Cell Cycle Synchronization

This volume covers a broad range of cell types including cultured cell lines, primary cells, and various unicellular organisms such as fission yeast, budding yeast, parasite Leishmania amazonensis, and parasite Trypanosoma brucei. The chapters in this book are organized into four parts. Part One looks at a general overview of cell cycle control and synchronization. Part Two discusses techniques to synchronize mammalian cells to various cell cycle phases including mitotic sub-phases. Part Three covers synchronization of unicellular organisms and Part Four analyzes cell cycle progression. 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 thorough, Cell-Cycle Synchronization: Methods and Protocols is a valuable resource for both novice and expert scientists in this developing field.

​ This volume brings together a unique collection of protocols that cover standard, novel, and specialized techniques. Cell Cycle Oscillators: Methods and Protocols guides readers through recent progress in the field from both holistic and reductionist perspectives, providing the latest developments in molecular biology techniques, biochemistry, and computational analysis used for studying oscillatory networks. 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, Cell Cycle Oscillators: Methods and Protocols will serve as an invaluable reference to gain further insight into the complex and incompletely understood processes that are involved in the cell cycle and its regulation by oscillatory networks.

Cell Synchrony: Studies in Biosynthetic Regulation focuses on the processes, principles, applications, methodologies, and approaches involved in biosynthetic regulation, including chromosome replication, cellular biosynthesis, and enzyme synthesis. The selection first takes a look at the theory of synchronous cultures, synchronous replication of the chromosome in Bacillus subtilis, and chromosome replication in Escherichia coli. Discussions focus on attempts to characterize the physical state of replicating DNA, cell division and the regulation of chromosome replication, regulation of sequential chromosome synthesis, synchrony of chromosome replication, and determination of the cell doubling-time distribution function from the decay of synchronization. The text then examines nuclear events in synchronously dividing yeast cultures, periodic enzyme synthesis in synchronous cultures of yeast, and light-induced synchronous sporulation of a myxomycete. Topics include periodicity of enzyme synthesis, use of synchronous cultures for studying cellular biosynthesis, and cytological observations on synchronously dividing cultures. The book explores the effects of temperature on the mitotic cycle of normal and synchronized mammalian cells; behavior of sulfhydryl groups in synchronous division; and morphogenetic and macromolecular aspects of synchronized Tetrahymena. The selection is highly recommended for researchers interested in biosynthetic regulation.

The fundamental question of how cells grow and divide has perplexed biologists since the development of the cell theory in the mid-19th century, when it was recognized by Virchow and others that “all cells come from cells.” In recent years, considerable effort has been applied to the identification of the basic molecules and mechanisms that regulate the cell cycle in a number of different organisms. Such studies have led to the elucidation of the central paradigms that underpin eukaryotic cell cycle control, for which Lee Hartwell, Tim Hunt, and Paul Nurse were jointly awarded the Nobel Prize for Medicine and Physiology in 2001 in recognition of their seminal contributions to this field. The importance of understanding the fundamental mechanisms that modulate cell division has been reiterated by relatively recent discoveries of links between cell cycle control and DNA repair, growth, cellular metabolism, development, and cell death. This new phase of integrated cell cycle research provides further challenges and opportunities to the biological and medical worlds in applying these basic concepts to understanding the etiology of cancer and other proliferative diseases.

Cell Growth and Cell Division is a collection of papers dealing with the biochemical and cytological aspects of cell development and changes in bacterial, plant, and animal systems. One paper discusses studies on the nuclear and cytoplasmic growth of ten different strains of the genus Blepharisma, in which different types of nutrition at high and low temperatures alter the species to the extent that they became morphologically indistinguishable. The paper describes the onset of death at high and low temperatures as being preceded by a decrease in the size of the cytoplasm and a corresponding decrease in the size of the macronucleus. The moribund organisms, still possessing structure, are motionless with no distinguishable macronuclear materials. Another paper presents the response of meiotic and mitotic cells to azaguanine, chloramphenicol, ethionine, and 5-methyltryptophan. The paper describes the failure of spindle action, arrest of second division, inhibition of cytokinesis, aberrant wall synthesis, and alterations in chromosome morphology in meiosis cells. In the case of mitosis, a single enzyme—thymidine phosphorylase—shows that reagents which inhibit protein synthesis also inhibit the appearance of that enzyme if the reagent is applied one day before it normally appears. Other papers discuss control mechanisms for chromosome reproduction in the cell cycle, as well as the force of cleavage of the dividing sea urchin egg. The collection can prove valuable for bio-chemists, cellular biologists, micro-biologists, and developmental biologists.

During their lifetime, especially when growing and dividing, cells go through various steps of the cell cycle. Knowledge of the individual steps of the cell cycle will help us understand the development of a variety of diseases better, including cancer, and also to design new drugs against it. New techniques for studying the molecular basis of these processes have recently been developed and are described in detail in this manual. A glossary helps the reader to cope with the complex cell cycle terminology.

A collection of new reviews and protocols from leading experts in cell cycle regulation, Cell Cycle Control: Mechanisms and Protocols, Second Edition presents a comprehensive guide to recent technical and theoretical advancements in the field. Beginning with the overviews of various cell cycle regulations, this title presents the most current protocols and state-of-the-art techniques used to generate latest findings in cell cycle regulation, such as protocols to analyze cell cycle events and molecules. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Cell Cycle Control: Mechanisms and Protocols, Second Edition will be a valuable resource for a wide audience, ranging from the experienced cell cycle researchers looking for new approaches to the junior graduate students giving their first steps in cell cycle research.

Quantification of the proliferative characteristics of normal and malignant cells has been of interest to oncolo gists and cancer biologists for almost three decades. This interest stems from (a) the fact that cancer is a disease of uncontrolled proliferation, (b) the finding that many of the commonly used anticancer agents are preferentially toxic to cells that are actively proliferating, and (c) the observa tion that significant differences in proliferation characteristics exist between normal and malignant cells. Initially, cell cycle analysis was pursued enthusiastically in the hope of gener ating information useful for the development of rational cancer therapy strategies; for example, by allowing identi fication of rapidly proliferating tumors against which cell cycle-specific agents could be used with maximum effec tiveness and by allowing rational scheduling of cell cyc- specific therapeutic agents to maximize the therapeutic ratio. Unfortunately, several difficulties have prevented realiza tion of the early promise of cell cycle analysis: Proliferative patterns of the normal and malignant tissues have been found to be substantially more complex than originally an ticipated, and synchronization of human tumors has proved remarkably difficult. Human tumors of the same type have proved highly variable, and the cytokinetic tools available for cell cycle analysis have been labor intensive, as well as somewhat subjective and in many cases inapplicable to humans. However, the potential for substantially improved cancer therapy remains if more accurate cytokinetic infor mation about human malignancies and normal tissues can be obtained in a timely fashion.