115 Exosome-mediated microRNA expression profile in follicular fluid of metabolically divergent postpartum cows

2019 ◽  
Vol 31 (1) ◽  
pp. 183
Author(s):  
T. Hailay ◽  
M. Hoelker ◽  
S. Gebremedhn ◽  
F. Rings ◽  
M. M. Saeed-Zidane ◽  
...  
Keyword(s):  
Energy Balance ◽  
Follicular Fluid ◽  
Mirna Expression ◽  
Cell Communication ◽  
Negative Energy ◽  
Negative Energy Balance ◽  
Differentially Expressed ◽  
Metabolic Status ◽  
Lactation Period ◽  
Down Regulation

Most high-milking cows enter a state of negative energy balance during the early lactation period. This phenomenon disturbs the metabolic status of the follicular fluid microenvironment, resulting in delayed ovulation. Cell-to-cell communication between the oocyte and the surrounding cells is crucial during folliculogenesis. Exosomes, evolutionarily conserved cargo molecules (30-150nm in diameter) carrying RNA and proteins, are known to be involved in cell-to-cell communication. Here, we aimed to investigate the association between postpartum metabolic status and the expression of exosomal microRNA (miRNA) in follicular fluid of Holstein-Friesian cows. For this, follicular fluid was collected from antral follicles (>8mm in diameter) using ovum pickup procedure from cows (n=30) on a weekly basis between weeks 5 and 10 postpartum. Follicular fluid collected from heifers (n=8) was used as a control. The energy status of each cow was assessed based on the blood metabolite (nonesterified fatty acids and β-hydroxybutyrate) concentration, body weight curve, and overall energy balance determined by dry matter intake. Afterwards, cows were categorized as early negative and late positive (cows show negative energy balance at early weeks and recovered at late weeks postpartum), always negative (cows did not recover until 15 weeks postpartum), and always positive (cows did not enter in to a state of negative energy balance). Following this, exosomes were isolated from pooled samples from each animal category using ultracentrifugation, and their morphology and size was characterised using electron microscopy and nanosight, respectively. Exosomal total RNA enriched with miRNA was isolated using an exosomal RNA isolation kit. Next-generation sequencing of miRNA was performed using Illumina NextSEqn 500 (Illumina Inc., San Diego, CA, USA). MicroRNAs with a fold change ≥2, P-value <0.05, and a false discovery rate of <0.1 were considered differentially expressed. The results showed that a total of 356 known and 156 novel miRNA were identified across samples. Differential expression analysis of miRNA between always-negative cows versus always-positive cows revealed down-regulation of all 6 differentially expressed miRNA, including bta-miR-451, bta-miR-132, and bta-miR-2285. Similarly, down-regulation of 14 miRNA, including bta-miR-20b, bta-miR-363, bta-miR-132, and bta-miR-451, and up-regulation of 3 miRNA was observed in always-negative cows compared to heifers. Furthermore, the target prediction analysis of the down-regulated miRNA have been shown to be involved in regulating different pathways including transforming growth factor-β signalling, cell cycle, hippo signalling, forkhead box O signalling, and endometrial cancer, among others. In conclusion, the results revealed that although negative energy balance in postpartum dairy cows suppressed exosomal miRNA expression in follicular fluid, the opposite was observed in metabolically unstressed cows. This divergence of exosome-mediated miRNA expression in the follicular fluid of metabolically stressed cows could be associated with the reduced fertility of those cows.

scholarly journals Alterations in hepatic miRNA expression during negative energy balance in postpartum dairy cattle

BMC Genomics ◽  
2014 ◽  
Vol 15 (1) ◽  
pp. 28 ◽  
Author(s):  
Attia Fatima ◽  
Sinead Waters ◽  
Padraig O’Boyle ◽  
Cathal Seoighe ◽  
Dermot G Morris
Keyword(s):  
Energy Balance ◽  
Dairy Cattle ◽  
Mirna Expression ◽  
Negative Energy ◽  
Negative Energy Balance

