Download or read book Neuropeptide Signaling in Crustaceans Probed by Mass Spectrometry written by and published by . This book was released on 2016 with total page 564 pages. Available in PDF, EPUB and Kindle. Book excerpt: Neuropeptides are one of the most diverse classes of signaling molecules whose identities and functions are not yet fully understood. They have been implicated in the regulation of a wide range of physiological processes, including feeding-related and motivated behaviors, and also environmental adaptations. In this work, improved mass spectrometry-based analytical platforms were developed and applied to the crustacean systems to characterize signaling molecules. This dissertation begins with a review of mass spectrometry-based neuropeptide studies from both temporal- and spatial-domains. This review is then followed by several chapters detailing a few research projects related to the crustacean neuropeptidomic characterization and comparative analysis. The neuropeptidome of crayfish, Orconectes rusticus is characterized for the first time using mass spectrometry-based tools. In vivo microdialysis sampling technique offers the capability of direct sampling from extracellular space in a time-resolved manner. It is used to investigate the secreted neuropeptide and neurotransmitter content in Jonah crab, Cancer borealis, in this work. A new quantitation strategy using alternative mass spectrometry data acquisition approach is developed and applied for the first time to quantify neuropeptides. Coupling of this method with microdialysis enables the study of neuropeptide dynamics concurrent with different behaviors. Proof-of-principle experiments validating this approach have been carried out in Jonah crab, Cancer borealis to study feeding- and circadian rhythm-related neuropeptide changes using micoridialysis in a time-resolved manner. This permits a close correlation between behavioral and neurochemical changes, providing potential candidates for future validation of regulatory roles. In addition to providing spatial information, mass spectrometry imaging (MSI) technique enables the characterization of signaling molecules while preserving the temporal resolution. A novel MSI-based platform is developed by interfacing with microdialysis sample collection and is applied to the study of in vivo neuropeptide degradation profiles in an off-line ‘real-time’ fashion. This unique platform provides novel insights into neuropeptide inactivation/elimination process, which is an essential step to understand peptidegic signaling pathway. In addition to neuropeptides, this platform has been further modified to study secreted neurotransmitters, and small molecule metabolites that might interact with neuropeptides in crustacean for the first time. This work not only reports improvements upon neuropeptides identification and quantitation by developing improved analytical strategies, but also provides important evidence for the potential roles of several neuropeptides in regulating food intake and circadian rhythm behaviors. Novel platform that integrates MALDI-MSI with microdialysis in exploring neurochemical changes in dynamic biological events is also presented, which could be potentially applied to many other systems in future research. Collectively this dissertation research develops new analytical methods and improves our fundamental understanding of neuropeptide signaling.
Download or read book Neuropeptides in Crustaceans written by Chuanzi OuYang and published by . This book was released on 2016 with total page 341 pages. Available in PDF, EPUB and Kindle. Book excerpt: In this thesis, multifaceted biological mass spectrometry (MS) based methods were developed and employed to enable comprehensive characterization of neuropeptides. Neuropeptides encompass the largest and most diverse group of signaling molecules in the nervous system. They are necessary for the initiation and regulation of numerous physiological processes such as feeding, reproduction and development. Due to the complexity and inaccessibility of human systems, many invertebrate model nervous systems have been utilized as test bed for neuropeptidome research. Of these, the decapod crustacean nervous system provides an excellent test bed to facilitate technology development and investigate the biological functions of neuropeptides. A review of the current status and progress in the field of invertebrate neuropeptide is included in this thesis. Among various MS based technologies, we focused on developing new methodologies in mass spectral imaging (MSI) to provide high throughput tools for multiplexed distribution characterization. Current development of MSI and microdialysis based MS approaches have been summarized as the background of MSI related projects. To this rapidly expanding body of literature, the work presented here contributes insight into the possible functionality of certain peptides by identifying neuropeptides found to be differentially expressed in correlation with food intake and changes of environmental conditions. This comparative peptidomic study utilized direct tissue analysis, MSI and stable isotopic labeling to provide comprehensive information regarding to peptide expression level changes under different physiological states. In addition, sample preparation method was developed to detect trace level of neuropeptides from crustacean circulating fluid hemolymph. Furthermore, optimized sample preparation for imaging mass spectrometry was developed to greatly increase the coverage of peptidome and the accuracy of identification. Finally, using established techniques, novel neuropeptides were identified from the neuroendocrine organs in Dungeness crab and rock crab, and their distribution was also investigated. Collectively, this body of work extends the capability of mass spectrometry as a bioanalytical tool for neuropeptide analysis and advances the basic understanding of neuronal signaling.
