Mammalian oocytes occupy a critical nexus in reproduction as they not only contribute half the genetic makeup of the embryo but also provide virtually all of the cytoplasmic building blocks required for sustaining embryogenesis. The journey that transforms a primordial germ cell into a mature oocyte (or egg) capable of fertilisation and embryonic development is of unrivalled complexity. The end-product of this remarkable developmental process is the "mature" egg, now replete with macromolecules and organelles, and poised to support embryonic development if fertilisation should occur. Given the importance of the oocyte for embryonic development, it is of no surprise that the oocyte holds centre stage in fertility clinics and greatly influences the outcome of assisted reproductive treatments (ARTs).
Mammalian Oocyte Regulation: Methods and Protocols provides a highly diverse compendium of detailed methodologies – primarily focusing on the murine model but also including chapters on human oocytes – for enabling researchers to interrogate every aspect of mammalian oocyte development including recombination, meiotic maturation, oocyte substrate uptake, chromosomal segregation and fertilisation.
Written in the highly successful Methods in Molecular Biology series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Mammalian Oocyte Regulation: Methods and Protocols serves as an ideal guide to novice oocyte researchers, experts who wish to add another dimension to their established portfolio, as well as the wider research community.
1. Studying Recombination in Mouse Oocytes Xianfei Sun and Paula E. Cohen
2. Analysis of Recombinants in Female Mouse Meiosis Esther de Boer, Maria Jasin, and Scott Keeney
3. Studying Meiosis Specific Cohesins in Mouse Embryonic Oocytes Ji-hye Kim, Kei-ichiro Ishiguro, Nobuaki Kudo, and Yoshinori Watanabe
4. Immunohistochemical Approaches to the Study of Human Fetal Ovarian Development Jing He, Andrew J. Childs, Jieqian Zhou, and Richard A. Anderson
5. Protein Kinase Assays for Measuring MPF and MAPK Activities in Mouse and Rat Oocytes and Early Embryos Jacek Z. Kubiak
6. Time-Lapse Epi-Fluorescence Imaging of Expressed cRNA to Cyclin B1 for Studying Meiosis I in Mouse Oocytes Janet E. Holt, Simon I.R. Lane, and Keith T. Jones
7. Using FRET to Study RanGTP Gradients in Live Mouse Oocytes Julien Dumont and Marie-Helene Verlhac
8. Making cRNA for Microinjection and Expression of Fluorescently-Tagged Proteins for Live-Cell Imaging in Oocytes Mark Levasseur
9. RNAi-Based Methods for Gene Silencing in Mouse Oocytes Paula Stein, Petr Svoboda, and Richard M. Schultz
10. Micro-Injection of Morpholino Oligonucleotides for Depleting Securin in Mouse Oocytes Petros Marangos
11. Measuring Transport and Accumulation of Radiolabelled Substrates in Oocytes and Embryos Jay M. Baltz, Hannah E. Corbett, and Samantha Richard
12. Immunofluorescence Staining of Spindles, Chromosomes and Kinetochores in Human Oocytes Solon Riris, Suzanne Cawood, Paul Serhal, and Hayden Homer
13. Studying the Roles of Aurora-C Kinase During Meiosis in Mouse Oocytes Kuo-Tai Yang, Yi-Nan Lin, Shu-Kuei Li, and Tang K. Tang
14. Chromosome Spreads with Centromere Staining in Mouse Oocytes Jean-Philippe Chambon, Khaled Hached, and Katja Wassmann
15. Preparation of Mammalian Oocytes for Transmission Electron Microscopy Lynne Anguish and Scott Coonrod
16. Measuring Ca2+ Oscillations in Mammalian Eggs Karl Swann
17. Counting Chromosomes in Intact Eggs Teresa Chiang and Michael A. Lampson
18. Free-Hand Bisection of Mouse Oocytes and Embryos Zbigniew Polanski and Jacek Z. Kubiak
19. Microarray-CGH for the Assessment of Aneuploidy in Human Polar Bodies and Oocytes Souraya Jaroudi and Dagan Wells
20. Nuclear Transfer in the Mouse Oocyte Eiji Mizutani, Atsuo Ogura, and Teruhiko Wakayama Petros Marangos