207 CIRCULATORY microRNA SIGNATURES IN FOLLICULAR FLUID IN RELATION TO THE GROWTH STATUS OF BOVINE OOCYTES

2013 ◽  
Vol 25 (1) ◽  
pp. 251 ◽  
Author(s):  
M. M. H. Sohel ◽  
D. Salilew-Wondim ◽  
M. Hölker ◽  
F. Rings ◽  
K. Schellander ◽  
...  
Keyword(s):  
Ingenuity Pathway Analysis ◽  
Pathway Analysis ◽  
Follicular Fluid ◽  
Mirna Expression ◽  
Cell Communication ◽  
Bioactive Molecules ◽  
Differentially Expressed ◽  
Oocyte Growth ◽  
Mirna Expression Pattern ◽  
Protein Ubiquitination

Cell-to-cell communication within the follicle involves many signalling molecules, and this process is believed to be mediated by secretion and uptake of exosomes that contained several bioactive molecules including circulatory microRNAs (miRNA). The present study was conducted to investigate the circulating miRNA expression pattern in exosome and nonexosomal portion of follicular fluid (FF) in follicles with fully grown or growing oocytes. For this, the FF and cumulus–oocyte complexes (COC) were retrieved from 5- to 8-mm individual follicles from ovaries obtained from local abattoir. Then, the oocytes were subjected to brilliant cresyl blue (BCB) staining and classified as BCB+ (fully grown oocytes) and BCB– (growing oocytes). Accordingly, the corresponding FF was classified as BCB+ and BCB– based on their oocyte source. Following this, the exosomes were trapped from each FF categories using ExoQuick™ (SBI System Bioscience). Thus, total RNA was isolated from exosomal and nonexosomal portion of the FF using miRNeasy mini kit (Qiagen) and subjected to miRNA expression studies. The human miRCURY LNA™ Universal RT miRNA PCR array system (Exiqon) was used for miRNA expression profiling. Data analysis was performed using a comparative threshold cycle (ΔCT) method. The results revealed that 26 miRNAs were found to be differentially expressed (fold change ≥2 and P < 0.05) between the exosomal portion of FF from fully grown and growing oocyte groups. Among these, 17 miRNA including miR-608, miR-654-5p, miR-640, miR-582-5p, miR-449b, miR-155 were upregulated and 9 miRNA including miR-373, miR-526b*, miR-33a*, miR-30b, miR-29a* were downregulated in exosomal portion of FF of growing oocyte groups. The ingenuity pathway analysis of genes predicted to be targeted by those miRNAs were found to be involved in WNT/β catenine, purine metabolism, protein ubiquitination, and cAMP-mediated signalling pathways. Similarly, 36 miRNA were differentially expressed between the non-exosomal portion of FF of fully grown and growing oocytes. From those, 27 miRNA including let-7i*, miR-328, miR-223, miR-19b-1*, miR-423-5p, miR-29c, miR-659 were upregulated, whereas the expression level of 9 miRNA including miR-381, miR-18a*, miR-30e*, miR-934, and miR-302c was downregulated in the nonexosomal portion of FF of growing oocyte groups. In addition, the ingenuity pathway analysis indicated that the genes predicted to be targeted by these miRNA were found to be involved in NRF2-mediated oxidative stress response, tight junction signalling, and protein ubiquitination pathways. In conclusion, this study detected the presence of exosome or non-exosome-mediated circulation of miRNA in the bovine follicular fluid and oocyte growth-dependent variation of circulatory miRNA in the follicular environment.

scholarly journals Mammary gene expression profiles during an intramammary challenge reveal potential mechanisms linking negative energy balance with impaired immune response

2010 ◽  
Vol 41 (2) ◽  
pp. 161-170 ◽  
Author(s):  
Kasey M. Moyes ◽  
James K. Drackley ◽  
Dawn E. Morin ◽  
Sandra L. Rodriguez-Zas ◽  
Robin E. Everts ◽  
...  
Keyword(s):  
Gene Expression ◽  
Immune Response ◽  
Energy Balance ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Negative Energy ◽  
Negative Energy Balance ◽  
Differentially Expressed ◽  
Positive Energy ◽  
Ad Libitum

