Abstract
The ability to mature oocytes in vitro provides a tool for creating embryos by parthenogenesis, fertilization, and cloning. Unfortunately the quality of oocytes matured in vitro falls behind that of in vivo matured oocytes. To address this difference, transcriptional profiling by deep sequencing was conducted on pig oocytes that were either matured in vitro or in vivo. Alignment of over 18 million reads identified 1,316 transcripts that were differentially represented. One pathway that was overrepresented in the oocytes matured in vitro was for Wingless-type MMTV integration site (WNT) signaling. In an attempt to inhibit the WNT pathway, Dickkopf-related protein 1 was added to the in vitro maturation medium. Addition of Dickkopf-related protein 1 improved the percentage of oocytes that matured to the metaphase II stage, increased the number of nuclei in the resulting blastocyst stage embryos, and reduced the amount of disheveled segment polarity protein 1 protein in oocytes. It is concluded that transcriptional profiling is a powerful method for detecting differences between in vitro and in vivo matured oocytes, and that the WNT signaling pathway is important for proper oocyte maturation.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Signal Transducing / genetics
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Adaptor Proteins, Signal Transducing / metabolism
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Animals
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Blastocyst / cytology
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Blastocyst / metabolism
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Cell Nucleus / metabolism
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Cytoplasm / metabolism
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Dishevelled Proteins
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Embryo Transfer
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Female
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Fertilization in Vitro
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Gene Expression Profiling
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Gene Expression Regulation, Developmental
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High-Throughput Nucleotide Sequencing
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Intercellular Signaling Peptides and Proteins / genetics*
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Intercellular Signaling Peptides and Proteins / metabolism
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Intercellular Signaling Peptides and Proteins / pharmacology
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Metaphase
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Oocytes / cytology
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Oocytes / growth & development
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Oocytes / metabolism*
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Oogenesis / genetics*
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Parthenogenesis / genetics*
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Phosphoproteins / genetics
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Phosphoproteins / metabolism
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Swine
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Transcriptome
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Wnt Proteins / genetics*
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Wnt Proteins / metabolism
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Wnt Signaling Pathway / genetics*
Substances
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Adaptor Proteins, Signal Transducing
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Dishevelled Proteins
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Intercellular Signaling Peptides and Proteins
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Phosphoproteins
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Wnt Proteins
Grants and funding
Funding from Food for the 21st Century at the University of Missouri, and by the University of Missouri grant 13321 from the Missouri Life Science Research Board via Dr. Jerry Taylor. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.