This book serves as an introduction to targeted genome editing, beginning with the background of this rapidly developing field and methods for generation of engineered nucleases. Applications of genome editing tools are then described in detail, in iPS cells and diverse organisms such as mice, rats, marine invertebrates, fish, frogs, and plants. Tools that are mentioned include zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and CRISPR/Cas9, all of which have received much attention in recent years as breakthrough technologies. Genome editing with engineered nucleases allows us to precisely change the target genome of living cells and is a powerful way to control functional genes. It is feasible in almost all organisms ranging from bacteria to plants and animals, as well as in cultured cells such as ES and iPS cells. Various genome modifications have proven successful, including gene knockout and knock-in experiments with targeting vectors and chromosomal editing. Genome editing technologies hold great promise for the future, for example in biomedical research, clinical medicine, and generation of crops and livestock with desirable traits. A wide range of readers will find this book interesting, and with its focus on applications in a variety of organisms and cells, the book will be valuable for life scientists in all fields.

Drawing on state-of-the-art cellular and molecular techniques as well as new and sophisticated imaging and information technologies, this comprehensive, three-volume collection of cutting-edge protocols provides readily reproducible methods for studying and analyzing the events of embryonic development. Volume 1 (ISBN: 089603-574-3) contains techniques for establishing and characterizing several widely used experimental model systems, for the study of developmental patterns and morphogenesis, and for the examination of embryo structure and function. There are also step-by-step methods for the analaysis of cell lineage, the production and use of chimeras, and the experimental and molecular manipulation of embryos, including the application of viral vectors. Volume 2 (ISBN: 0-89603-575-1) describes state-of-the-art methods for the study of organogenesis, the analysis of abnormal development and teratology, the screening and mapping of novel genes and mutations, and the application of transgenesis, including the production of transgenic animals and gene knockouts. No less innovative, volume 3 (ISBN: 0-89603-576-X) introduces powerful techniques for the manipulation of developmental gene expression and function, the analysis of gene expression, the characterization of tissue morphogenesis and development, the in vitro study of differentiation and development, and the genetic analysis of developmental models of diseases. Highly practical and richly annotated, the three volumes of Developmental Biology Protocols describe multiple experimental systems and details techniques adopted from the broadest array of biomedical disciplines.

Drawing on state-of-the-art cellular and molecular techniques as well as new and sophisticated imaging and information technologies, this comprehensive, three-volume collection of cutting-edge protocols provides readily reproducible methods for studying and analyzing the events of embryonic development. Volume 1 (ISBN: 089603-574-3) contains techniques for establishing and characterizing several widely used experimental model systems, for the study of developmental patterns and morphogenesis, and for the examination of embryo structure and function. There are also step-by-step methods for the analaysis of cell lineage, the production and use of chimeras, and the experimental and molecular manipulation of embryos, including the application of viral vectors. Volume 2 (ISBN: 0-89603-575-1) describes state-of-the-art methods for the study of organogenesis, the analysis of abnormal development and teratology, the screening and mapping of novel genes and mutations, and the application of transgenesis, including the production of transgenic animals and gene knockouts. No less innovative, Volume 3 (ISBN: 0-89603-576-X) introduces powerful techniques for the manipulation of developmental gene expression and function, the analysis of gene expression, the characterization of tissue morphogenesis and development, the in vitro study of differentiation and development, and the genetic analysis of developmental models of diseases. Highly practical and richly annotated, the three volumes of Developmental Biology Protocols describe multiple experimental systems and details techniques adopted from the broadest array of biomedical disciplines.

Cheryl S. Watson University o/Texas Medical Branch Cellular steroid action has been thoroughly studied in the nuclear compartment. However, nuclear steroid receptor mechanisms have been unable to explain some of the rapid activities of steroids, partiCUlarly those which occur in a time frame of seconds to minutes reviewed in (1;2)]. Based on these and other considerations, an alternative membrane-associated receptor form was long ago proposed to exist (3). Others interpret the location of the steroid receptors mediating these rapid effects as peri membrane or cytoplasmic. New experimental tools have been brought to bear on the topic of receptors for steroids which mediate non-genomic actions, and thus investigative activity and focus regarding this type of steroid receptor has recently increased significantly. However, there may be multiple answers to the question "how do steroids mediate rapid nongenomic effects?" Steroid actions initiated at the cell membrane can impinge on important phases in the lifespan of a cell: proliferation, migration, differentiation, and release of hormones or neurotransmitters functioning as signals to other cells."

