Serum microRNAs in buffalo cows: Potential biomarkers of pregnancy (original) (raw)
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Frontiers in Cell and Developmental Biology, 2021
Precise early pregnancy diagnosis in dairy animals is of utmost importance for an efficient dairy production system. Not detecting a dairy animal pregnant sufficiently early after the breeding results to extending the unproductive time of their milk production cycle and causes substantial economic loss for a dairy producer. At present, the most conventional and authentic pregnancy confirmation practice in cows and buffaloes is rectal palpation of the reproductive organs at Days 35–40 after insemination, which sometime leads to considering an animal as false pregnant. Other alternative methods available for early pregnancy diagnosis lack either accuracy or reproducibility or require elaborate instrumentation and laboratory setup not feasible to practice at farmers’ doorstep. The present study was aimed at establishment of the microRNA (miRNA) repertoire of the placentome in buffaloes, which could capture the event of the cross talk between a growing embryo and a dam, through fetal co...
Circulating miRNAs in maternal plasma as potential biomarkers of early pregnancy in sheep
Frontiers in Genetics
MicroRNA (miRNA) plays an important role in the control of gene expression and is implied in many biological functions, including embryo implantation and development. The aim was to assess plasma miRNA profiles during the peri-implantation and ascertain potential candidate miRNA markers for early pregnancy diagnosis in ovine plasma. The plasma samples were obtained from a total of 24 ewes on days 12 (pre-implantation; P12, n = 4), 16 (implantation; P16, n = 4) and 22 (post-implantation; P22, n = 4) after mating, and on their corresponding days of 12 (Pre-C; C12, n = 4), 16 (Imp-C; C16, n = 4) and 22 (Post-C; C22, n = 4) of the estrous cycle. The miRNA profiles in plasma were assessed by microarray technology. We detected the presence of 60 ovine-specific miRNAs in plasma samples. Of these miRNAs, 22 demonstrated a differential expression pattern, especially between the estrous cycle and early pregnancy, and targeted 521 genes. Two miRNAs (oar-miR-218a and oar-miR-1185-3p) were confi...
The microRNAs important for ovarian and early embryonic development in cattle
Recent progress in genomic analysis and other biochemical methods has led to the discovery of a large population of microRNAs (miRNAs), which have been demonstrated to play important roles in diseases and a wide range of developmental processes. Characterization of miRNA expression profiles in different stages of ovarian follicular development and early embryogenesis has suggested the potential roles of miRNAs in follicular development, maturation of oocytes, and preimplantation embryonic development. This review focuses on the current studies of miRNAs involved in ovarian and early embryonic development in cattle.
Analysis of immune-related microRNAs in cows and newborn calves
Journal of Dairy Science, 2023
Bovine colostrum contains a high concentration of immune-related microRNAs (miRNAs) that are packaged in exosomes and are very stable. In this study, 5 immune-related miRNAs (miR-142-5p, miR-150, miR-155, miR-181a, and miR-223) were quantified in dam blood, colostrum, and calf blood using reverse transcription quantitative PCR. Their levels in calf blood after colostrum ingestion were investigated to assess whether miRNAs are transferred from the dam to newborn calves. Three groups of Holstein-Friesian bull calves were bottle-fed 2 L of colostrum or milk from different sources twice per day. The group A calves received colostrum from their own dam and the group B calves were fed foster dam colostrum. Each pair of group A and group B calves were fed identical colostrum from the same milking of the corresponding group A dam for 3 d and then bulk tank milk for 7 d after birth. Group C calves were fed only 2L of "pooled colostrum" from multiple dams d 0 to 4 postpartum, and then fed bulk tank milk thereafter for 7 d after birth. The groups were fed colostrum from different sources and different amounts to assess possible miRNA absorption from the colostrum. All miRNAs were at the highest level in colostrum at d 0 and then decreased rapidly after d 1. The level of miR-150 had the largest decrease from 489 × 106 copies/µL (d 0) to 78 × 106 copies/ µL (d 1). MicroRNA-223 and miR-155 were the most abundant in both colostrum and milk. Dam colostrum had significantly higher levels of miR-142-5p, miR-155, and miR-181a than the bulk tank milk. However, only the miR-155 concentration was significantly higher in the dam colostrum than in the pooled colostrum. The concentrations of miRNAs in the colostrum were less than in the cow blood (100-to 1,000-fold less). There was no significant correlation between the level of miR-NAs in the dam blood and their colostrum, suggesting that miRNA is synthesized locally by the mammary gland rather than being transferred from the blood. MicroRNA-223 had the highest level in both calf and cow blood compared with the other 4 immune-related miRNAs. Calves were born with high levels of immunerelated miRNAs in their blood, and there were no significant differences in miRNA levels between the 3 calf groups at birth or after they were fed different colostrum. This suggests that these miRNAs were not transferred from the colostrum to the newborn calves.
