MicroRNA-34 family expression in bovine gametes and preimplantation embryos
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RESEARCH
Open Access
MicroRNA-34 family expression in bovine gametes and preimplantation embryos Allison Tscherner1, Graham Gilchrist1, Natasha Smith1, Patrick Blondin2, Daniel Gillis3 and Jonathan LaMarre1*
Abstract Background: Oocyte fertilization and successful embryo implantation are key events marking the onset of pregnancy. In sexually reproducing organisms, embryogenesis begins with the fusion of two haploid gametes, each of which has undergone progressive stages of maturation. In the final stages of oocyte maturation, minimal transcriptional activity is present and regulation of gene expression occurs primarily at the post-transcriptional level. MicroRNAs (miRNA) are potent effectors of post-transcriptional gene silencing and recent evidence demonstrates that the miR-34 family of miRNA are involved in both spermatogenesis and early events of embryogenesis. Methods: The profile of miR-34 miRNAs has not been characterized in gametes or embryos of Bos taurus. We therefore used quantitative reverse transcription PCR (qRT-PCR) to examine this family of miRNAs: miR-34a, -34b and -34c as well as their precursors in bovine gametes and in vitro produced embryos. Oocytes were aspirated from antral follicles of bovine ovaries, and sperm cells were isolated from semen samples of 10 bulls with unknown fertility status. Immature and in vitro matured oocytes, as well as cleaved embryos, were collected in pools. Gametes, embryos and ovarian and testis tissues were purified for RNA. Results: All members of the miR-34 family are present in bovine spermatozoa, while only miR-34a and -34c are present in oocytes and cleaved (2-cell) embryos. Mir-34c demonstrates variation among different bulls and is consistently expressed throughout oocyte maturation and in the embryo. The primary transcript of the miR-34b/c bicistron is abundant in the testes and present in ovarian tissue but undetectable in oocytes and in mature spermatozoa. Conclusions: The combination of these findings suggest that miR-34 miRNAs may be required in developing bovine gametes of both sexes, as well as in embryos, and that primary miR-34b/c processing takes place before the completion of gametogenesis. Individual variation in sperm miR-34 family abundance may offer potential as a biomarker of male bovine fertility. Keywords: Gametogenesis, MicroRNA, Biomarkers
Background Terminal maturation is the final series of nuclear and cytoplasmic events that bring a haploid gamete to a state where it has the capability to contribute to a fertilization event and establish an embryo. The maturing oocyte undergoes dynamic morphological and nuclear rearrangements as it approaches ovulation, progressively decreasing its transcriptional activities [1] together with the condensation of genetic material. At this time, the oocyte contains a repertoire of RNAs that will guide it * Correspondence: [email protected] 1 Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, 50 Stone Road East, Guelph, Canada Full list of author information is avai
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