This book challenges the assumption that morphological data are inherently unsuitable for phylogeny reconstruction, argues that both molecular and morphological phylogenies should play a major role in systematics, and provides the most comprehensive review of the comparative anatomy, homologies and evolution of the head, neck, pectoral and upper limb muscles of primates. Chapters 1 and 2 provide an introduction to the main aims and methodology of the book. Chapters 3 and 4 and Appendices I and II present the data obtained from dissections of the head, neck, pectoral and upper limb muscles of representative members of all the major primate groups including modern humans, and compare these data with the information available in the literature. Appendices I and II provide detailed textual (attachments, innervation, function, variations and synonyms) and visual (high quality photographs) information about each muscle for the primate taxa included in the cladistic study of Chapter 3, thus providing the first comprehensive and up to date overview of the comparative anatomy of the head, neck, pectoral and upper limb muscles of primates. The most parsimonious tree obtained from the cladistic analysis of 166 head, neck, pectoral and upper limb muscle characters in 18 primate genera, and in representatives of the Scandentia, Dermoptera and Rodentia, is fully congruent with the evolutionary molecular tree of Primates, thus supporting the idea that muscle characters are particularly useful to infer phylogenies. The combined anatomical materials provided in this book point out that modern humans have fewer head, neck, pectoral and upper limb muscles than most other living primates, but are consistent with the proposal that facial and vocal communication and specialized thumb movements have probably played an important role in recent human evolution. This book will be of interest to primatologists, comparative anatomists, functional morphologists, zoologists, physical anthropologists, and systematicians, as well as to medical students, physicians and researchers interested in understanding the origin, evolution, homology and variations of the muscles of modern humans. Contains 132 color plates.

Epithelial phenotype is a dynamic stage of differentiation that can be modulated during several physiological or pathological events. The rapid conversion to a mesenchymal-like phenotype is called an epithelial-mesenchymal transition (EMT).

The Rise and Fall of Epithelial Phenotype is the first book to comprehensively introduce the concept of EMT. The first part of this volume describes main examples and models and explains their physiological relevance. These examples include hydra morphogenesis, gastrulation in mouse, drosophila and sea urchin, as well as neural crest cell migration and heart morphogenesis in vertebrates. Part two reviews in detail, specific EMT molecular pathways covering extracellular induction, transduction and transcription response and modulation of cell-cell adhesion structures. It emphasizes new specific pathways with potential medical applications. EMTs can also be linked to pathological events such as wound healing and cancer progression, as detailed in this section of the book.

The purpose of this third edition of Developmental and Reproductive Toxicology is to provide a practical guide to developmental and reproductive toxicology in a regulatory environment. In addition to a comprehensive update of current chapters, the third edition been revised to reflect recent changes in the field. It contains new chapters that reflect emerging topics of interest, including testing of biologics (including vaccines), nonhuman primates as nonclinical models, developmental immunotoxicity testing, in vitro assays (such as use of zebrafish and stem cells, as well as high throughput screening), in silico systems modelling, evaluating mechanisms of reproductive toxicity, in-depth coverage of neurobehavioral testing, and testing under the EU's REACH regulations, as well as updated chapters on nonclinical juvenile toxicity testing, endocrine disruptor screening, and on functional and computational genomics. The study of hazard and risk associated with exposure to toxicants during prenatal development has been expanded in recent years to include effects on development until the time of puberty. Concern over the adverse effects of chemical or physical agents on the reproductive processes of both sexes has increased, and progress has been made in identifying the causes and mechanisms eliciting congenital defects and determining the genetic, epigenetic, and environmental factors involved. This book provides up-to-date guidance on the use and interpretation of the newest research techniques in developmental and reproductive toxicology, as well as the more traditional approaches. Developmental and Reproductive Toxicology, Third Edition: Contains valuable insights gained from hands-on experience, together with a critical evaluation of current testing strategies. Includes guidance for the design, conduct, and interpretation of tests in all areas of developmental and reproductive toxicity. Contains reprinted guidelines from major regulatory agencies, as well as terminology for description of developmental abnormalities in laboratory animals, for easy reference. Provides guidance for planning and conducting preclinical toxicity studies and follow-up studies, and interpreting their results in a regulatory environment. This book continues to be the ideal practical reference for developmental and reproductive toxicologists who perform research in industry, government, and academia and for anyone who intends to enter these research areas.

