| 1:Identification of miRNAs during mouse postnatal ovarian development and superovulation.Small Non coding RNAs known as MicroRNAs play vital roles in regulation of gene expression in broad range of tissues, that also the ovary at post-transcriptional level through sequence complementarity. Strict regulation of lot number of genes involved in ovarian development and folliculogenesis might be regulated by these miRNAs at transcription level. Therefore, identification of miRNAs in specific tissues is considered a very important step towards understanding the role of miRNAs in different biological processes. To investigate the role of microRNAs during ovarian development and folliculogenesis, we sequenced eight different libraries using Illumina deep sequencing technology at different stages of gonadal maturation with/without treatment of PMSG/hCG for superovulation.From massive sequencing reads, clean reads of 16-26 bp were selected for further analysis of differential expression analysis and novel microRNA annotation. Expression analysis of all miRNAs at different stages of ovarian developmental showed that some miRNAs were differentially expressed at specific developmental stage while others were present ubiquitously. Out of all differentially expressed miRNAs we reported 61 miRNAs with a fold change of more than 2 at different developmental stages among all libraries. Among differentially expressed miRNAs we reported 61 miRNAs with a fold change of more than 2 at different developmental stages among all libraries. Among the up-regulated miRNAs, mmu-miR-1298 had the highest fold change with 4.025 while mmu-miR-150 was down-regulated more than 3 fold. Furthermore, we found 2659 target genes for 20 differentially expressed microRNAs using seven different target predictions programs (DIANA-mT, miRanda, miRDB, miRWalk, RNAhybrid, PICTAR5, TargetScan). Analysis of the predicted targets showed certain ovary specific genes targeted by single or multiple microRNAs. Furthermore, pathway annotation and Gene ontology showed involvement of these microRNAs in basic cellular process.These results show the presence of different miRNAs at different stages of postnatal ovarian development and superovulation. Potential role of these microRNAs was elucidated using bioinformatics tools in regulation of different pathways, biological functions and cellular components underlying ovarian development and superovulation. Our results provide a baseline for further analysis of miRNAs and their specific roles during ovarian development and superovulation. Furthermore, present study provides a base for characterization and functional validation of individual miRNAs to discover their role in ovarian development and female fertility.2:microRNA-484 mediated post transcriptional gene regulation in mouse granulosa cells.During the process of ovarian growth and follicular development, many follicles go through follicular atresia and eventually only few follicles accomplish ovulation. Studies shows that follicular atresia is mainly due to granulosa cells apoptosis, but mechanism of granulosa cells apoptosis at molecular is still not clear. Therefore, to study the molecular mechanism especially modifications at post-transcriptional levels, we first elaborated the expression profile miR-484 in granulosa cells at different stages through qRT-PCR. Bases on the expression pattern we further used miR-484 mimics and inhibitors to see the changes in the expression level of miR484. Our results show that over expression or decreased in expression of miR-484 influence the expression of its target gene Acvrlb thus shows its involvement in granulosa cells growth. |
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