This volume describes research methodologies and approaches used to study the Linker of Nucleoskeleton and Cytoskeleton (LINC) complex and its cellular functions. Chapters detail structural and biochemical analysis of LINC complexes, mechanical aspects of the LINC complex, analysis of the LINC complex in model systems and development, and LINC complex in mammalian tissue, organs, and disease. 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, The LINC Complex: Methods and Protocols aims to ensure successful results in the further study of this vital field.

LINC complexes span the nuclear envelope and connect the nucleoskeleton to the cytoskeleton. In 2012, our lab solved the first LINC complex structure, that of SUN domain of human SUN2 bound with KASH1 or KASH2 peptides. In this project testes-specific human SUN proteins (SUN3, SPAG4, and SUNS) were compared to ubiquitously-expressed SUN2. Secondly, fission and budding yeast LINC complexes differ from human ones and were analyzed as well. I was able to confirm SUN-KASH interaction in human and yeast. For structural analysis I explored various expression strategies. Fic-1 is a C. elegans Fic-domain protein with diverse cellular functions. As a subfamily III Fic enzyme, Fic-1 may reveal valuable insights into Fic enzyme mechanisms from its structure. After trying different knowledge-informed constructs and crystal optimization, small Fic-1 crystals were obtained, which diffracted X-rays to ~ 7 [angstroms]. With modest additional effort diffraction-quality crystals should be achievable.

This volume details established experimental protocols on the isolation, characterization, and detection of extracellular matrix (ECM) molecules as well as methods to study the activity and role of ECM components on various biological functions, the formation of exosomes, and tissue surrogates. Chapters display a variety of protocols ranging from biochemical, cell, and molecular biology assays to complex tissue imaging techniques and in vivo models to elucidate the role of the extracellular matrix. 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, The Extracellular Matrix: Methods and Protocols aims to provide protocols and applications that can be used by both Ph.D. students and senior researchers to advance research in the field of extracellular matrix biology.

The nucleus is the hallmark of the eukaryotic cell. It contains most of the genetic material and it separates the processes of replication and transcription from that of translation. Communication between the nucleus and the cytoplasm occurs mostly through openings in the nuclear envelope composed of nuclear pore complexes. These massive assemblies allow for regulated transport of macromolecules across the barrier that is the nuclear envelope. However, another means of communication between the nucleus and the cytoskeleton has been characterized: linkers of the nucleoskeleton and cytoskeleton, or LINC, complexes mechanically connect the nucleus with its surroundings allowing for nuclear anchorage and nuclear movement during development and chromosome movement during meiosis. At the heart of LINC complexes are inner nuclear membrane resident SUN proteins and outer nuclear membrane resident KASH proteins. In this thesis we structurally characterize the human SUN2-KASH1 and KASH2 complexes and provide the molecular basis for their interaction. The solved structures suggest plausible models for high-order of LINC complex assembly as well as LINC complex mediated spacing between the inner and outer nuclear membrane. Questions about how these complexes are regulated also arise from the structure. TorsinA is an AAA+ ATPase suggested to play a role in LINC complex regulation. Analysis of Torsin's binding partners LAP1 and LULL1 show that they are catalytically inactive AAA+ ATPases. We characterize the complex and show by EM that they form ring akin to other AAA+ ATPases. With these studies we provide the first structural analysis of LINC complexes and Torsin ATPases and also provide biochemical tools for the study of LINC complex regulation.

The genetic, molecular, and cellular mechanisms of neural development are essential for understanding evolution and disorders of neural systems. Recent advances in genetic, molecular, and cell biological methods have generated a massive increase in new information, but there is a paucity of comprehensive and up-to-date syntheses, references, and historical perspectives on this important subject. The Comprehensive Developmental Neuroscience series is designed to fill this gap, offering the most thorough coverage of this field on the market today and addressing all aspects of how the nervous system and its components develop. Particular attention is paid to the effects of abnormal development and on new psychiatric/neurological treatments being developed based on our increased understanding of developmental mechanisms. Each volume in the series consists of review style articles that average 15-20pp and feature numerous illustrations and full references. Volume 2 offers 56 high level articles devoted mainly to Formation of Axons and Dendrites, Migration, Synaptogenesis, Developmental Sequences in the Maturation of Intrinsic and Synapse Driven Patterns. - Series offers 144 articles for 2904 full color pages addressing ways in which the nervous system and its components develop - Features leading experts in various subfields as Section Editors and article Authors - All articles peer reviewed by Section Editors to ensure accuracy, thoroughness, and scholarship - Volume 2 sections include coverage of mechanisms which regulate: the formation of axons and dendrites, cell migration, synapse formation and maintenance during development, and neural activity, from cell-intrinsic maturation to early correlated patterns of activity

