Extracellular vesicle-coupled miRNA profiles in follicular fluid of cows with divergent post-calving metabolic status (original) (raw)
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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...
Abundantly Present miRNAs in Milk-Derived Extracellular Vesicles Are Conserved Between Mammals
Frontiers in Nutrition
Mammalian milk is not only a source of nutrition for the newborn, but also contains various components that regulate further development. For instance, milk is an abundant source of microRNAs (miRNAs), which are evolutionary conserved small non-coding RNAs that are involved in post-transcriptional regulation of target mRNA. MiRNAs present in milk can occur in extracellular vesicles (EVs), which are nanosized membrane vesicles released by many cell types as a means of intercellular communication. The membrane of EVs protects enclosed miRNAs from degradation and harbors molecules that allow specific targeting to recipient cells. Although several studies have investigated the miRNA content in milk EVs from individual species, little is known about the evolutionary conserved nature of EV-associated miRNAs among different species. In this study, we profiled the miRNA content of purified EVs from human and porcine milk. These data were compared to published studies on EVs from human, cow, porcine, and panda milk to assess the overlap in the top 20 most abundant miRNAs. Interestingly, several abundant miRNAs were shared between species (e.g., let-7 family members let-7a, let-7b, let-7f, and miR-148a). Moreover, these miRNAs have been implicated in immune-related functions and regulation of cell growth and signal transduction. The conservation of these miRNA among species, not only in their sequence homology, but also in their incorporation in milk EVs of several species, suggests that they are evolutionarily selected to regulate cell function in the newborn.
Scientific Reports
In dairy cows, the period from the end of lactation through the dry period and into the transition period, requires vast physiological and immunological changes critical to mammary health. The dry period is important to the success of the next lactation and intramammary infections during the dry period will adversely alter mammary function, health and milk production for the subsequent lactation. MicroRNAs (miRNAs) are small non-coding RNAs that can post transcriptionally regulate gene expression. We sought to characterize the miRNA profile in dry secretions from the last day of lactation to 3, 10, and 21 days post dry-off. We identified 816 known and 80 novel miRNAs. We found 46 miRNAs whose expression significantly changed (q-value
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.
MicroRNA expression patterns in the bovine mammary gland are affected by stage of lactation
Journal of dairy science, 2012
The objective of this work was to determine the expression pattern of microRNA (miR) associated with cellular proliferation, lipid metabolism, and innate immunity in dairy cow mammary gland tissue at different stages of lactation. The expression of miR-10a, miR-15b, miR-16, miR-21, miR-31, miR-33b, miR-145, miR-146b, miR-155, miR-181a, miR-205, miR-221, and miR-223 was studied by real-time reverse-transcription PCR in tissue (n = 7/stage) harvested via repeated biopsies during the dry period (−30 d prepartum), the fresh period (7 d postpartum), and early lactation (30 d postpartum). Except for miR-31, all miR studied increased in expression between the dry and fresh periods. Among those upregulated, the expression of miR-221 increased further at early lactation, suggesting a role in the control of endothelial cell proliferation or angiogenesis, whereas the expression of miR-223 decreased at early lactation but to a level that was greater than in the dry period, suggesting it could play a role in the mammary response to pathogens soon after parturition. The expression of miR-31, a hormonally regulated miR that inhibits cyclin gene expression, was greater at early lactation compared with the dry period. From a metabolic standpoint, the consistent upregulation of miR-33b during early lactation compared with the dry period suggests that this miR may exert some control over lipogenesis in mammary tissue. Overall, results indicate that expression of miR associated with transcriptional regulation of genes across diverse biological functions is altered by stage of lactation. The specific roles of these miR during lactation will require further research.
Serum microRNAs in buffalo cows: Potential biomarkers of pregnancy
Research in veterinary science, 2017
MiRNAs (microRNA) constitute a large family of single-stranded, non-coding small RNAs. Although the functions and target genes of most miRNAs are still unknown, it has been well established that they are involved in embryogenesis, organogenesis and neonatal birth. In recent decades, interest in buffalo breeding has largely increased worldwide thus focusing the attention on this species as a dairy purpose animal. Problems related to long calving interval, late puberty and seasonal anestrus hamper reproductive efficiency in this species. Early pregnancy diagnosis is important to shorten the calving interval and increase lifetime production on dairy animals. MicroRNAs have recently emerged as key molecules in fertilization of several species even though in buffalo, few previous studies have investigated miRNAs. The aim of this research was to identify the best miRNA reference in serum among miR-191, miR-25-3p, SNORD44, and SNORD48. Consequently, assess the expression levels of miR-103,...
