Germ Cell Protocols -

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Résumé :
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...

Biographie:
Cryopreservation of Semen of the Salmonidae with Special Reference to Large-Scale Fertilization.- Fertilization and Sperm Chemotaxis in Ascidians.- Assays for Vertebrate Sperm Chemotaxis.- Stallion Spermatozoa Viability.- Visualization of Sperm Accessory Structures in the Mammalian Spermatids, Spermatozoa, and Zygotes by Immunofluorescence, Confocal, and Immunoelectron Microscopy.- Localization of Zona Pellucida Receptors on Live Sperm by Fluorophore-Conjugated Solubilized Zona Pellucida Proteins.- Detection of Human Papillomavirus DNA in Sperm Using Polymerase Chain Reaction.- Spermatogonial Stem Cells.- Molecular and Cellular Approaches for Establishment of an In Vitro Spermatogenesis System.- Acrosin Activity Assay for the Evaluation of Mammalian Sperm Acrosome Reaction.- Isolation of Sea Urchin Sperm Plasma Membranes.- 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.- Microscopic Techniques for Studying Sperm-Oocyte Interaction During Fertilization and Early Embryonic Development.- Immunocytochemical Staining of the Metaphase II Spindle in Mammalian Oocytes.- Spindle Imaging in Living Mammalian Oocytes Using the Liquid-Crystal Polarized Light Microscope and Its Practical Use.- In Vitro Maturation and Fertilization of Pig Oocytes.- In Vitro Maturation and In Vitro Fertilization of Human Oocytes.- In Vitro Maturation and Fertilization of Canine Oocytes.- Detection and Measurement of Membrane-Bound Protein Tyrosine Kinases in the Zebrafish Egg.- Single-Cell Method for Estimation of Protein Tyrosine Kinases in the Zebrafish Egg.- Activity of MAPK/p90rsk During Fertilization inMice, Rats, and Pigs....

Sommaire:
we have selected those systems that have provided landmarks in advancing our knowledge on germ cells....