Our objective was to compare mammary tissue gene expression profiles during a Streptococcus uberis ( S. uberis) mastitis challenge between lactating cows subjected to dietary-induced negative energy balance (NEB; n = 5) and cows fed ad libitum to maintain positive energy balance (PEB; n = 5) to better understand the mechanisms associated with NEB and risk of mastitis during the transition period. The NEB cows were feed-restricted to 60% of calculated net energy for lactation requirements for 7 days, and cows assigned to PEB were fed the same diet for ad libitum intake. Five days after feed restriction, one rear mammary quarter of each cow was inoculated with 5,000 cfu of S. uberis (O140J). At 20 h postinoculation, S. uberis-infected mammary quarters from all cows were biopsied for RNA extraction. Negative energy balance resulted in 287 differentially expressed genes (DEG; false discovery rate ≤ 0.05), with 86 DEG upregulated and 201 DEG downregulated in NEB vs. PEB. Canonical pathways most affected by NEB were IL-8 signaling (10 genes), glucocorticoid receptor signaling ( 13 ), and NRF2-mediated oxidative stress response ( 10 ). Among the genes differentially expressed by NEB, cell growth and proliferation ( 48 ) and cellular development ( 36 ) were the most enriched functions. Regarding immune response, HLA-A was upregulated due to NEB, whereas the majority of genes involved in immune response were downregulated (e.g., AKT1, IRAK1, MAPK9, and TRAF6). This study provided new avenues for investigation into the mechanisms relating NEB and susceptibility to mastitis in lactating dairy cows.

scholarly journals Gain and loss of subcutaneous and abdominal fat depot mass from late pregnancy to 100 days in milk in German Holsteins

2019 ◽  
Vol 86 (3) ◽  
pp. 296-302 ◽  
Author(s):  
Lena Ruda ◽  
Claudia Raschka ◽  
Korinna Huber ◽  
Reka Tienken ◽  
Ulrich Meyer ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Energy Balance ◽  
Energy Density ◽  
Negative Energy ◽  
Negative Energy Balance ◽  
Lactation Period ◽  
Dry Period ◽  
Gain And Loss ◽  
The Impact ◽  
Daily Gain

AbstractThis research paper addresses the hypothesis that in times of negative energy balance around parturition in dairy cattle, lipids stored in adipocytes are mobilised in a more intensive manner out of the abdominal depots than out of the subcutaneous adipose tissues. Furthermore, the impact of niacin supplementation and energy density of the ration on adipose tissue mass gain and loss was assessed. Absolute masses of subcutaneous (SCAT), retroperitoneal (RPAT), omental (OMAT), mesenterial (MAT) and abdominal adipose tissue as a whole (AAT) were estimated by ultrasonography at −42, 3, 21 and 100 DIM. Absolute and relative daily gain during dry period (−42 to 3 DIM) and loss in fresh cow period (3 to 21 DIM) and early lactation period (22 to 100 DIM) were calculated. Feeding regime neither by niacin nor by energy density exerted any effect on adipose tissue masses. The AAT was always bigger than SCAT, but RPAT, OMAT and MAT did not differ amongst each other. All depot masses showed similar patterns with an increase during dry period and a decrease after calving. In fresh cow period AAT absolutely and relatively lost more mass than SCAT. This confirms that AAT is more intensively mobilised than SCAT during that time span. Further absolute daily gain during dry period was strongly negatively correlated with absolute daily loss during fresh cow period. This underlines the impact of individual body condition on adipose mobilisation in periparturient dairy cows. According to these results, it has to be taken into account that the largest amount of fat mobilised in the fresh cow period origins from AAT. This might impact the pattern of adipose derived metabolites and metabolic effectors interacting in physiological and deregulated adaptation to negative energy balance.

scholarly journals PSI-1 A systematic review and meta-analysis of GWAS and gene expression results of Holstein cattle under negative energy balance and ketosis

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 265-266
Author(s):  
Riani Ananda Nunes Soares ◽  
Giovana Vargas ◽  
Malane M Muniz ◽  
Maria Amelia Menck Soares ◽  
Angela Canovas ◽  
...  
Keyword(s):  
Gene Expression ◽  
Systematic Review ◽  
Energy Balance ◽  
Gene Network ◽  
Meta Analysis ◽  
Significant Snps ◽  
Negative Energy ◽  
Negative Energy Balance ◽  
Differentially Expressed ◽  
Metabolic Conditions