Human milk (HM) contains regulatory biomolecules including miRNAs, the origin and functional significance of which are still undetermined. We used TaqMan OpenArrays to profile 681 mature miRNAs in HM cells and fat, and compared them with maternal peripheral blood mononuclear cells (PBMCs) and plasma, and bovine and soy infant formulae. HM cells and PBMCs (292 and 345 miRNAs, respectively) had higher miRNA content than HM fat and plasma (242 and 219 miRNAs, respectively) (p < 0.05). A strong association in miRNA profiles was found between HM cells and fat, whilst PBMCs and plasma were distinctly different to HM, displaying marked inter-individual variation. Considering the dominance of epithelial cells in mature milk of healthy women, these results suggest that HM miRNAs primarily originate from the mammary epithelium, whilst the maternal circulation may have a smaller contribution. Our findings demonstrate that unlike infant formulae, which contained very few human miRNA, HM is a rich source of lactation-specific miRNA, which could be used as biomarkers of the performance and health status of the lactating mammary gland. Given the recently identified stability, uptake and functionality of food-and milk-derived miRNA in vivo, HM miRNA are likely to contribute to infant protection and development. Human milk (HM) is the optimal nutrition for term infants 1. In addition to being a food source, HM confers developmental programming to the infant and protection against infections, resulting in decreased risk of sudden infant death syndrome and reduced mortality and morbidity both in the short-and long-term 2–6. These effects are mediated by HM-specific regulatory factors including both cellular and biochemical components 7–10. In contrast , artificial infant formula cannot confer such protective and developmental functions as it lacks important HM components with bioactivity 11,12. An additional unique bioactive component of HM that has been recently discovered is miRNAs 13,14. miRNAs are small non-coding RNAs, which regulate gene expression, thus control protein synthesis at the post-transcriptional level in eukaryotic cells 15. They have been identified as key regulators of diverse biological and developmental processes in eukaryotes (cell proliferation and differentiation, apoptosis, immune system development and immune response 16,17) by targeting messenger RNA (mRNA) during its translation into protein, either degrading the mRNA or inhibiting the translation process 18. Aberrant miRNA expression has been found to be associated with pathologies, including different types of cancer, inflammation and diabetes 19. Importantly, food-derived miRNA have been recently shown to be very stable in the gastrointestinal tract and be transferred to the blood circulation of adults, influencing gene expression in different tissues 20. In addition to tissues and cells, miRNAs have been isolated from body fluids, such as plasma, urine, saliva and tears 14. Further, exosomes, small cell-derived vesicles present in body fluids and carrying proteins and molecules, have been shown to take up miRNAs mediating their protection against digestion and facilitating their regulatory functions in different tissues and organs 21. Most recently, miRNAs have been isolated in high quantities from both animal and HM, and were shown to be present both as free molecules in skim milk 13,14,22–24 and packaged in vesicles such as milk exosomes and the
Journal of Ovarian Research
Background Among the various seasonal environmental changes, elevated ambient temperature during the summer season is a main cause of stress in dairy and beef cows, leading to impaired reproductive function and fertility. Follicular fluid extracellular vesicles (FF-EVs) play an important role in intrafollicular cellular communication by, in part, mediating the deleterious effects of heat stress (HS). Here we aimed to investigate the changes in FF-EV miRNA cargoes in beef cows in response to seasonal changes: summer (SUM) compared to the winter (WIN) season using high throughput sequencing of FF-EV-coupled miRNAs. In addition to their biological relevance, the potential mechanisms involved in the packaging and release of those miRNAs as a response to environmental HS were elucidated. Results Sequencing analysis revealed that an average of 6.6% of the EV-RNA mapped reads were annotated to bovine miRNAs. Interestingly, miR-148a, miR-99a-5p, miR-10b, and miR-143 were the top four miRNAs...
Differential expression of miRNAs in milk exosomes of cows subjected to group relocation
Research in Veterinary Science, 2018
This study examined microRNA (miRNAs) expression and regulatory patterns from milk exosomes of cows experienced group relocation, a common husbandry practice during the lactation period and used as a spontaneous model of stress. Total RNA from milk exosome samples was collected from 3 cows that showed an increased milk cortisol (HRC) after relocation (T2 vs T1) and 3 cows that did not show cortisol increase (LRC). A total of 69 known miRNAs were identified. Thirteen miRNAs were consistently down-regulated at T2 in comparison to T1, such as miR-2904-1, miR-142, miR-2284x and miR-30b-3p. Only two miRNAs, miR-2284z and miR-146a were significantly different between LRC and HRC group. Functional enrichment analyses highlighted that glucocorticoid receptor signaling and neurotrophic factor mediated TRK receptor signaling were among the biological pathways affected by differentially expressed miRNA target genes. Mir-135a-5p and miR-320a were involved in both biological pathways. miR-142-5p, miR-142-3p, miR-30b-5p and miR-320a shared the same target genes belonging to the RAS superfamily of small GTP-binding proteins (RAC1, RAP1A and RASA1), involved in neurotrophin-mediated cell survival. MiR-142, miR-135 and miR-320a in milk exosomes were the most responsive to group relocation of cows.