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Adult T-cell leukemia/lymphoma (ATLL) is a type of cancer with aggressive and rapid progression and has been linked to infection with the human T-cell lymphotropic virus type 1 (HTLV-1). Standard treatment consists of chemotherapy,... more
Adult T-cell leukemia/lymphoma (ATLL) is a type of cancer with aggressive and rapid progression and has been linked to infection with the human T-cell lymphotropic virus type 1 (HTLV-1). Standard treatment consists of chemotherapy, radiation therapy, and stem-cell transplant, but the development of chemoresistance is common. Despite major advances in understanding the pathogenesis of the disease, patients who do not respond or achieve only transient responses remain a challenge for clinicians. Exosomes are small extracellular vesicles (30-150 nm in diameter) involved in intercellular communication and biologic material transference (including mRNA, miRNA, lipids, and metabolites) between the cells. Exosomes are an emerging key to understanding cancer cell communication and signaling pathway activation during the chemoresistance process. Our hypothesis is that exosomes secreted by chemoresistant (CR) ATLL cells can drive phenotypic changes in nonresistant ATLL cells. For this purpose...
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Background: In vitro production of bovine embryos is a well-established technology, but the in vitro culture (IVC) system still warrants improvements, especially regarding embryo quality. This study aimed to evaluate the effect of... more
Background: In vitro production of bovine embryos is a well-established technology, but the in vitro culture (IVC) system still warrants improvements, especially regarding embryo quality. This study aimed to evaluate the effect of extracellular vesicles (EVs) isolated from oviductal (OF) and uterine fluid (UF) in sequential IVC on the development and quality of bovine embryos. Zygotes were cultured in SOF supplemented with either BSA or EVs-depleted fetal calf serum (dFCS) in the presence (BSA-EV and dFCS-EV) or absence of EVs from OF (D1 to D4) and UF (D5 to D8), mimicking in vivo conditions. EVs from oviducts (early luteal phase) and uterine horns (mid-luteal phase) from slaughtered heifers were isolated by Size Exclusion Chromatography. Blastocyst rate was recorded on Days 7-8 and their quality was assessed based on lipid contents, mitochondrial activity and total cell numbers, as well as survival rate after vitrification. Relative mRNA abundance for lipid metabolism related tran...
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Supplemental data.
MicroRNAs (miRNAs) are key regulators of gene expression, potentially affecting several biological processes, whose function can be altered by sequence variation. Hence, the integration of single nucleotide polymorphisms (SNP) and miRNAs... more
MicroRNAs (miRNAs) are key regulators of gene expression, potentially affecting several biological processes, whose function can be altered by sequence variation. Hence, the integration of single nucleotide polymorphisms (SNP) and miRNAs can explain individual differences in economic traits. To provide new insights into the effects of SNPs on miRNAs and their related target genes, we carried out a multi-omic analysis to identify SNPs in miRNA mature sequences (miR-SNPs) associated with fatty acid (FA) composition in the Nelore cattle. As a result, we identified 3 miR-SNPs in different miRNAs (bta-miR-2419-3p, bta-miR-193a-2, and bta-miR-1291) significantly associated with FA traits (p-value < 0.02, Bonferroni corrected). Among these, the rs110817643C>T, located in the seed sequence of the bta-miR-1291, was associated with different ω6 FAs, polyunsaturated FA, and polyunsaturated:saturated FA ratios. Concerning the other two miR-SNPs, the rs43400521T>C (located in the bta-mi...
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Abstract The objective was to evaluate meat quality of Rubia Gallega x Nelore (RGN) crossbred compared to Nelore (N) cattle, using mRNA analysis of calpain 1 and 2, calpastatin, myostatin, and TGFβ; and protein abundance profile post... more
Abstract The objective was to evaluate meat quality of Rubia Gallega x Nelore (RGN) crossbred compared to Nelore (N) cattle, using mRNA analysis of calpain 1 and 2, calpastatin, myostatin, and TGFβ; and protein abundance profile post mortem based on aged beef of the different genetic groups. Sixteen N and 16 RGN were finished in feedlot by 120 days and harvested. RGN beef presented greater tenderness. Transcripts analysis showed increased levels of TGFβ and MSTN in RGN, indicating differences in muscle fiber formation. However, no difference was observed in mRNA for CAPN1, CAPN2 and CAST. Protein analysis demonstrated increased troponin-T degradation in RGN. Interaction for HSP 27 degradation was observed, indicating the degradation of this protein is faster and greater in RGN than N. No difference in myostatin protein abundance was found. In general, RGN cattle improves beef quality and these differences may be explained by TGFβ and MSTN gene expression, troponin-T degradation and HSP abundance.
