Germ Cell Protocols, Softcover reprint of hardcover 1st ed. 2004 Volume 2: Molecular Embryo Analysis, Live Imaging, Transgenesis, and Cloning Methods in Molecular Biology Series, Vol. 254

The study of germ cells has undergone enormous advances in recent years and has entered into an explosive phase of new discoveries with the introd- tion of transgenic technologies and nuclear cloning. Basic knowledge and te- niques 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 st- ies of fertility and infertility in mammals including humans. 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 asc- ian eggs, now benefits cloning efficiency in farm and domestic animals. Gene manipulations and molecular methods have added to the possibilities of p- ducing live offspring with enormous biomedical, ecological, and economic implications. All sexually reproducing organisms produce primordial germ cells, a small population of cells that differentiate into gametes of either sex that carry to- potency, an ability to develop into an entire new organism. The two volumes on germ cells combine techniques in a variety of different systems and have selected those systems that have provided landmarks in advancing our kno- edge on germ cells.

Activity of the Germline-Specific Oct4-GFP Transgene in Normal and Clone Mouse Embryos.- Analysis of Germline Chromatin Silencing by Double-Stranded RNA-Mediated Interference (RNAi) in Caenorhabditis elegans.- cDNA Subtraction and Cloning in the Field of Trophoblast/Placental Development.- PCR-Based Cloning and Differential Screening of RNAs from Xenopus Primordial Germ Cells.- Differentiating Embryonic Stem Cells into Embryoid Bodies.- The Derivation of Murine Embryonic Stem and Embryonic Germ Cells by Selective Ablation.- Using Multiphoton Excitation to Explore the Murky Depths of Developing Embryos.- Dual-Channel Confocal Ratioing of Calcium Dynamics in Living Eggs and Oocytes.- A Method for Producing Cloned Pigs by Using Somatic Cells as Donors.- Pronuclear Microinjection of Mouse Zygotes.- Parthenogenesis and Nuclear Transfer in Rabbit Oocytes.- Nuclear Transfer in Cattle.- In Vitro Fertilization and Embryo Transfer in Felids.- Production of Chimeric Chickens.- Production of Haploid and Diploid Androgenetic Zebrafish.- Construction and Detection of Fluorescent, Germline Transgenic Zebrafish.- Production of Zebrafish Germline Chimeras from Cultured Cells.- Studying Nuclear Reprogramming with Cell Hybrids.- Cytoplasmic Transfer in Mammalian Eggs and Embryos.- Conventional Technologies for Cryopreservation of Human Oocytes and Embryos.- Vitrifying and Warming of Human Oocytes, Embryos, and Blastocysts.- Microfluidic Removal of Cumulus Cells from Mammalian Zygotes.- Mammalian Embryo Culture in a Microfluidic Device.- Application of Alginate Gels to the Study of Mammalian Development.- Using TUNEL in Combination with an Active Caspase-3 Immunoassay to Identify Cells Undergoing Apoptosis in Preimplantation Mammalian Embryos.

Includes supplementary material: sn.pub/extras