The electric utility industry and its stakeholders in the.United States appear to be at a critical juncture in time. Powerful forces of global proportions are propelling the industry instinctively and in a secular fashion towards restructuring. That the industry will change is a fait accomplii. The nature and timing of the change is still a matter of intense debate, however. Because of the evolution of the industry into its present-day form, i.e. regulated local monopolies in their designated franchise service territories, the relative roles and expectations of various institutions would have to change to conform to the new state in the future. In either encouraging, or allowing this change to happen, society is essentially saying that future societal welfare would be better served by the changed structure contemplated. What that assumption translates into in more direct terms is that creation of future wealth would be better accomplished through redistribution of wealth today. Thoughtful individuals recognize the enormous responsibility placed upon the various entities empowered with jurisdiction over the timing and nature of the structural change. They are trying hard to bring analytical rigor to bear on the debate. One very critical element of this debate on restructuring is the issue of the treatment of transmission. The issue has been variously labeled transmission access, or pricing. Volumes have been written and spoken on this topic.

The nutritive endosperm of angiosperms is mankind 's most important source of food, livestock feed and industrial raw material. This book is the first comprehensive overview of the developmental and molecular biology of endosperm. The text covers cereal endosperm development from fertilization to maturity, including molecular and cell biology of the syncytial phase. It also goes into the cellularization process and cell fate specification of the embryo surrounding region cells, the basal transfer cells, the starchy endosperm and aleurone cells.

The innate immune response is a crucial component of early resistance to infection, and it is now revealing increasing levels of complexity. The ability to modify the genome in vivo, has facilitated understanding of complex interactions between leucocytes and other components of the immune system, and phenotype-driven strategies using chemical mutagenesis have placed another powerful weapon in the armamentarium. In, Leucocytes: Methods and Protocols provides detailed protocols and practical advice on a variety of modern approaches to the study of leucocytes and their products. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Leucocytes: Methods and Protocols seeks to aid scientists in further study into the immune system and leucocytes.

The human immune system is a complex network of tissues and organs dispersed throughout the body. Immunology, as one of the most rapidly evolving fields in bio-medical research, has to date covered the essential cellular and molecular events neces-sary for immune responses to occur. However, it has paid relatively little attention to important developmental processes underlying the formation of the tissues themselves that carry out immune responses in humans and other mammalians. In contrast to the thymus and bone marrow that are the sole tissues for generating mature leukocytes for antigen recognition and han-dling in humans and most mammalian species, the peripheral lymphoid tissues where adaptive immune responses are focused display broad tissue distribution and possess diverse archi-tectural characteristics. These organs develop prior to the individual s exposure to external antigens, and despite their similar functions, their varied appearances indicate a substantial complexity of tissue ontogeny. This volume presents a comprehensive overview of the developmental features of the major peripheral lymphoid organs, thus examining the connection between immunological functionality and structural characteristics utilizing a developmental approach, for an audience ranging from undergraduate students to senior researchers in immunology, histology and clinical medicine."

During the last 40 years, the study of the biological basis of aging has progressed tremendously, and it has now become an independent and respectable field of study and research. The essential cause of aging is molecular damage that slowly overwhelms cellular and organismic defense, repair and maintenance systems. In recent years, a wealth of highly sophisticated research has transformed this idea from a credible hypothesis not only to a major theory, but essentially to accepted knowledge. Aging at the Molecular Level examines the key elements in this transformation.
Bringing together contributions from an international team of authors, this volume will be of interest to graduates and postgraduates in the fields of medicine and nursing, researchers of different aspects of biogerontology and those in the pharmaceutical, cosmeceutical, nutraceutical and health-care industry.

