This volume details a wide range of methods, ranging from beginner through advanced, used to further study zebrafish and fish germline. Chapter guide readers through cultivating and manipulating germ cells, imaging of germline processes and the molecular analysis of their, protein, and RNA. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, application details for both the expert and non-expert reader, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Germline Development in the Zebrafish: Methods and Protocols aims to ensure successful results in the further study of this vital field.
Eggs of all animals contain mRNAs and proteins that are supplied to or deposited in the egg as it develops during oogenesis. These maternal gene products regulate all aspects of oocyte development, and an embryo fully relies on these maternal gene products for all aspects of its early development, including fertilization, transitions between meiotic and mitotic cell cycles, and activation of its own genome. Given the diverse processes required to produce a developmentally competent egg and embryo, it is not surprising that maternal gene products are not only essential for normal embryonic development but also for fertility. This review provides an overview of fundamental aspects of oocyte and early embryonic development and the interference and genetic approaches that have provided access to maternally regulated aspects of vertebrate development. Some of the pathways and molecules highlighted in this review, in particular, Bmps, Wnts, small GTPases, cytoskeletal components, and cell cycle regulators, are well known and are essential regulators of multiple aspects of animal development, including oogenesis, early embryogenesis, organogenesis, and reproductive fitness of the adult animal. Specific examples of developmental processes under maternal control and the essential proteins will be explored in each chapter, and where known conserved aspects or divergent roles for these maternal regulators of early vertebrate development will be discussed throughout this review. Table of Contents: Introduction / Oogenesis: From Germline Stem Cells to Germline Cysts / Oocyte Polarity and the Embryonic Axes: The Balbiani Body, an Ancient Oocyte Asymmetry / Preparing Developmentally Competent Eggs / Egg Activation / Blocking Polyspermy / Cleavage/ Mitosis: Going Multicellular / Maternal-Zygotic Transition / Reprogramming: Epigenetic Modifications and Zygotic Genome Activation / Dorsal-Ventral Axis Formation before Zygotic Genome Activation in Zebrafish and Frogs / Maternal TGF-β and the Dorsal-Ventral Embryonic Axis / Maternal Control After Zygotic Genome Activation / Compensation by Stable Maternal Proteins / Maternal Contributions to Germline Establishment or Maintenance / Perspective / Acknowledgments / References
This second edition provides updated and new chapters on selected genetic, molecular, biochemical, and cell biological techniques. Chapter’s guide readers through methods and principles on primordial germ cells and germline stem cells, however many of these principles can be applied to different types of adult stem cells. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Germline Stem Cells: Second Edition aims to present the new findings and techniques that have developed since the first edition.
Life history theory seeks to explain the evolution of the major features of life cycles by analyzing the ecological factors that shape age-specific schedules of growth, reproduction, and survival and by investigating the trade-offs that constrain the evolution of these traits. Although life history theory has made enormous progress in explaining the diversity of life history strategies among species, it traditionally ignores the underlying proximate mechanisms. This novel book argues that many fundamental problems in life history evolution, including the nature of trade-offs, can only be fully resolved if we begin to integrate information on developmental, physiological, and genetic mechanisms into the classical life history framework. Each chapter is written by an established or up-and-coming leader in their respective field; they not only represent the state of the art but also offer fresh perspectives for future research. The text is divided into 7 sections that cover basic concepts (Part 1), the mechanisms that affect different parts of the life cycle (growth, development, and maturation; reproduction; and aging and somatic maintenance) (Parts 2-4), life history plasticity (Part 5), life history integration and trade-offs (Part 6), and concludes with a synthesis chapter written by a prominent leader in the field and an editorial postscript (Part 7).
This volume of Methods in Cell Biology, the second of two parts on the subject of zebrafish, provides a comprehensive compendium of laboratory protocols and reviews covering all the new methods developed since 1999. - Details state-of-the art zebrafish protocols, delineating critical steps in the procedures as well as potential pitfalls - Illustrates many techiques in full-color - Summarizes the Zebrafish Genome Project
This volume of Methods in Cell Biology is the 3e, and provides comprehensive compendia of laboratory protocols and reviews covering all the new methods developed since 2004. This new volume on Disease Models and Chemical Screens, covers two rapidly emerging and compelling applications of the zebrafish. - Details state-of-the art zebrafish protocols, delineating critical steps in the procedures as well as potential pitfalls - This volume concentrates on Disease Models and Chemical Screens
In this comprehensive and cutting-edge book, leading experts explore the parameters that define germline stem cells and the mechanisms that regulate the cell behavior in order to better isolate, characterize and maintain them. The volume begins by providing protocols for germline stem cell identification and regulation in model organisms, and concludes with detailed chapters covering current techniques involving in vitro culture and the applications of the cells.
Authored by leading experts in the field, this book provides the first comprehensive overview of the mechanisms of early patterning and morphogenesis in zebrafish. It summarizes the current knowledge and the key questions for the next decade of research.
Innovations in molecular biology are allowing neuroscientists to study the brain with unprecedented resolution, from the level of single molecules to integrated gene circuits. Chief among these innovations is the CRISPR-Cas genome editing technology, which has the precision and scalability to tackle the complexity of the brain. This Colloque Médecine et Recherche has brought together experts from around the world that are applying genome editing to address important challenges in neuroscience, including basic biology in model organisms that has the power to reveal systems-level insight into how the nervous system develops and functions as well as research focused on understanding and treating human neurological disorders. This work was published by Saint Philip Street Press pursuant to a Creative Commons license permitting commercial use. All rights not granted by the work's license are retained by the author or authors.
The chapters in this volume of "Insights from Animal Reproduction" address several, particular hot topics in the field of reproduction. The book begins with a comprehensive overview of the cryopreservation of sheep-produced embryos. The following chapter revises the assisted reproductive techniques available for South American wild mammals. Chapter 3 presents the technical procedures necessary to produce transgenic goats. Chapter 4 provides a comprehensive revision of the major molecular determinants of litter size in prolific species. Chapter 5 examines the germ cell determinant transmission, segregation, and function using the zebrafish as a model for germ cell specification in the embryo. Chapter 6 summarizes the current understanding of the molecular and cellular mechanisms regulating the early stages of folliculogenesis. Chapter 7 examines the sperm motility regulatory proteins as a tool to enhance sperm quality in cryopreservation processes. Chapter 8 discusses contemporary knowledge on the effects of extremely low frequency magnetic fields (ELF-MF) on male reproductive function in rodents. Chapter 9 highlights the importance of the cytogenetic evaluation in searching for causes of infertility of phenotypically normal animals, as well as individuals with an abnormal sex development. The last chapter provides evidence that other uterine diseases may be hidden behind the clinical diagnosis of pyometra that in some case may have a poor outcome.
