144. ROLE OF RNA-BINDING PROTEIN, MUSASHI-1 (MSI-1), IN MURINE FOLLICULOGENESIS AND OOCYTE DEVELOPMENT
K. M. Gunter A , B. A. Fraser A B , A. P. Sobinoff A , N. A. Siddall B C , G. R. Hime B C and E. A. McLaughlin A BA School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW, Australia
B School of Environmental & Life Sciences, University of Newcastle, ARC Centre of Excellence for Biotechnology and Development, Callaghan, NSW, Australia
C The University of Melbourne, Department of Anatomy and Cell Biology, Parkville, VIC, Australia
Reproduction, Fertility and Development 21(9) 62-62 https://doi.org/10.1071/SRB09Abs144
Published: 26 August 2009
Abstract
Follicular development and oocyte maturation in mammals requires the temporal and spatial control of protein production. Consequently, it is hypothesised that the preovulatory follicle represses mRNA translation until specific proteins are required during oocyte maturation. Increasingly RNA-binding proteins are being recognised as important contributors to germ cell development, particularly during oocyte transcriptional quiescence. We have identified the presence of RNA-binding protein musashi-1 (Msi-1) mRNA within the mouse ovary and mature mouse oocyte, where the protein is believed to act as a translational repressor by binding to specific sequences within the 3' UTR of target mRNA molecules. Recent studies in various mammalian systems have identified p21 WAF1, cdkn2a, notch and m-numb as potential targets of Msi-1. We have also identified morf4l1 as a potential target through preliminary pulldown and microarray analysis using a GST tagged Msi-1 recombinant protein. To further study these potential targets, a transgenic Msi-1 mouse was produced to overexpress the RNA-binding protein in the developing oocyte. Real time PCR, performed on intact ovaries of WT and Tg mice, has so far demonstrated a 1.5-fold increase in Msi-1 expression in tgMsi-1/+ ovaries, above WT ovary expression. Real time PCR analysis of Msi-1 target mRNA expression has also shown an overall increase in expression in the tgMsi-1/+ ovaries of p21 WAF1 (~2.5-fold), cdkn2a (~2-fold), and notch (~3-fold). However m-numb and morf4l1 do not appear to be targets of Msi-1 in the oocyte, with no significant difference in expression between the WT and tgMsi-1/+ ovaries analysed. Functional quantification of oocyte development reveals a significantly less oocytes produced from superovulated juvenile mice compared with wild type litter mates. Therefore, preliminary analysis suggests that Msi-1 may play a role in binding the transcripts of genes necessary for cell cycle regulation and chromatin remodelling, characteristic of meiotic progression and oocyte development.