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Oocyte maturation occurs concomitantly with oocyte chromatin condensation and transcriptional decrease. However, gene expression control can occur in these cells post-transcriptionally, by miRNA-mediated mechanisms. The miRNA expression... more
Oocyte maturation occurs concomitantly with oocyte chromatin condensation and transcriptional decrease. However, gene expression control can occur in these cells post-transcriptionally, by miRNA-mediated mechanisms. The miRNA expression profiles of bovine oocytes during early and late steps of maturation are poorly known. It is also unknown the repercussion of the exposure of bovine oocytes to in vitro culture conditions during maturation on miRNAs expression. This study aimed to determine miRNA expression in immature bovine oocytes and in mature bovine oocytes undergoing in vivo and in vitro maturation, to identify miRNAs differentially expressed among these groups and the pathways regulated by them. For that, pools (3 per group) of 20 denuded-oocytes obtained from immature and in vitro- and in vivo-matured COCs were used for miRNA extraction using a combination of TRizol reagent (Invitrogen) and the miRNeasy kit (Qiagen). Reverse transcription was carried out using the miScript PC...
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The ability to predict superstimulatory response would be a beneficial tool in assisted reproduction. Using small RNAseq technology, we profiled extracellular vesicle microRNA (EV-miRNA) abundance in the blood plasma of heifers exhibiting... more
The ability to predict superstimulatory response would be a beneficial tool in assisted reproduction. Using small RNAseq technology, we profiled extracellular vesicle microRNA (EV-miRNA) abundance in the blood plasma of heifers exhibiting variable responses to superstimulation. Estrous synchronized crossbred beef heifers (n = 25) were superstimulated and blood samples were collected from each heifer on Day 7 of consecutive unstimulated (U) and superstimulated (S) cycles. A subset of high (H) and low (L) responders was selected depending on their response to superstimulation and EV-miRNA profiles were analysed at both time-points in each heifer. Approximately 200 known miRNAs were detected in each sample with 144 commonly detected in all samples. A total of 12 and 14 miRNAs were dysregulated in UH vs. UL and in SH vs. SL heifers, respectively. Interestingly, miR-206 and miR-6517 exhibited the same differential expression pattern in H compared to L heifers both before and after supers...
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Cell communication within the ovarian follicle is crucial during folliculogenesis to assure an ideal environment for the oocyte to achieve full developmental competence. Intercellular communication is facilitated by the presence of... more
Cell communication within the ovarian follicle is crucial during folliculogenesis to assure an ideal environment for the oocyte to achieve full developmental competence. Intercellular communication is facilitated by the presence of follicular fluid, which mediates the transfer of signaling molecules. Recently, extracellular vesicles (exosomes and microvesicles) containing mRNAs, miRNAs and proteins were described in mammalian follicular fluid. Besides these molecules, extracellular vesicles (EVs) can mediate the transfer of lipids that can act as signal transducers activating second messengers and modulating intracellular pathways. Our goal was to determine the lipid profile of exosomes (small extracellular vesicles) and microvesicles (large extracellular vesicles) from bovine ovarian follicles containing oocytes with different developmental capabilities to verify potential relationships to competence. Using mass spectrometry, we examined the lipid content of EVs present in the follicular fluid of follicles enclosing oocytes that were either unable to cleave (NCLEAVE), arrested at cleavage stage (CLEAVE), or developed to the blastocyst stage (BLAST) after parthenogenetic activation. Although most of the 514 lipids identified in the follicular fluid EVs were common among all groups, 10 exosome-derived lipids and 15 microvesicle-derived lipids were present exclusively in the BLAST group, suggesting a potential relationship with developmental competence. Therefore, our data indicate that the EVs present in follicular fluid of antral follicles of similar morphology contain lipids that may be used as biomarkers associated with the developmental capability of the oocyte to develop to the blastocyst stage.