Scientific Reports, 2022
In buffalo (Bubalus bubalis) reproductive seasonality, causing cycles of milk production, is one of the major factors affecting farming profitability. Follicular fluid (FF) contains extracellular vesicles (EVs) playing an important role in modulating oocyte developmental competence and carrying microRNAs (miRNAs) essential for in vitro fertilization outcomes. The aim of this work was to characterize the FF-EVs-miRNA cargo of antral (An) and preovulatory (pO) follicles collected in the breeding (BS) and non-breeding (NBS) seasons, to unravel the molecular causes of the reduced oocyte competence recorded in buffalo during the NBS. In total, 1335 miRNAs (538 known Bos taurus miRNAs, 324 homologous to known miRNAs from other species and 473 new candidate miRNAs) were found. We identified 413 differentially expressed miRNAs (DE-miRNAs) (FDR < 0.05) between An and pO groups. A subset of the most significant DE-miRNAs between An and pO groups targets genes which function is related to the lipid and steroid metabolism, response to glucocorticoid and oestradiol stimulus. Comparison between BS and NBS showed 14 and 12 DE-miRNAs in An-FF-EVs and pO-FF-EVs, which regulate IL6 release and cellular adhesion, respectively. In conclusion, these results demonstrated that the miRNA cargo of buffalo FF-EVs varies in relation to both follicular development and season. The importance of buffalo (Bubalus bubalis) breeding is clearly indicated by the positive growth trend all over the world (Faostat.fao.org/faostat), due to specific features making this species a valuable protein source, particularly for tropical countries. A peculiar situation is described in Italy, where the success of buffalo breeding is closely related to the production of mozzarella cheese, highly requested around the world. A major limiting factor is reproductive seasonality, impeding continuity of milk production throughout the year, and hence not allowing to meet the market demand. Buffalo is a short-day breeder, with an increased reproductive activity observed during decreasing day length months 1,2. In Italy, where the Italian Mediterranean buffalo breed has been selected, the seasonality pattern shows an opposite trend to the market request. Therefore, the out of breeding mating strategy (OBMS), has been efficiently applied to distribute calving more evenly during the year 2. However, forcing buffalo cows to conceive during the non-breeding season (NBS) may lead to extended post-partum anestrus, higher rates of embryonic mortality and overall reduced fertility 1,3,4. In previous studies it was demonstrated that embryonic mortality is in part caused by impaired luteal function, and consequently reduced progesterone secretion 5. This in turn interferes with embryo growth that is accompanied by transcriptomic and proteomic changes at the level of embryos and chorioamnios/caruncles 6,7 , definitively hampering embryo attachment. Embryonic mortality during the NBS is also in part due to reduced oocyte developmental competence, as shown by the decreased cleavage and blastocyst rates obtained after in vitro fertilization during increasing daylight months in Italian Mediterranean buffaloes 8,9. A seasonal effect on follicular population and embryo outcomes was also reported by other authors 10,11. A poorer oocyte quality was observed in Murrah buffalo heifers during
Expression of miRNAs in Bull Spermatozoa Correlates with Fertility Rates
Bull spermatozoa are rich in active miRNAs, and it has been shown that specific spermborne miRNAs can be linked to fertility. Thus, expression profiling of spermatozoa could be helpful for understanding male fertility and the ability of spermatozoa to initiate and sustain zygotic, embryonic and foetal development. Herein we hypothesized that bulls with moderate to high fertility can be identified by differences in amounts of certain miRNAs between their ejaculates. RNA samples from spermatozoa of eight brother pairs (one bull with high and one with moderate NRR in each pair) of the Holstein breed were prepared. miRNA was isolated, and the expression of 178 miRNAs was determined by RT-qPCR. Important findings were that highly expressed miRNAs, not linked toNRR status, were identified in the bull sperm samples, which indicate that these miRNAs have an important role in early embryogenesis. A large fraction of the targets genes were phosphoproteins and genes involved in the regulation of transcription. Seven miRNAs (mir-502-5p, mir-1249, mir-320a, mir-34c-3p, mir-19b-3p, mir-27a-5p and mir-148b-3p) were differentially expressed between bulls with moderate and high NRR with a strong tendency towards a higher expression ofmiRNAs in bulls with moderate fertility. Thus, bulls with a moderate NRR negatively regulate the expression of protein-coding genes, which leads to problems during the pregnancy.
