MicroRNAs from Extracellular Vesicles Secreted by Bovine Embryos as Early Biomarkers of Developmental Competence
Abstract
:1. Introduction
2. Results
2.1. Identification and Characterization of EVs from Conditioned-Culture Media by Early Embryos
2.2. miRNAs Differential Expression Analysis
3. Discussion
4. Materials and Methods
4.1. General Design
4.2. In Vitro Embryo Production
4.3. Separation of EVs from Conditioned-Culture Media by Early Embryos
4.4. Morphological and Molecular Characterization of Nanoparticles from Conditioned-Culture Media by Early Embryos
4.4.1. Flow Cytometer for EVs Markers
4.4.2. Transmission Electron Microscopy (TEM) Analysis
4.4.3. Nanoparticles Tracking Analysis (NTA)
4.5. miRNAs Cargo Characterization of EVs from Conditioned-Culture Media by Early Embryos
4.5.1. Extracellular Vesicles RNA Isolation and Quantification
4.5.2. Small RNAs Sequencing Analysis
4.6. Statistical Analysis
Author Contributions
Funding
Conflicts of Interest
Abbreviations
EVs | Extracellular vesicles |
miR | microRNA |
TEM | Transmission electron microscopy |
NTA | Nanoparticles tracking analysis |
NGS | Next-generation sequencing |
G1 | Group 1: embryos arrested on 8–16-cells stage at day 3.5 of culture |
G2 | Group 2: embryos that reached blastocyst stage at day 7 of culture |
IVF | In vitro fertilization |
EGA | Embryo genome activation |
SOF | Synthetic oviduct fluid |
SOFd | EVs-depleted SOF |
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miR | Significance Comparing G1 vs. G2 | Gene Expression Respect to G2 |
---|---|---|
bta-miR-103 | 0.0003 | Increased |
bta-miR-100 | 0.0004 | Increased |
bta-miR-502a | 0.0097 | Increased |
bta-miR-1 | 0.0208 | Increased |
bta-miR-101 | 0.1199 | Increased |
bta-let-7e | 0.1682 | Increased |
bta-miR-215 | 0.1876 | Increased |
bta-miR-205 | 0.2915 | Increased |
bta-miR-193a-3p | 0.4369 | Increased |
bta-miR-224 | 0.4895 | Increased |
bta-miR-222 | 0.0034 | Decreased |
bta-miR-92a | 0.0323 | Decreased |
bta-miR-2285av | 0.0385 | Decreased |
bta-miR-140 | 0.0429 | Decreased |
bta-miR-25 | 0.0545 | Decreased |
bta-miR-9-5p | 0.0692 | Decreased |
bta-miR-16b | 0.0824 | Decreased |
bta-miR-191 | 0.1003 | Decreased |
bta-miR-744 | 0.1038 | Decreased |
bta-miR-451 | 0.1163 | Decreased |
bta-miR-10b | 0.1211 | Decreased |
bta-let-7f | 0.1519 | Decreased |
bta-miR-2904 | 0.1561 | Decreased |
bta-miR-11980 | 0.1609 | Decreased |
bta-let-7g | 0.1609 | Decreased |
bta-miR-30d | 0.1714 | Decreased |
bta-miR-93 | 0.1873 | Decreased |
bta-miR-378 | 0.1994 | Decreased |
bta-let-7i | 0.2239 | Decreased |
bta-miR-148b | 0.2253 | Decreased |
bta-miR-143 | 0.2362 | Decreased |
bta-miR-144 | 0.2412 | Decreased |
bta-miR-151-3p | 0.2510 | Decreased |
bta-miR-21-5p | 0.2891 | Decreased |
bta-miR-221 | 0.3007 | Decreased |
bta-miR-152 | 0.3234 | Decreased |
bta-miR-455-5p | 0.3246 | Decreased |
bta-miR-199a-3p | 0.3564 | Decreased |
bta-miR-11995 | 0.3646 | Decreased |
bta-miR-200a | 0.3960 | Decreased |
bta-miR-96 | 0.4180 | Decreased |
bta-miR-124b | 0.4465 | Decreased |