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Preterm labor (PTL) and Preterm Premature Rupture of Membranes (PPROM) impose substantial morbimortality on mothers and newborns. Exosomes act in intercellular communication carrying molecules involved in physiopathological processes.... more
Preterm labor (PTL) and Preterm Premature Rupture of Membranes (PPROM) impose substantial morbimortality on mothers and newborns. Exosomes act in intercellular communication carrying molecules involved in physiopathological processes. Little is known about exosomal proteins in prematurity. Our aim was to evaluate the protein expression of hemopexin, C1 inhibitor (C1INH) and alpha-2-macroglobulin (A2M) from circulating exosomes of women with PTL and PPROM. Plasma was obtained from PTL, PPROM, Term in labor and Term out of labor (T) patients, exosomes were isolated by ultracentrifugation, then lysed and the proteins quantified. Western Blot (WB) and Nanoparticle Tracking Analysis (NTA) were performed. Data were compared by Kruskal–Wallis, unpaired T-test and one-way ANOVA. WB and NTA confirmed exosome isolation (concentration: 4.3 × 1010 particles/ml ± 1.9 × 1010). There was no difference regarding hemopexin or C1INH expression between the groups. For A2M, the fold change was signific...
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Lymphoma is the most common type of canine hematological malignancy where the multicentric (cMCL) form accounts for 75% of all cases. The standard treatment is the CHOP chemotherapy protocols that include cyclophosphamide, doxorubicin,... more
Lymphoma is the most common type of canine hematological malignancy where the multicentric (cMCL) form accounts for 75% of all cases. The standard treatment is the CHOP chemotherapy protocols that include cyclophosphamide, doxorubicin, vincristine and prednisone, where the majority of dogs achieve complete/partial response; however, it is very important to predict non-responsive cases to improve treatment and to develop new targeted therapies. Here we evaluate a liquid biopsy approach based on serum Small Extracellular Vesicles enriched for exosomes (SEVs) to predict cMCL chemotherapy response. Nineteen dogs at the end of the 19-week chemotherapy protocol (8 Complete Response and 11 Progressive Disease) were evaluated for serum SEVs size, concentration and screened for 95 oncomirs. PD patients had higher SEVs concentration at the diagnosis than CR patients (P = 0.034). The ROC curve was significant for SEVs concentration to predict the response to CHOP (AUC = 0.8011, P = 0.0287). A ...
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Extracellular vesicles (EVs) are nanoparticles secreted by ovarian follicle cells. Extracellular vesicles are an important form of intercellular communication, since they carry bioactive contents, such as microRNAs (miRNAs), mRNAs, and... more
Extracellular vesicles (EVs) are nanoparticles secreted by ovarian follicle cells. Extracellular vesicles are an important form of intercellular communication, since they carry bioactive contents, such as microRNAs (miRNAs), mRNAs, and proteins. MicroRNAs are small noncoding RNA capable of modulating mRNA translation. Thus, EVs can play a role in follicle and oocyte development. However, it is not clear if EV contents vary with the estrous cycle stage. The aim of this study was to investigate the bovine miRNA content in EVs obtained from follicles at different estrous cycle stages, which are associated with different progesterone (P4) levels in the follicular fluid (FF). We collected FF from 3 to 6 mm follicles and evaluated the miRNA profile of the EVs and their effects on cumulus-oocyte complexes during in vitro maturation. We observed that EVs from low P4 group have a higher abundance of miRNAs predicted to modulate pathways, such as MAPK, RNA transport, Hippo, Cell cycle, FoxO, ...
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Introduction. Pluripotent stem cells are believed to have greater clinical potential than mesenchymal stem cells due to their ability to differentiate into almost any cell type of an organism, and since 2006, the generation of... more
Introduction. Pluripotent stem cells are believed to have greater clinical potential than mesenchymal stem cells due to their ability to differentiate into almost any cell type of an organism, and since 2006, the generation of patient-specific induced pluripotent stem cells (iPSCs) has become possible in multiple species. Objectives. We hypothesize that different cell types respond differently to the reprogramming process; thus, the goals of this study were to isolate and characterize equine adult and fetal cells and induce these cells to pluripotency for future regenerative and translational purposes. Methods. Adult equine fibroblasts (eFibros) and mesenchymal cells derived from the bone marrow (eBMmsc), adipose tissue (eADmsc), and umbilical cord tissue (eUCmsc) were isolated, their multipotency was characterized, and the cells were induced in vitro into pluripotency (eiPSCs). eiPSCs were generated through a lentiviral system using the factors OCT4, SOX2, c-MYC, and KLF4. The morp...
