Effective protein inhibition in intact mouse oocytes through peptide nanoparticle-mediated antibody transfection
- Published
- Accepted
- Subject Areas
- Biotechnology, Cell Biology, Women's Health
- Keywords
- meiosis, peptide nanoparticle, mouse oocytes, antibody transfection, protein inhibition
- Copyright
- © 2015 Li et al.
- Licence
- This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ PrePrints) and either DOI or URL of the article must be cited.
- Cite this article
- 2015. Effective protein inhibition in intact mouse oocytes through peptide nanoparticle-mediated antibody transfection. PeerJ PrePrints 3:e1566v1 https://doi.org/10.7287/peerj.preprints.1566v1
Abstract
Female meiosis is a fundamental area of study in reproductive medicine, and the mouse oocyte model of in vitro maturation (IVM) to study female meiosis is the most widely used. To investigate the probable role(s) of an unknown protein in female meiosis, the method traditionally used involves microinjecting a specific antibody into mouse oocytes. Recently, in studies on somatic cells, peptide nanoparticle-mediated antibody transfection has become a popular tool because of its high efficiency, low toxicity, good stability, and strong serum compatibility. However, till now, no researchers have tried using this technique on mouse oocytes because the zona pellucida surrounding the oocyte membrane (vitelline membrane) is usually thought or proved to be a tough barrier to macromolecules, such as antibodies and proteins. Therefore, we attempted to introduce an antibody into mouse oocytes using a peptide nanoparticle. Here we show for the first time that with our optimized method, an antibody can be effectively delivered into mouse oocytes and inhibit its target protein with high specificity. We obtained significant results using small GTPase Arl2 as a test subject protein. We propose peptide nanoparticle-mediated antibody transfection to be a superior alternative to antibody microinjection for preliminary functional studies of unknown proteins in mouse oocytes.
Author Comment
This manuscript has been submitted to Peer J as a research article for peer review. We believe the method described in this manuscript will benefit many researchers in mammalian female meiosis. In this hope, any response or communication, which may help further improve this method, is welcome.
Supplemental Information
Raw data for Figure 1B
This file includes three sheets: "Original data" sheet has rhodamine fluorescence intensity measurements for all examined oocytes. Net rhodamine fluorescence intensity is equal to cytoplasmic intensity subtracted by background intensity (outside the oocyte); "Statistics" sheet has all the data for statistics and the P value from T test. "Graph" sheet has AVEs and SEMs for the graph corresponding to Figure 1B.
Raw data corresponding to Figure 3B
This file includes three sheets: "Original data" sheet has numbers and percentages of oocytes at each stage (GV, GV-like, Pre-MI, MI, AI and TI) at 8 hour of IVM from three repeats; "Statistics" sheet has all the GV-like or MI percentages for statistics and the P value from T test; "Graph" sheet has the AVEs and SEMs for the graph corresponding to Figure 3B.
Raw data corresponding to Figure 3D
This file includes three sheets: "Original data" sheet has all the measurements of MI spindle length or width (in pixel) at 8 hour of IVM from three repeats; "Statistics" sheet has all the measurements of MI spindle length (in pixel) for statistics and the P value from T test. Statistics for spindle width is not shown since it's not significant; "Graph" sheet has all the AVEs and SEMs (in pixel or μm) for the graph corresponding to Figure 3D.
Raw data corresponding to Figure 3F
This file includes three sheets: "Original data" sheet has all the numbers and percentages of oocytes at each stage (GV, GV-like, Pre-MI, MI, AI and TI) at 8 hour of IVM from three repeats; "Statistics" sheet has all the Pre-MII or total MII (MII) percentages for statistics and the P value from T test; "Graph" sheet has all the AVEs and SEMs for the graph corresponding to Figure 3F.