Germ Cell Protocols Volume 1: Sperm and Oocyte Analysis

All sexually reproducing organisms produce primordial germ cells, a small population of cells that differentiate into gametes of either sex and carry to- potency, an ability to develop into an entire new organism. The study of germ cells has undergone enormous advances in recent years and has entered into an explosive phase of new discoveries with the introduction of transgenic te- nologies and nuclear cloning. Basic knowledge and techniques developed for lower vertebrate and invertebrate systems have facilitated the study of higher vertebrates, including humans. Many experiments that have first been performed on lower vertebrates provided the tools and strategies that could later be applied to other less readily available mammalian systems. The discovery of centrosomes in ascidians and sea urchin eggs now benefits studies of fertility and infertility in mammals. External in vitro fertilization, now a common technique in assisted fertilization, has only been possible as a result of numerous studies in lower systems in which external fertilization is natural. Egg activation, first explored in sea urchin and ascidian eggs, now benefits techniques designed to increase cl- ing efficiency in farm and domestic animals. Gene manipulations and molecular methods have added to the possibilities of producing live offspring with en- mous biomedical, ecological, and economic implications. The two volumes on germ cells combine techniques in a variety of d- ferent systems; we have selected those systems that have provided landmarks in advancing our knowledge on germ cells.

Cryopreservation of Semen of the Salmonidae with Special Reference to Large-Scale Fertilization Franz Lahnsteiner Fertilization and Sperm Chemotaxis in Ascidians Manabu Yoshida Assays for Vertebrate Sperm Chemotaxis Hitoshi Sugiyama, Bader Al-Anzi, Robert W. McGaughey, and Douglas E. Chandler Stallion Spermatozoa Viability: Comparison of Flow Cytometry with Other Methods Katrina Merkies and Mary M. Buhr Visualization of Sperm Accessory Structures in the Mammalian Spermatids, Spermatozoa, and Zygotes by Immunofluorescence, Confocal, and Immunoelectron Microscopy Peter Sutovsky Localization of Zona Pellucida Receptors on Live Sperm by Fluorophore-Conjugated Solubilized Zona Pellucida Proteins Heather R. Burkin, Ana Paula Alves-Vieira, and David J. Miller Detection of Human Papillomavirus DNA in Sperm Using Polymerase Chain Reaction Olufemi A. Olatunbosun, Allison M. Case, and Harry G. Deneer Spermatogonial Stem Cells Karim Nayernia, Manyu Li, and Wolfgang Engel Molecular and Cellular Approaches for Establishment of an In Vitro Spermatogenesis System Karim Nayernia and Wolfgang Engel Acrosin Activity Assay for the Evaluation of Mammalian Sperm Acrosome Reaction Yehudit Lax, Sara Rubinstein, and Haim Breitbart Isolation of Sea Urchin Sperm Plasma Membranes Kathryn J. Mengerink and Victor D. Vacquier 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 Heide Schatten and Amitabha Chakrabarti Microscopic Techniques for Studying Sperm-Oocyte Interaction During Fertilization and Early Embryo Development Mark W. Tengowski Immunocytochemical Staining of the Metaphase II Spindle in Mammalian Oocytes Steve Mullen and John K. Critser Spindle Imaging in Living Mammalian Oocytes Using the Liquid-Crystal Polarized Light Microscope and Its Practical Use Wei-Hua Wang, Jimmy Gill, and Cathy Boutin In Vitro Maturation and Fertilization of Pig Oocytes Heng-Yu Fan and Qing-Yuan Sun In Vitro Maturation and In Vitro Fertilization of Human Oocytes Wei-Hua Wang and Jimmy Gill In Vitro Maturation and Fertilization of Canine Oocytes Karine Reynaud, Marie Saint-Dizier, and Sylvie Chastant-Maillard Detection and Measurement of Membrane-Bound Protein Tyrosine Kinases in the Zebrafish Egg Wenjun Wu and William H. Kinsey A Single-Cell Method for Estimation of Protein Tyrosine Kinases in the Zebrafish Egg Elinor MacGregor and William H. Kinsey Activity of MAPK/p90rsk During Fertilization in Mice, Rats, and Pigs Qing-Yuan Sun and Heng-Yu Fan Index