Progress in Molecular Biology and Translational Science provides a forum for discussion of new discoveries, approaches, and ideas in molecular biology. It contains contributions from leaders in their fields and abundant references. Volume 126 features in-depth reviews that focus on the tools required to investigate mechanotransduction. Additional chapters focus on how we can use these tools to answer fundamental questions about the interaction of physical forces with cell biology, morphogenesis, and function of mature structures. Chapters in the volume are authored by a unique combination of cell biologists and engineers, providing a range of perspectives on mechanotransduction. - Provides a unique combination of perspectives from biologists and engineers - Engaging to people of many training backgrounds

"Nuclear envelope (NE) defects have been linked to cancer biology since the mid-1800s, but it was not until the last few years that we have begun to understand these historical links and to realize that there are myriad ways that the NE impacts on tumorigenesis. The NE is a complex double membrane system that encloses the genome while providing structural support through the intermediate filament lamin polymer and regulating protein/ mRNA trafficking and signaling between the nucleus and cytoplasm via the nuclear pore complexes (NPCs). These functions already provide some mechanisms for NE influences on cancer biology but work in the past few years has elucidated many others. Lamins and many recently identified NE transmembrane proteins (NETs) have been now shown to function in DNA repair, regulation of cell cycle and signaling, apoptosis, cell migration in metastasis and nuclear architecture and morphology. This volume presents a comprehensive overview of the wide range of functions recently identified for NE proteins and their relevance in cancer biology, providing molecular mechanisms and evidence of their value as prognostic and diagnostic markers and suggesting new avenues for the treatment of cancer. Indeed some of these recent links are already yielding promising therapies, such as the current clinical trial of selective inhibitors of the nuclear export factor exportin in certain types of leukemia, melanoma and kidney cancer."

The delightful debut novel from the acclaimed author of "Southern Living" is finally available in paperback. A husband and dad discovers the unexpected responsibilities, pitfalls, and joys of being a full-time homemaker when his wife lands her dream job.

A remarkable debut mystery from the award-winning author of the 2003 St. Martin's Press/Private Eye Writers of America Prize for Best First Private Eye Novel. Michael Koryta's Tonight I Said Goodbye marks the emergence of a stunning new voice in crime fiction. With its edge-of-your-seat pacing, finely drawn characters, and rock-solid prose, Tonight I Said Goodbye would seem to be the work of a grizzled pro; the fact that the author is just twenty-one years old makes it all the more amazing. Investigator Wayne Weston is found dead of an apparent suicide in his home in an upscale Cleveland suburb, and his wife and six-year-old daughter are missing. Weston's father insists that private investigators Lincoln Perry and Joe Pritchard take the case to exonerate his son and find his granddaughter and daughter-in-law. As they begin to work they discover there is much more to the situation than has been described in the prevalent media reports. There are rumors of gambling debts and extortion, and a group of Russians with ties to organized crime who don't appreciate being investigated--a point they make clear with baseball bats. With some assistance from newspaper reporter Amy Ambrose, Perry and Pritchard believe they are making swift progress. But then they are warned off the investigation by a millionaire real estate tycoon and the FBI. Just when they feel they are closing in on a possible source of answers, another murder forces them to change direction in the case. Perry travels to a resort town in South Carolina and there he finds more than one game being played, and all of them are deadly. The stakes quickly become very personal for Perry, and it's clear that there will be no walking away from this case. In a debut that has already garnered praise from some of today's top writers, Michael Koryta immediately establishes himself as a standard bearer for the next generation of crime writers. Tonight I Said Goodbye is a 2005 Edgar Award Nominee for Best First Novel.