Scientific Reports
Season clearly influences oocyte competence in buffalo (Bubalus bubalis); however, changes in the oocyte molecular status in relation to season are poorly understood. This study characterizes the microRNA (miRNA) and transcriptomic profiles of oocytes (OOs) and corresponding follicular cells (FCs) from buffalo ovaries collected in the breeding (BS) and non-breeding (NBS) seasons. In the BS, cleavage and blastocyst rates are significantly higher compared to NBS. Thirteen miRNAs and two mRNAs showed differential expression (DE) in FCs between BS and NBS. DE-miRNAs target gene analysis uncovered pathways associated with transforming growth factor β (TGFβ) and circadian clock photoperiod. Oocytes cluster in function of season for their miRNA content, showing 13 DE-miRNAs between BS and NBS. Between the two seasons, 22 differentially expressed genes were also observed. Gene Ontology (GO) analysis of miRNA target genes and differentially expressed genes (DEGs) in OOs highlights pathways related to triglyceride and sterol biosynthesis and storage. Co-expression analysis of miRNAs and mRNAs revealed a positive correlation between miR-296-3p and genes related to metabolism and hormone regulation. In conclusion, season significantly affects female fertility in buffalo and impacts on oocyte transcriptomic of genes related to folliculogenesis and acquisition of oocyte competence. Water buffalo (Bubalus bubalis) is an important livestock resource for both developing and developed countries. The major factor affecting buffalo farming profitability is reproductive seasonality, resulting in cycles of calving and milk production. Buffalo is a short-day breeder, with increased fertility in response to decreasing day length 1,2. This photoperiod-dependent seasonality pattern is more pronounced as distance from the equator, together with variations in the light/dark ratio, increases. In Italy, in order to satisfy market demand, out of breeding mating strategy (OBMS), consisting in interrupting sexual promiscuity or the use of artificial insemination (AI) during the breeding season (BS), is commonly utilized 2. The OBMS improves the distribution of calving throughout the year, but it reduces fertility 3. Longer post-partum anoestrus periods as well as higher incidence of embryonic mortality are observed in months with increasing daylight length and particularly in midwinter , which coincides with the transition to seasonal anoestrus at Italian latitudes 1,4. The embryonic mortality is due to inadequate luteal growth and function, resulting in reduced progesterone secretion 5. This has a negative impact on embryo growth, associated with alterations in transcriptomic and proteomic profiles of the embryos and chorioamnios/caruncles 6,7 , which ultimately impair embryo attachment to the uterine endometrium.
Changes in Oviductal Cells and Small Extracellular Vesicles miRNAs in Pregnant Cows
Frontiers in Veterinary Science, 2021
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...
†Sally Ibrahim, 2022
Background: No doubt that the corpus luteum (CL) plays a vital role in the regulation of female cyclicity in mammals. The scenarios among microRNAs (miRNAs) and their target genes and steroid hormones {estradiol (E2) and progesterone (P4)} are required for better understanding the molecular regulation of CL during its formation, maturation, and regression. We aimed to (I) study the changes in the relative abundance of miR-205, miR-26a-5p, miR-17-5p, and let-7b-5p and their target genes: LHCGR, CASP3, PCNA, AMH, and PLA2G3, during different stages of corpus luteum in Egyptian buffaloes, and (II) and to address different scenarios between steroid concentrations in the serum and the expression pattern of selected miRNAs and their targets. Methods: The paired ovaries and blood samples were collected from apparently healthy 50 buffalo cows at a private abattoir. The ovaries bearing CL were macroscopically divided according to their morphological structure and color into hemorrhagic (CLH), developing (CLD), mature (CLM), regressed (CLR), and albicans (CLA). Small pieces from different stages of CL (CLH, CLD, CLM, CLR, and CLA) were cut and immediately kept at − 80 °C for total RNA isolation and qRT-PCR. The serum was separated for steroid level estimation. Results: The LHCGR was expressed during different stages of CL, and the peak of expression was at the mid-luteal stage. The CASP3 revealed a stage-specific response at different stages of CL. The PCNA has an essential role in cellular proliferation in buffaloes CL. Both expression patterns of PLA2G3 and AMH were found over the various developmental and regression stages. It was noticed that miR-205 is conserved to target LHCGR and CASP3 transcripts. Moreover, CASP3 and AMH were targeted via miR-26a-5p. Additionally, the CASP3 and PLA2G3 were targeted via let-7b-5p. The P4 level reached its peak during CLM. There were positive and negative strong correlations between miRNAs (miR-26a-5p and miR-205), target genes (LHCGR and CASP3) during different stages of CL, and steroid hormones in the serum. Conclusions: Taken together, the orchestrated pattern among miRNAs, target genes, and steroid hormones is essential for maintaining the proper development and function of CL in buffalo cows.
2021
Despite the success of cloning technology in production of offspring across several species, its application on a wide scale is severely limited by the very low offspring rate obtained with cloned embryos. MicroRNAs (miRNAs) expression profile in cloned embryos throughout the embryonic development are reported to deviate from regular pattern. In present study, aimed at determining the dynamics of global expression of miRNA profile in cloned and IVF pre-implantation embryos at different developmental stages i.e 2-cell, 8-cell and blastocyst using next-generation sequencing. The results of this study suggested that there is a profound difference in the global miRNA profile between cloned and IVF embryos. These differences are manifested throughout the course of embryonic development. The cloned embryos differ from their IVF counterparts in the enriched GO terms of Biological Process, Molecular Function, Cellular Component and Protein Class categories in terms of the targets of differe...