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Drosha Expression and Microprocessor Activity in Maturing Bovine Oocytes and Cultured Granulosa Cells

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Title: Drosha Expression and Microprocessor Activity in Maturing Bovine Oocytes and Cultured Granulosa Cells
Author: Stuart, Deirdre
Department: Department of Biomedical Sciences
Program: Biomedical Sciences
Advisor: LaMarre, Jonathan
Abstract: This thesis investigates localization and expression of RNA-binding protein Drosha within maturing bovine oocytes, and the effect of experimentally inhibiting Drosha on processing several microRNA precursors into mature forms in cumulus cells (CCs). MicroRNAs regulate gene expression post-transcriptionally by binding messenger RNAs (mRNAs), causing translational suppression and degradation. MicroRNA synthesis is mediated by the Microprocessor complex containing Drosha, which cleaves primary transcripts to precursor microRNAs. Previous work demonstrated a marked increase in Drosha mRNA and protein within bovine oocytes over maturation and zygotic development. To further characterize the role of Drosha, expression and localization during maturation were assessed using immunofluorescence microscopy, qRT-PCR, and meiotic inhibition. To address functional reproductive roles of Drosha, microRNA profiles in Drosha-siRNA-silenced CC culture were evaluated. Together, these studies suggest Drosha expression in the oocyte-surrounding environment is regulated at the gene expression level and by physical localization within oocytes, and participates in microRNA biogenesis in CCs.
URI: https://hdl.handle.net/10214/26564
Date: 2021-12
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Embargoed Until: 2022-12-07


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