Abstract Development of ketosis in high-producing dairy cows contributes to animal health issues and highlights the need for better understanding the genetic basis of metabolic diseases. Thus, the aim of this study was to evaluate the pattern of differential gene expression in liver of cows under negative energy balance (NEB), subclinical, and clinical ketosis through a systematic review and meta-analysis of published gene expression and genome-wide association studies (GWAS) results. After screening of the 118 articles found in the systematic review, 20 articles were included in the analysis. For this, 430 significant SNPs identified by GWAS were investigated to see if they were located within genes reported in gene expression studies. A permutation approach was used to identify the biological pathways associated with the metabolic conditions studied. A gene network was created using the differentially expressed genes harboring significant SNPs and a QTL enrichment analysis was performed to identify potential positional candidate loci. This study revealed 14 genes that are differentially expressed in the liver of cows in different metabolic conditions, which harbor 24 significant polymorphisms in reported GWAS. Three significant metabolic pathways were associated with NEB, subclinical and clinical ketosis. In addition, two important genes, PPARA and ACACA, were identified as differentially expressed in the three metabolic conditions. Gene network analysis revealed co-expression interactions among 34 genes associated with functions involving fatty acid transport and metabolism. The genes FN1 and PTK2 were enriched for QTL previously associated with the trait “ketosis” on chromosome 2 and with the trait “milk iron content” on chromosome 14, respectively. These findings improve the understanding of negative energy balance and ketosis in dairy cows, which could enhance selection for cows less susceptible to ketosis and help with the development of potential biomarkers for early diagnosis and prevention of ketosis.

scholarly journals The effect of a negative energy balance status on β-carotene availability in serum and follicular fluid of nonlactating dairy cows

2016 ◽  
Vol 99 (7) ◽  
pp. 5808-5819 ◽  
Author(s):  
J. De Bie ◽  
A. Langbeen ◽  
A.A.J. Verlaet ◽  
F. Florizoone ◽  
I. Immig ◽  
...  
Keyword(s):  
Energy Balance ◽  
Dairy Cows ◽  
Follicular Fluid ◽  
Negative Energy ◽  
Negative Energy Balance ◽  
Β Carotene

scholarly journals Follicular fluid proteomic profiling of dairy cows with anestrus caused by negative energy balance

2021 ◽  
Vol 20 (1) ◽  
pp. 650-663
Author(s):  
Chang Zhao ◽  
Yunlong Bai ◽  
Shixin Fu ◽  
Ling Wu ◽  
Chuang Xu ◽  
...  
Keyword(s):  
Energy Balance ◽  
Dairy Cows ◽  
Follicular Fluid ◽  
Negative Energy ◽  
Negative Energy Balance ◽  
Proteomic Profiling

scholarly journals Energy Balance Indicators during the Transition Period and Early Lactation of Purebred Holstein and Simmental Cows and Their Crosses

Animals ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 309
Author(s):  
Deise Aline Knob ◽  
André Thaler Neto ◽  
Helen Schweizer ◽  
Anna C. Weigand ◽  
Roberto Kappes ◽  
...  
Keyword(s):  
Energy Balance ◽  
Milk Yield ◽  
Milk Composition ◽  
Negative Energy ◽  
The Other ◽  
Negative Energy Balance ◽  
Metabolic Energy ◽  
Genetic Groups ◽  
Significant Difference ◽  
Fat Thickness