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Semen fertilizing potential is dependent upon the morphological, functional and molecular attributes of sperm. Sperm microRNAs (miRNAs) were recently shown to hold promise regarding their association with different fertility phenotypes.... more
Semen fertilizing potential is dependent upon the morphological, functional and molecular attributes of sperm. Sperm microRNAs (miRNAs) were recently shown to hold promise regarding their association with different fertility phenotypes. However, their role in fertility regulation remains to be determined. We postulated that sperm miRNAs might regulate early embryonic development. From this perspective, sperm quality and 380 sperm miRNAs were investigated in frozen–thawed semen from high (HF; 54.3 ± 1.0% pregnancy rate) and low (LF; 41.5 ± 2.3%) fertility bulls. Out of nine miRNAs that showed different levels in sperm cells, miR-216b was present at lower levels in HF sperm cells and zygotes. Among miR-216b target genes (K-RAS, BECN1 and JUN), K-RAS, related to cell proliferation, revealed a higher level in HF two-cell embryos. First cleavage rate, blastocyst cell number and division number were also higher in HF. In addition, by using a model based on polyspermy embryos, we demonstra...
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Human T-cell lymphotropic virus type 1 (HTLV-1) is the etiological agent of HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP) and adult T-cell leukemia/lymphoma (ATLL). The development of HAM/TSP, a chronic... more
Human T-cell lymphotropic virus type 1 (HTLV-1) is the etiological agent of HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP) and adult T-cell leukemia/lymphoma (ATLL). The development of HAM/TSP, a chronic neuroinflammatory disease, is correlated to complex interaction between the host immune response and the infecting virus. Tax expression plays important role in HAM/TSP pathogenesis by activating various cellular genes, including the cytokines IFN-γ and TNF-α. Exosomes have emerged as an important factor of cell-to-cell communication contributing to diverse cellular processes, including immune modulation. Considering the potential role of exosomes in modulating the immune response and inflammation, the main objective of this study was to examine if HTLV-1-infected cells produce exosomes carrying viral proteins or inflammatory molecules, which can participate in the chronic inflammation which is observed in patients with HAM/TSP. Exosomes were isolated from HTLV-...
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Early mammalian embryos derived from in vitro fertilization are exposed to conditions distinct from the native oviduct-uterine environment, including atmospheric oxygen that promotes cellular oxidative stress and alters gene expression.... more
Early mammalian embryos derived from in vitro fertilization are exposed to conditions distinct from the native oviduct-uterine environment, including atmospheric oxygen that promotes cellular oxidative stress and alters gene expression. High oxygen partial pressure during embryo development is associated with low pregnancy rates and increased embryonic apoptosis. We investigated how bovine embryos responded to high (20% O2 ) or low (5% O2 ) oxygen partial pressure during in vitro culture, evaluating levels of reactive oxygen species (ROS) as well as changes in the expression of oxidative stress- and epigenetic-related transcripts and miRNAs in blastocysts. Additionally, we determined the global DNA methylation levels in the resulting embryos. Our results indicated that bovine blastocysts produced in vitro under high oxygen partial pressure possessed elevated ROS abundance and exhibited increased expression of CAT, GLRX2, KEAP1, NFR2, PRDX1, PRDX3, SOD1, TXN, and TXNRD1, versus reduc...
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Cell-secreted vesicles called microvesicles (MVs) and exosomes (EXOs) are thought to mediate cell communication, and are present in different body fluids. MVs (100-1000 nm) and EXOs (30-150 nm) are secreted by cells using different... more
Cell-secreted vesicles called microvesicles (MVs) and exosomes (EXOs) are thought to mediate cell communication, and are present in different body fluids. MVs (100-1000 nm) and EXOs (30-150 nm) are secreted by cells using different cellular pathways and contain similar bioactive material such as mRNAs, miRNAs, and proteins. Recently, these vesicles have been identified in ovarian follicular fluid of mares, cows and woman. The presence of these vesicles in ovarian follicular fluid presents an opportunity for the discovery of novel diagnostic markers, as well as therapeutic tools to improve fertility. Cows are good models to study follicular cell communication due to the easy access to samples and well-established protocols for artificial reproductive techniques. In order to study and characterize cell-secreted vesicles in bovine follicular fluid, we isolated MVs and EXOs from immature bovine follicles and determined purity of the preparations based on morphology and RNA/protein conte...