In this issue of Current Topics in Microbiology and Immunology the authors present a unique range of examples of the regenerative response and the role of stem cells from the amphibian to human. It seems that all roads lead to cells that have the plasticity to become something else, not only in the amphibian but also in the mammal.

This volume discusses recent advances in avian and reptilian biology that have caused this diverse field to re-emerge. The chapters in this book are divided into 4 parts: genomics and transcriptomics, genetic manipulation, stem cells, and new model systems. Part I details how to perform genomic and transcriptomic analyses in birds and reptiles; Part II highlights technological advancements in avian genetic manipulation; Part III focuses on methods to handle pluripotent cells; and part IV looks at the emerging models in avian and reptilian developmental biology. 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. Cutting-edge and comprehensive, Avian and Reptilian Developmental Biology: Methods and Protocols explores a var iety of approaches and different sauropsid models that will help facilitate communication and collaboration among researchers, which in turn will progress this field forward.

For anybody capable of an emotional response to it, any view of a developing organism should give birth to a feeling of amazement and even admiration, whether this development is seen directly, or in the form of a time lapse film, or even if mentally reconstructed from a series of static images. We ask ourselves how such seemingly primitive eggs or pieces of tissue, without any obvious intervention from outside, so regularly transform themselves into precisely constructed adult organisms. If we try to formulate what amazes us most of all about development, the answer will probably be that it is the internal capacity of developing organisms themselves to create new structures. How, then, can we satisfy our amazement in ways that are more or less reasonable, as well as scientifically valuable? This depends, first of all, on what position we choose to regard embryonic development as occupying among other structure creating processes, even including human activities. On the one hand, one might regard the development of organisms as a highly specialized class of processes, unique to themselves and alien to the general laws of nature, or at least not derivable from them and more akin to the deliberate acts of our own human behaviour. In that case our task would become reduced to a search for some specific 'instructions' for each next member of such a class. Whether in an overt or hidden form, some such ideology seems to dominate in present day developmental biology.

Redox Metabolism and Longevity Relationships in Animals and Plants focuses on the recent issues that have emerged in ageing research in both the animal and plant kingdoms. This volume reviews current concepts concerning cellular redox homeostatis and ageing in animals and plants, relationships to programmed cell death, the production of oxidants and dicarbonyls, the ways that different organisms perceive and respond to oxidative, nitration and glycation challenges, and how this might be intricately connected to ageing and lifespan.

The organizer area plays a central role in the formation of the embryonic axis and the central nervous system of all vertebrates including the human fetus. In The Vertebrate Organizer, outstanding molecular development biologists and embryologists report their latest approaches in this fascinating research area using different vertebrate model organisms. The presented data is of central importance for the understanding of early human embryogenesis.

... an adult poet is simply an individual in a state of arrested development-in brief, a sort of moron. Just as all of us, in utero, pass through a stage in which we are tadpoles, ... so all of us pass through a state, in our nonage, when we are poets. A youth of seventeen who is not a poet is simply a donkey: his development has been arrested even anterior to that of the tadpole. But a man of fifty who still writes poetry is either an unfortunate who has never developed, intellectually, beyond his teens, or a conscious buffoon who pretends to be something he isn't-something far younger and juicier than he actually is. -H. 1. Mencken, High and Ghostly Matters, Prejudices: Fourth Series (1924) Where would evolution be, Without this thing, heterochrony? -M. L. McKinney (1987) One of the joys of working in a renascent field is that it is actually possible to keep up with the literature. So it is with mixed emotions that we heterochronists (even larval forms like myself) view the recent "veritable explosion of interest in heterochrony" (in Gould's words in this volume). On the positive side, it is ob viously necessary and desirable to extend and expand the inquiry; but one regrets that already we are beginning to talk past, lose track of, and even ignore each other as we carve out individual interests."

