The role of microRNAs in regulation of bovine ovarian follicular development
Final Report Abstract
In this the first phase of the this project, we have provided detailed insights about the miRNA enrichment and degradation in the granulosa cells of subordinate and dominant follicle during the early luteal phase and at later follicular phase of bovine estrous cycle using high-throughput miRNA sequencing technology. Thus, it was revealed that including bta-miR-449a, bta-miR-449c and bta-miR-222 a total of 16 miRNAs were differentially between the granulosa cells of subordinate and dominant follicle at day 3 of the estrous cycle. However, the number of differentially expressed miRNAs between the two granulosa was increased to 108 at day 7 of the estrous cycle, indicating the presence of distinct miRNA expression profile differences between the granulosa cells of dominant and subordinate follicles at day 7 compared to day 3 of the estrous cycle. In addition, compared to the subordinate follicles, the granulosa cells of dominant follicles displayed a marked temporal miRNA expression patterns during the early luteal phase of the bovine estrous cycle. Functional annotation using a bioinformatics approaches indicated that the differentially expressed miRNAs were found to be involved in major signaling which are believed to be associated with follicular development, namely, Wnt signaling, TGF-beta signaling, oocyte meiosis and GnRH signaling A similar analysis at day 19 of the estrous cycle (at late stage of follicular phase) including miR-183~96~182, miR-132 and miR-17- 92 miRNA clusters, the expression levels of 64 miRNAs were found to significantly different between the granulosa cells of preovulatory dominant and subordinate follicles. Thus, in the first phase of the experiment, we have identified several miRNAs which could be associated with follicular recruitment, selection and dominance in the early luteal phase and follicle dominance and ovulation in later follicular phase of the bovine estrous cycle. Therefore, to validate the function of those candidate miRNAs, we have selected miR- 183~96~182 and miRNAs miR-17-92 cluster miRNAs and their functional role was investigated in bovine granulosa cells cultured in vitro using gain and loss function. Accordingly, the in vitro functional study has revealed that the miR-183~96~182 cluster miRNAs were found involve in bovine granulosa cell proliferation and cell cycle transition by coordinately targeting Foxo1 gene expression while microRNA 17-92 cluster miRNAs were involved in bovine granulosa cells proliferation, differentiation and progesterone production by targeting PTEN and BMPR2 genes. Therefore, with specific project, we have generated substantial and valuable data that could give an insight into the expression patterns and the role of miRNAs in regulation of bovine ovarian follicular development.
Publications
- (2014): Expression profiling of noncoding microRNAs in bovine granulosa cells of preovulatory dominant follicle using deep sequencing. 40th Annual Conference of the International Embryo Transfer Society (IETS), 11.- 14.1.2014, Reno NV, USA, Reprod Fert Dev 26, 1: 113
Gebremedhn S, Ahmad I, Salilew-Wondim D, Sahadevan S, Hoelker M, Rings F, Uddin M J, Tholen E, Looft C, Schellander K, Tesfaye D
- (2014): Investigation of miRNAs enrichment and degradation in bovine granulosa cells during follicular development (Untersuchung der miRNA Anreicherung und Abbau in bovine Granulosazellen während der Follikelreifung)
Ahmad, Ijaz
- (2014): Regulatory microRNA enrichment and degradation in granulosa cells during bovine follicular recruitment and dominance. 40th Annual Conference of the International Embryo Transfer Society (IETS), 11.-14.1.2014, Reno NV, USA, Reprod Fert Dev 26, 1: 111
Salilew-Wondim D, Ahmad I, Gebremedhn S, Sahadevan S, Hoelker M, Rings F, Uddin M J, Tholen E, Looft C, Schellander K, Tesfaye D
- 2014. The expression pattern of microRNAs in granulosa cells of subordinate and dominant follicles during the early luteal phase of the bovine estrous cycle. PloS one 9(9):e106795
Salilew-Wondim D, Ahmad I, Gebremedhn S, Sahadevan S, Hossain MD, Rings F, Hoelker M, Tholen E, Neuhoff C, Looft C, Schellander K, Tesfaye D
(See online at https://doi.org/10.1371/journal.pone.0106795) - 2015. MicroRNA Expression Profile in Bovine Granulosa Cells of Preovulatory Dominant and Subordinate Follicles during the Late Follicular Phase of the Estrous Cycle. PloS one 10(5):e0125912
Gebremedhn S, Salilew-Wondim D, Ahmad I, Sahadevan S, Hossain MM, Hoelker M, Rings F, Neuhoff C, Tholen E, Looft C, Schellander K, Tesfaye D
(See online at https://doi.org/10.1371/journal.pone.0125912) - 2016. MicroRNA 17–92 cluster regulates proliferation and differentiation of bovine granulosa cells by targeting PTEN and BMPR2 genes. Cell & Tissue Research
Andreas E, Hoelker M, Neuhoff C, Tholen E, Schellander K, Tesfaye D, Salilew-Wondim D
(See online at https://doi.org/10.1007/s00441-016-2425-7) - 2016. MicroRNA-183~96~182 Cluster Regulate Bovine Granulosa Cell Proliferation and Cell Cycle Transition by Coordinately Targeting FOXO1. Biol Reprod April 27, 115.137539
Gebremedhn S, Salilew-Wondim D, Hoelker M, Rings F, Neuhoff C, Tholen E, Schellander K, Tesfaye D
(See online at https://doi.org/10.1095/biolreprod.115.137539)