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Ovarian follicular development is a controlled series of events culminating with an ovulatory or atretic follicle. MicroRNAs (miRNAs) are small noncoding RNAs involved in translational regulation of genes in different developmental... more
Ovarian follicular development is a controlled series of events culminating with an ovulatory or atretic follicle. MicroRNAs (miRNAs) are small noncoding RNAs involved in translational regulation of genes in different developmental processes. Deletion of Dicer in mice ovaries demonstrated the importance of miRNAs in reproduction, which led to infertility. The miRNAs were thought to act only within host cells; however, these molecules are also present in cell-secreted vesicles. These vesicles are present in body fluids such as milk, serum, and ovarian follicular fluid. Vesicles are secreted in extracellular fluids and travel from donor to target cells, mediating transfer of bioactive material. Herein we discuss the role of hormonal-regulated miRNAs within different ovarian follicular cells as well as cell-secreted vesicles participation in mammalian ovarian follicular fluid. Furthermore, we discuss the possibility of miRNAs transference mediated by cell-secreted vesicles present in o...
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Fertility prediction is performed evaluating several sperm morphological and functional aspects (MFF). In general, MFF features present high correlation with fertility. However, there are ejaculates that, although present normal MFF... more
Fertility prediction is performed evaluating several sperm morphological and functional aspects (MFF). In general, MFF features present high correlation with fertility. However, there are ejaculates that, although present normal MFF features, have poor fertility results. So, there is necessity to investigate potential fertility molecular markers which are able to predict fertility rates. Among them are the microRNAs (miRNAs), which are post-transcriptional regulators molecules that are important to spermatogenesis, sperm maturation and also embryo development. Besides, miRNAs are related with men and bull fertility and are highly regulated on masculine tract. Currently, there are around 790 mature miRNA sequences in bovine, which could be used as biomarkers. Nevertheless, the big challenge is to show how miRNAs regulate fertility and their functions on sperm and embryo development. Thus, the goal of this review is to compile the data of scientific literature about miRNAs and male fe...
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Information on molecular mechanisms through which sex-steroids regulate oviductal function to support early embryo development is lacking. Here, we hypothesized that the periovulatory endocrine milieu affects the miRNA processing... more
Information on molecular mechanisms through which sex-steroids regulate oviductal function to support early embryo development is lacking. Here, we hypothesized that the periovulatory endocrine milieu affects the miRNA processing machinery and miRNA expression in bovine oviductal tissues. Growth of the preovulatory follicle was controlled to obtain cows that ovulated a small follicle (SF) and subsequently bore a small corpus luteum (CL; SF-SCL) or a large follicle (LF) and large CL (LF-LCL). These groups differed in the periovulatory plasmatic sex-steroid’s concentrations. Ampulla and isthmus samples were collected on day four of the estrous cycle. Abundance of DROSHA, DICER1, and AGO4 transcripts was greater in the ampulla than the isthmus. In the ampulla, transcription of these genes was greater for the SF-SCL group, while the opposite was observed in the isthmus. The expression of the 88 most abundant miRNAs and 14 miRNAs in the ampulla and 34 miRNAs in isthmus were differentiall...
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Early embryonic development occurs in the oviduct, where an ideal microenvironment is provided by the epithelial cells and by the oviductal fluid produced by these cells. The oviductal fluid contains small extracellular vesicles (sEVs),... more
Early embryonic development occurs in the oviduct, where an ideal microenvironment is provided by the epithelial cells and by the oviductal fluid produced by these cells. The oviductal fluid contains small extracellular vesicles (sEVs), which through their contents, including microRNAs (miRNAs), can ensure proper cell communication between the mother and the embryo. However, little is known about the modulation of miRNAs within oviductal epithelial cells (OECs) and sEVs from the oviductal fluid in pregnant cows. In this study, we evaluate the miRNAs profile in sEVs from the oviductal flushing (OF-sEVs) and OECs from pregnant cows compared to non-pregnant, at 120 h after ovulation induction. In OF-sEVs, eight miRNAs (bta-miR-126-5p, bta-miR-129, bta-miR-140, bta-miR-188, bta-miR-219, bta-miR-345-3p, bta-miR-4523, and bta-miR-760-3p) were up-regulated in pregnant and one miRNA (bta-miR-331-5p) was up-regulated in non-pregnant cows. In OECs, six miRNAs (bta-miR-133b, bta-miR-205, bta-m...