Crossbreeding in dairy cattle has been used to improve functional traits, milk composition, and efficiency of Holstein herds. The objective of the study was to compare indicators of the metabolic energy balance, nonesterified fatty acids (NEFA), beta-hydroxybutyrate (BHBA), glucose, body condition score (BCS) back fat thickness (BFT), as well as milk yield and milk composition of Holstein and Simmental cows, and their crosses from the prepartum period until the 100th day of lactation at the Livestock Center of the Ludwig Maximilians University (Munich, Germany). In total, 164 cows formed five genetic groups according to their theoretic proportion of Holstein and Simmental genes as follows: Holstein (100% Holstein; n = 9), R1-Hol (51–99% Holstein; n = 30), first generation (F1) crossbreds (50% Holstein, 50% Simmental; n = 17), R1-Sim (1–49% Holstein; n = 81) and Simmental (100% Simmental; n = 27). The study took place between April 2018 and August 2019. BCS, BFT blood parameters, such as BHBA, glucose, and NEFA were recorded weekly. A mixed model analysis with fixed effects breed, week (relative to calving), the interaction of breed and week, parity, calving year, calving season, milking season, and the repeated measure effect of cow was used. BCS increased with the Simmental proportion. All genetic groups lost BCS and BFT after calving. Simmental cows showed lower NEFA values. BHBA and glucose did not differ among genetic groups, but they differed depending on the week relative to calving. Simmental and R1-Sim cows showed a smaller effect than the other genetic groups regarding changes in body weight, BCS, or back fat thickness after a period of a negative energy balance after calving. There was no significant difference for milk yield among genetic groups, although Simmental cows showed a lower milk yield after the third week after calving. Generally, Simmental and R1-Simmental cows seemed to deal better with a negative energy balance after calving than purebred Holstein and the other crossbred lines. Based on a positive heterosis effect of 10.06% for energy corrected milk (ECM), the F1, however, was the most efficient crossbred line.

scholarly journals Metabolic Adaptations to Dynamic Energy Requirements during Lactation and Pregnancy in Dairy Cows with Varying Proportions of Holstein and Simmental Breed

Proceedings ◽  
2020 ◽  
Vol 73 (1) ◽  
pp. 9
Author(s):  
Deise Aline Knob ◽  
André Thaler Neto ◽  
Helen Schweizer ◽  
Anna Weigand ◽  
Roberto Kappes ◽  
...  
Keyword(s):  
Energy Balance ◽  
Dairy Cows ◽  
Fixed Effects ◽  
Mixed Model ◽  
Negative Energy ◽  
Negative Energy Balance ◽  
Repeated Measure ◽  
Holstein Cows ◽  
Mixed Model Analysis ◽  
Genetic Groups

Depending on the breed or crossbreed line, cows have to cope with a more or less severe negative energy balance during the period of high milk yields in early lactation, which can be detected by beta-hydroxybutyrate (BHBA) and non-esterified fatty acids (NEFAs) in blood. Preventing cows from undergoing a severe negative energy balance by breeding and/or feeding measures is likely to be supported by the public and may help to improve the sustainability of milk production. The aim was to compare BHBA and NEFA concentrations in the blood of Holstein and Simmental cows and their crosses during the prepartum period until the end of lactation. In total, 164 cows formed five genetic groups according to their theoretic proportion of Holstein and Simmental genes as follows: Holstein (100% Holstein; n = 9), R1-Hol (51–99% Holstein; n = 30), F1 crossbreds (50% Holstein, 50% Simmental; n = 17), R1-Sim (1–49% Holstein; n = 81) and Simmental (100% Simmental; n = 27). NEFA and BHBA were evaluated once a week between April 2018 and August 2019. A mixed model analysis with fixed effects breed, week (relative to calving), the interaction of breed and week, parity, calving year, calving season, milking season, and the repeated measure effect on cows was used. Holstein cows had higher NEFAs (0.196 ± 0.013 mmol/L), and Simmental cows had the lowest NEFA concentrations (0.147 ± 0.008 mmol/L, p = 0.03). R1-Sim, F1 and R1-Hol cows had intermediate values (0.166 ± 0.005, 0.165 ± 0.010, 0.162 ± 0.008 mmol/L; respectively). The highest NEFA value was found in the first week after calving (0.49 ± 0.013 mmol/L). BHBA did not differ among genetic groups (p = 0.1007). There was, however, an interaction between the genetic group and week (p = 0.03). While Simmental, R1-Sim and F1 cows had the highest BHBA value, the second week after calving (0.92 ± 0.07 and 1.05 ± 0.04, and 1.10 ± 0.10 mmol/L, respectively), R1-Hol and Holstein cows showed the BHBA peak at the fourth week after calving (1.16 ± 0.07 and 1.36 ± 0.12 mmol/L, respectively). Unexpectedly, Holstein cows had a high BHBA peak again at week 34 after calving (1.68 ± 0.21 mmol/L). The genetic composition of the cows affects NEFA and BHBA. Simmental and R1-Sim cows mobilize fewer body reserves after calving. Therefore, dairy cows with higher degrees of Simmental origin might be more sustainable in comparison with Holstein genetics in the present study.

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