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For volume 1 alone: A gold-standard collection of readily reproducible techniques for the molecular and genetic analysis of germ cells in a variety of different reproductive systems. Volume 1: Sperm and Oocyte Analysis focuses on sperm cells, oocyte analysis, oocyte maturation, fertilization, and preparation techniques. Highlights include in vitro maturation and fertilization of human, porcine, and canine oocytes; the cryopreservation of sperm cells; establishment of an in vitro spermatogenesis system; visualization of sperm accessory structures; and motility assays of stallion spermatozoa. Each readily reproducible protocol is described in step-by-step detail and contains an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. For both volumes: A gold-standard collection of readily reproducible techniques for the molecular and genetic analysis of germ cells in a variety of different reproductive systems. The methods cover sperm and egg activation, motility, fertilization, nuclear development, nuclear cloning, the molecular characterization of specific events, and the imaging of cell structures. Volume 1: Sperm and Oocyte Analysis focuses on sperm cells, oocyte analysis, oocyte maturation, fertilization, and preparation techniques. Volume 2: Molecular Embryo Analysis, Live Imaging, Transgenesis, and Cloning contains methods for molecular egg analysis, live egg imaging, nuclear cloning, oocyte preservation, and nuclear transfer. Comprehensive and cutting-edge, Germ Cell Protocols offers both novice and established researchers a gold-standard collection of hard-to-find methods of high impact research, diverse procedures that are easy-to-follow, well-illustrated, and allow a cross-species transfer of knowledge from lower vertebrates to higher mammalian systems.

Ed. Schatten, Heide

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Table of Contents

1. Cryopreservation of Semen of the Salmonidae with Special Reference to Large-Scale Fertilization (pp. 001-012)
   Lahnsteiner, Franz
2. Fertilization and Sperm Chemotaxis in Ascidians (pp. 013-026)
   Yoshida, Manabu
3. Assays for Vertebrate Sperm Chemotaxis (pp. 027-048)
   Sugiyama, Hitoshi; Al-Anzi, Bader; McGaughey, Robert W.; Chandler, Douglas E.
4. Stallion Spermatozoa Viability: Comparison of Flow Cytometry with Other Methods (pp. 049-058)
   Merkies, Katrina; Buhr, Mary M.
5. Visualization of Sperm Accessory Structures in the Mammalian Spermatids, Spermatozoa, and Zygotes by Immunofluorescence, Confocal, and Immunoelectron Microscopy (pp. 059-078)
   Sutovsky, Peter
6. Localization of Zona Pellucida Receptors on Live Sperm by Fluorophore-Conjugated Solubilized Zona Pellucida Proteins (pp. 079-094)
   Burkin, Heather R.; Paula Alves-Vieira, Ana; Miller, David J.
7. Detection of Human Papillomavirus DNA in Sperm Using Polymerase Chain Reaction (pp. 095-104)
   Olatunbosun, Olufemi A.; Case, Allison M.; Deneer, Harry G.
8. Spermatogonial Stem Cells (pp. 105-120)
   Nayernia, Karim; Li, Manyu; Engel, Wolfgang
9. Molecular and Cellular Approaches for Establishment of an In Vitro Spermatogenesis System (pp. 121-134)
   Nayernia, Karim; Engel, Wolfgang
10. Acrosin Activity Assay for the Evaluation of Mammalian Sperm Acrosome Reaction (pp. 135-140)
    Lax, Yehudit; Rubinstein, Sara; Breitbart, Haim
11. Isolation of Sea Urchin Sperm Plasma Membranes (pp. 141-150)
    Mengerink, Kathryn J.; Vacquier, Victor D.
12. Detection of Centrosome Structure in Fertilized and Artificially Activated Sea Urchin Eggs Using Immunofluorescence Microscopy and Isolation of Centrosomes Followed by Structural Characterization with Field Emission Scanning Electron Microscopy (pp. 151-164)
    Schatten, Heide; Chakrabarti, Amitabha
13. Microscopic Techniques for Studying Sperm–Oocyte Interaction During Fertilization and Early Embryonic Development (pp. 165-200)
    Tengowski, Mark W.
14. Immunocytochemical Staining of the Metaphase II Spindle in Mammalian Oocytes (pp. 201-214)
    Mullen, Steven F.; Critser, John K.
15. Spindle Imaging in Living Mammalian Oocytes Using the Liquid-Crystal Polarized Light Microscope and Its Practical Use (pp. 215-226)
    Wang, Wei-Hua; Gill, Jimmy; Boutin, Cathy
16. In Vitro Maturation and Fertilization of Pig Oocytes (pp. 227-234)
    Fan, Heng-Yu; Sun, Qing-Yuan
17. In Vitro Maturation and In Vitro Fertilization of Human Oocytes (pp. 235-254)
    Wang, Wei-Hua; Gill, Jimmy
18. In Vitro Maturation and Fertilization of Canine Oocytes (pp. 255-272)
    Reynaud, Karine; Saint-Dizier, Marie; Chastant-Maillard, Sylvie
19. Detection and Measurement of Membrane-Bound Protein Tyrosine Kinases in the Zebrafish Egg (pp. 273-284)
    Wu, Wenjun; Kinsey, William H.
20. Single-Cell Method for Estimation of Protein Tyrosine Kinases in the Zebrafish Egg (pp. 285-292)
    MacGregor, Elinor; Kinsey, William H.
21. Activity of MAPK/p90rsk During Fertilization in Mice, Rats, and Pigs (pp. 293-304)
    Sun, Qing-Yuan; Fan, Heng-Yu

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