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We evaluated the effect of the antral follicle count (AFC) on ovarian follicular dynamics, pregnancy rates, progesterone concentrations, and transcriptional patterns of genes in Nelore cattle (Bos taurus indicus) after a timed artificial... more
We evaluated the effect of the antral follicle count (AFC) on ovarian follicular dynamics, pregnancy rates, progesterone concentrations, and transcriptional patterns of genes in Nelore cattle (Bos taurus indicus) after a timed artificial insemination (TAI) programme. Cows were separated based on the AFC, and those with a high AFC showed a larger (P < 0.0001) ovarian diameter and area than those with a very low AFC. Females with a very low AFC exhibited a larger (P < 0.01) diameter of the dominant follicle at TAI (13.6 ± 0.3 vs. 12.2 ± 0.4 mm) and a tendency (P = 0.06) to have different serum progesterone concentrations (2.9 ± 0.3 vs. 2.1 ± 0.3 ng/mL; on day 18, considering day 0 as the beginning of the synchronization protocol) than those with a high AFC. The pregnancy rate was higher (P ≤ 0.05) in animals with a very low (57.9%) and low (53.1%) AFC than in those with a high AFC (45.2%). The expression of genes related to intercellular communication, meiotic control, epigeneti...
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Embryo–maternal crosstalk is an important event that involves many biological processes, which must occur perfectly for pregnancy success. This complex communication starts from the zygote stage within the oviduct and continues in the... more
Embryo–maternal crosstalk is an important event that involves many biological processes, which must occur perfectly for pregnancy success. This complex communication starts from the zygote stage within the oviduct and continues in the uterus up to the end of pregnancy. Small extracellular vesicles (EVs) are part of this communication and carry bioactive molecules such as proteins, lipids, mRNA, and miRNA. Small EVs are present in the oviductal and uterine fluid and have important functions during fertilization and early embryonic development. Embryonic cells are able to uptake oviductal and endometrium-derived small EVs. Conversely, embryo-derived EVs might modulate oviductal and uterine function. In this review, our aim is to demonstrate the role of extracellular vesicles modulating embryo–maternal interactions during early pregnancy.
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Orchestrated events, including extensive changes in epigenetic marks, allow a somatic nucleus to become totipotent after transfer into an oocyte, a process termed nuclear reprogramming. Recently, several strategies have been applied in... more
Orchestrated events, including extensive changes in epigenetic marks, allow a somatic nucleus to become totipotent after transfer into an oocyte, a process termed nuclear reprogramming. Recently, several strategies have been applied in order to improve reprogramming efficiency, mainly focused on removing repressive epigenetic marks such as histone methylation from the somatic nucleus. Herein we used the specific and non-toxic chemical probe UNC0638 to inhibit the catalytic activity of the histone metyltransferases EHMT1 and EHMT2. Either the donor cell (before reconstruction) or the early embryo was exposed to the probe to assess its effect on developmental rates and epigenetic marks. First, we showed that the treatment of bovine fibroblasts with UNC0638 did mitigate the levels of H3K9me2. Moreover, H3K9me2 levels were decreased in cloned embryos regardless of treating either donor cells or early embryos with UNC0638. Additional epigenetic marks such as H3K9me3, 5mC, and 5hmC were a...
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Mammalian ovarian follicular growth is characterized by development of a large fluid filled antrum that separates mural granulosa cells and cumulus cells. Extensive communication between the different cell types is necessary for... more
Mammalian ovarian follicular growth is characterized by development of a large fluid filled antrum that separates mural granulosa cells and cumulus cells. Extensive communication between the different cell types is necessary for maturation of a developmentally competent oocyte. Here, we describe an approach for the isolation of cell-secreted exosomes from ovarian follicular fluid, identification of small RNAs (i.e., microRNAs) in exosomes, labeling of exosomes, and examining cell uptake of exosomes by follicular cells.
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The influence of in vitro maturation (IVM) in oocytes is still not totally understood. The aim of this study was to determine the influence of IVM on the metabolism and homeostasis of bovine cumulus-oocyte complexes. In the present study,... more
The influence of in vitro maturation (IVM) in oocytes is still not totally understood. The aim of this study was to determine the influence of IVM on the metabolism and homeostasis of bovine cumulus-oocyte complexes. In the present study, we demonstrated that IVM leads to accumulation of neutral lipids associated with differential levels of the mono-, di-, and tri-acylglycerols in both cumulus cells and oocytes. We observed that in vitro-matured oocytes exhibited decreased glutathione and reactive oxygen species levels and a lower ATP/ADP ratio when compared to in vivo-matured oocytes, with no significant differences in metabolism and stress related mRNA or miRNA levels. Moreover, in addition to an increase in lipids in in vitro-matured cumulus cells, fatty acid synthesis and accumulation as well as glycolysis pathway genes were upregulated, whereas those affiliated with the -oxidation pathway were decreased. Our gene expression data in cumulus cells suggest the disruption of endopl...