Germ Cell Protocols

1. Front Matter

   Pages i-xiv

   PDF

2. Cryopreservation of Semen of the Salmonidae with Special Reference to Large-Scale Fertilization

   • Franz Lahnsteiner

   Pages 1-12

3. Fertilization and Sperm Chemotaxis in Ascidians

   • Manabu Yoshida

   Pages 13-25

4. Assays for Vertebrate Sperm Chemotaxis

   • Hitoshi Sugiyama, Bader Al-Anzi, Robert W. McGaughey, Douglas E. Chandler

   Pages 27-48

5. Stallion Spermatozoa Viability

   • Katrina Merkies, Mary M. Buhr

   Pages 49-58

6. Visualization of Sperm Accessory Structures in the Mammalian Spermatids, Spermatozoa, and Zygotes by Immunofluorescence, Confocal, and Immunoelectron Microscopy

   • Peter Sutovsky

   Pages 59-77

7. Localization of Zona Pellucida Receptors on Live Sperm by Fluorophore-Conjugated Solubilized Zona Pellucida Proteins

   • Heather R. Burkin, Ana Paula Alves-Vieira, David J. Miller

   Pages 79-94

8. Detection of Human Papillomavirus DNA in Sperm Using Polymerase Chain Reaction

   • Olufemi A. Olatunbosun, Allison M. Case, Harry G. Deneer

   Pages 95-104

9. Spermatogonial Stem Cells

   • Karim Nayernia, Manyu Li, Wolfgang Engel

   Pages 105-120

10. Molecular and Cellular Approaches for Establishment of an In Vitro Spermatogenesis System

    • Karim Nayernia, Wolfgang Engel

    Pages 121-133

11. Acrosin Activity Assay for the Evaluation of Mammalian Sperm Acrosome Reaction

    • Yehudit Lax, Sara Rubinstein, Haim Breitbart

    Pages 135-140

12. Isolation of Sea Urchin Sperm Plasma Membranes

    • Kathryn J. Mengerink, Victor D. Vacquier

    Pages 141-150

13. 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, Amitabha Chakrabarti

    Pages 151-164

14. Microscopic Techniques for Studying Sperm-Oocyte Interaction During Fertilization and Early Embryonic Development

    • Mark W. Tengowski

    Pages 165-199

15. Immunocytochemical Staining of the Metaphase II Spindle in Mammalian Oocytes

    • Steven F. Mullen, John K. Critser

    Pages 201-213

16. Spindle Imaging in Living Mammalian Oocytes Using the Liquid-Crystal Polarized Light Microscope and Its Practical Use

    • Wei-Hua Wang, Jimmy Gill, Cathy Boutin

    Pages 215-225

17. In Vitro Maturation and Fertilization of Pig Oocytes

    • Heng-Yu Fan, Qing-Yuan Sun

    Pages 227-233

18. In Vitro Maturation and In Vitro Fertilization of Human Oocytes

    • Wei-Hua Wang, Jimmy Gill

    Pages 235-253

19. In Vitro Maturation and Fertilization of Canine Oocytes

    • Karine Reynaud, Marie Saint-Dizier, Sylvie Chastant-Maillard

    Pages 255-272

20. Detection and Measurement of Membrane-Bound Protein Tyrosine Kinases in the Zebrafish Egg

    • Wenjun Wu, William H. Kinsey

    Pages 273-283

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.

• Activation
• DNA
• cryopreservation
• electron microscopy
• genes
• mammalian oocytes
• reproducible techniques

• Department of Veterinary Pathobiology, University of Missouri—Columbia, Columbia

  Heide Schatten

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