bta-miR-124a | 0.4622 | Decreased |
bta-miR-15b | 0.4951 | Decreased |
bta-miR-92b | 0.5279 | Decreased |
bta-miR-486 | 0.1387 | Equal |
bta-miR-10a | 0.2450 | Equal |
bta-miR-423-5p | 0.3259 | Equal |
bta-let-7c | 0.3432 | Equal |
bta-miR-27a-3p | 0.3529 | Equal |
bta-miR-1246 | 0.3565 | Equal |
bta-miR-27b | 0.3948 | Equal |
bta-let-7b | 0.4056 | Equal |
bta-miR-183 | 0.4368 | Equal |
bta-miR-151-5p | 0.5007 | Equal |
bta-miR-142-5p | 0.5462 | Equal |
bta-miR-302b | 0.5876 | Equal |
bta-miR-24-3p | 0.5886 | Equal |
bta-miR-181a | 0.5970 | Equal |
bta-miR-29b | 0.5970 | Equal |
bta-miR-99a-5p | 0.6146 | Equal |
bta-miR-29d-3p | 0.6151 | Equal |
bta-miR-29c | 0.6178 | Equal |
bta-miR-320a | 0.6476 | Equal |
bta-miR-125b | 0.6481 | Equal |
bta-miR-192 | 0.6536 | Equal |
bta-miR-29a | 0.6548 | Equal |
bta-miR-125a | 0.6586 | Equal |
bta-miR-484 | 0.6634 | Equal |
bta-miR-26b | 0.6787 | Equal |
bta-miR-199a-5p | 0.6898 | Equal |
bta-miR-30a-5p | 0.7072 | Equal |
bta-miR-22-3p | 0.7340 | Equal |
bta-miR-146a | 0.7652 | Equal |
bta-miR-375 | 0.7673 | Equal |
bta-miR-2478 | 0.7729 | Equal |
bta-miR-122 | 0.7755 | Equal |
bta-miR-141 | 0.7853 | Equal |
bta-miR-30e-5p | 0.7925 | Equal |
bta-miR-146b | 0.7971 | Equal |
bta-miR-26a | 0.8062 | Equal |
bta-miR-148a | 0.8207 | Equal |
bta-miR-99b | 0.8406 | Equal |
bta-miR-184 | 0.8484 | Equal |
bta-miR-7 | 0.8697 | Equal |
bta-miR-23b-3p | 0.8705 | Equal |
bta-miR-423-3p | 0.8750 | Equal |
bta-miR-30c | 0.8867 | Equal |
bta-miR-10174-3p | 0.8955 | Equal |
bta-let-7a-5p | 0.9635 | Equal |
bta-miR-182 | 0.9746 | Equal |
bta-miR-23a | 0.9817 | Equal |
bta-miR-186 | 0.9912 | Equal |
bta-miR-185 | 1 | Equal |
bta-miR-2473 | 1 | Equal |
KEEG Pathway | p-Value | Target Genes | miRNA |
---|---|---|---|
Fatty acid biosynthesis | 0.000005 | 1 | 1 |
Fatty acid metabolism | 0.00017 | 4 | 2 |
Lysine degradation | 0.00089 | 9 | 4 |
Gap junction | 0.00089 | 19 | 4 |
Signaling pathways regulating pluripotency of stem cells | 0.0046 | 27 | 4 |
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Melo-Baez, B.; Wong, Y.S.; Aguilera, C.J.; Cabezas, J.; Mançanares, A.C.F.; Riadi, G.; Castro, F.O.; Rodriguez-Alvarez, L. MicroRNAs from Extracellular Vesicles Secreted by Bovine Embryos as Early Biomarkers of Developmental Competence. Int. J. Mol. Sci. 2020, 21, 8888. https://doi.org/10.3390/ijms21238888
Melo-Baez B, Wong YS, Aguilera CJ, Cabezas J, Mançanares ACF, Riadi G, Castro FO, Rodriguez-Alvarez L. MicroRNAs from Extracellular Vesicles Secreted by Bovine Embryos as Early Biomarkers of Developmental Competence. International Journal of Molecular Sciences. 2020; 21(23):8888. https://doi.org/10.3390/ijms21238888
Chicago/Turabian StyleMelo-Baez, Bárbara, Yat S. Wong, Constanza J. Aguilera, Joel Cabezas, Ana C. F. Mançanares, Gonzalo Riadi, Fidel O. Castro, and Lleretny Rodriguez-Alvarez. 2020. "MicroRNAs from Extracellular Vesicles Secreted by Bovine Embryos as Early Biomarkers of Developmental Competence" International Journal of Molecular Sciences 21, no. 23: 8888. https://doi.org/10.3390/ijms21238888