocytes (I-V) proceed simultaneously and asynchronously. Oocytes showed gradual increase in size, deep brown colored with accumulation of eggs in oviduct during 24-72 hours of development.At LC90 concentration a highly significant (p < 0.001) cessation of egg laying after exposure to amitraz and ACE while significant reduction (p < 0.01) of egg laying in coumaphos treated ticks was observed. Upon dissection of treated ticks, uterus and oviduct packed with eggs, which failed to pass out was observed. The histo-architectural alterations including presence of extensive vacuolation, alteration of oocyte morphology, deformation of chorion and disorganization of yolk granules were observed in the treated ovaries. Histochemically, low level of storage or synthesis of essential elements viz., proteins, polysaccharides and lipids in treated oocytes responsible for reduction of fertility and inhibition of progress of vitellogenesis was observed.


Asunto(s)
Acaricidas/farmacología , Ageratum/química , Cumafos/farmacología , Extractos Vegetales/farmacología , Rhipicephalus/efectos de los fármacos , Toluidinas/farmacología , Animales , Relación Dosis-Respuesta a Droga , Femenino , Oogénesis/efectos de los fármacos , Oogénesis/fisiología , Extractos Vegetales/química , Rhipicephalus/fisiología
20.
Int J Mol Sci ; 20(12)2019 Jun 12.
Artículo en Inglés | MEDLINE | ID: mdl-31212770

RESUMEN

Endocannabinoids are key-players of female fertility and potential biomarkers of reproductive dysfunctions. Here, we investigated localization and expression of cannabinoid receptor type-1 and -2 (CB1R and CB2R), G-protein coupled receptor 55 (GPR55), and transient receptor potential vanilloid type 1 channel (TRPV1) in mouse oocytes collected at different stages of in vivo meiotic maturation (germinal vesicle, GV; metaphase I, MI; metaphase II, MII) through qPCR, confocal imaging, and western blot. Despite the significant decrease in CB1R, CB2R, and GPR55 mRNAs occurring from GV to MII, CB2R and GPR55 protein contents increased during the same period. At GV, only CB1R was localized in oolemma, but it completely disappeared at MI. TRPV1 was always undetectable. When oocytes were in vitro matured with CB1R and CB2R but not GPR55 antagonists, a significant delay of GV breakdown occurred, sustained by elevated intraoocyte cAMP concentration. Although CBRs antagonists did not affect polar body I emission or chromosome alignment, GPR55 antagonist impaired in ~75% of oocytes the formation of normal-sized MI and MII spindles. These findings open a new avenue to interrogate oocyte pathophysiology and offer potentially new targets for the therapy of reproductive alterations.


Asunto(s)
Oocitos/citología , Oocitos/metabolismo , Oogénesis , Receptores de Cannabinoides/metabolismo , Animales , Antagonistas de Receptores de Cannabinoides/farmacología , Diferenciación Celular/genética , AMP Cíclico/metabolismo , Endocannabinoides/metabolismo , Expresión Génica , Ratones , Oocitos/efectos de los fármacos , Oogénesis/genética , Unión Proteica , ARN Mensajero/genética , Receptores de Cannabinoides/genética
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