This book provides an overview of the role and function of regulatory RNAs that lack protein-coding potential in key reproductive tissues. This includes the role of small interfering RNAs (siRNAs), microRNAs (miRNAs), PIWI-interacting RNAs (piRNAs), small nucleolar RNAs (snoRNAs) and long non-coding RNAs (lncRNAs). Through clear, detailed and comprehensive debate, international leading experts discuss the role these novel regulators in normal development of sexual dimorphisms, including the differentiation of ovaries and testes, the genital tract including prostate, epididymis and uterus, as well as mammary glands. In addition, particular attention is paid on their role in pathophysiological processes within the reproductive tract. The power of next generation sequencing has proved to be an invaluable tool to discover new non-coding RNAs. While the identification of non-coding RNA is relatively easy, analysing their function represents still a challenge today. In this book, authors present historical and conceptual background information, highlight the ways in which non-coding RNAs function is analysed and present their vision of the future research in their key research area.

In 1964, the Fertilization and Gamete Physiology Research Training Program (FERGAP) was established at the Marine Biological Laboratories, Woods Hole, Massachusetts. Over the course of the next 12 years, under the directorship of Dr. Charles B. Metz, FERGAP brought together, trained, and inspired a generation of students in reproductive biology from all over the world. As students of C. B. Metz and as FERGAP trainees, we would like to dedicate this collected work on comparative mammalian fertilization to our teacher and mentor, Dr. Charles B. Metz. Like a number of authors contributing to this volume, we have been struck by the significant impact that C. B. Metz and FERGAP had on the development of students of reproductive biology. Applying both the classical and molecular techniques of cell biology and immunology to problems of gamete biology, Dr. Metz emphasized a comparative and analytical approach that was reflected in his own research on fertilization in Paramecia, sea urchins, frogs, and mammals. It is hoped that this volume will serve to stimulate students to discover the myriad of fascinating research problems in gamete and reproductive biology. Bonnie S. Dunbar Michael G. O'Rand Houston, Texas Chapel Hill, North Carolina ix Contents Part I COMPARATIVE OVERVIEW OF MAMMALIAN GAMETES The Coevolution of Mammalian Gametes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 J. Michael Bedford I. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 2. Gamete Design . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 2.1. Monotremes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 . . . . . . . . . . . . . . 2.2. Marsupials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 . . . . . . . . . . . . . . . 2.3. Eutherians . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 3. Gamete Maturation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 . . . . . . . . . . . . . . . 3 .1. Oocyte Maturation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 . . . . . . . . . . . . . 3.2. Sperm Maturation in the Male . . . . . . . . . . . . . . . . . . . . . . . 15 . . . . . . . . . .

This book presents a timely collection of reviews by experts in periconception and fertilization. The book provides a good introduction for those new to the field as well as those who have worked in this field for some time, but not have been able to keep up-to-date with recent advances on the topic. The book reviews the current knowledge in this field, then focuses its attention on short-term and long-term effects of the periconception period, both from a physiological and medical perspective. Finally the volume covers the examination of the potential mechanisms involved in controlling periconception period. From the early days of discovery about the basics of the fertilization process, scientists have known that early events happening during conception play a major part in the creation of new offspring. However, until nearly a decade ago we thought these events were just concerned with the conception and nothing to do with lifelong health and welfare, or wellbeing of the offspring. Early indications from IVF in sheep and cattle suggested that the embryo culture conditions employed in the laboratory could result in unusually large and unhealthy offspring. These observations sounded alarm bells and stimulated research into the impacts of in vivo and in vitro conditions on the health of embryos and offspring. This book provides a summary of the state-of-the-art research being done to more fully understand the importance of the periconception environment.

The book is divided into three parts: Flower, Anther, and Ovule. The principal aim of this volume (along with the other 3 volumes in the series) is to summarize the classical and current concepts about flower generative organs, their structure and development, and about seed formation processes. The book contains ample material that can be employed in theoretical generalizations, in analyzing the distribution of features (or their uniqueness) and evolutionary transformations of structures. This offers vast possibilities for revising the existing and developing new classifications and concepts.