Download or read book Probing Neuropeptidomics by Mass Spectrometry written by and published by . This book was released on 2012 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: In this thesis, an array of biological mass spectrometry (MS) based methods were developed and employed to enable the identification and functional analysis of neuropeptides in the crustacean nervous system. Neuropeptides encompass the largest and most diverse group of signaling molecules in the nervous system. They are necessary for the initiation and regulation of numerous physiological processes such as feeding, reproduction and development. However, the discovery and identification of these important signaling molecules presents unique challenges due to the vast complexity and minute quantities of endogenously expressed peptides. Further functional analysis of neuropeptides needs to be conducted for a comprehensive neuropeptidomic study. Using the decapod crustacean nervous system and associated neuroendocrine organs as experiment model, this dissertation research focuses on the development of an integrated analytical platform for large-scale identification and discovery of neuropeptides, as well as investigation of the functional roles of neuropeptides. The work presented here contributes insight into neuropeptidome of a commercially and scientifically important crustacean species, blue crab /italicCallinectes sapidus/italic and the possible functionality of certain peptides. A multifaceted mass spectrometry approach combining complementary sample preparation techniques, advanced separation strategies, alternative fragmentation methods, mass spectral imaging (MSI) and stable isotopic labeling was used for large-scale identification and to provide comprehensive information on peptide expression level changes under different physiological states. Several new analytical methodologies and tools have been developed to enable enhanced neuropeptidome coverage and improved /italicde novo/italic sequencing. A two dimensional liquid chromatography (2D-LC) method coupling reversed-phase liquid chromatography (RPLC) with nano-LC-ESI-Q-TOF MS/MS was used for neuronal tissue extract analysis and greatly improved the neuropeptidome coverage. A homemade capillary isoelectric focusing (cIEF) system was used for separation of the most acidic neuropeptide family Orcokinins in the sinus gland (SG). Additionally, electron transfer dissociation (ETD) was employed for the identification and sequencing of intact crustacean hyperglycemic hormone precursor related peptides (CPRPs), a group of multiply charged peptides in the SG. More than 150 neuropeptides including over 40 novel ones were identified in blue crab's nervous system. Furthermore, a comparative peptidomic study utilized mass spectral imaging (MSI), stable isotopic labeling and electrophysiological study was developed to provide comprehensive information about peptide expression level changes after food intake. Moreover, a MS-based platform was developed to study the differential degradation of neuropeptides by extracellular peptidases via monitoring degradation rate and degradation products. Collectively, this body of work extends the capability of mass spectrometry as a bioanalytical tool for neuropeptide analysis and advances the basic understanding of neuronal signaling.
Download or read book Discovery and Quantification of Crustacean Neuropeptides Using Mass Spectrometry and Informatics Approaches written by Nhu Vu and published by . This book was released on 2022 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: The study of endogenous peptides is important for discovering critical elements involved in many physiological processes and disorders. Neuropeptides are endogenous peptide hormones that regulate these processes by binding to receptors to initiate cellular signaling pathways. Discovery of neuropeptide function is an increasingly popular avenue for disease therapeutics research, resulting in increased need for improved neuropeptide characterization methods. Although mass spectrometry (MS) is well-suited for neuropeptide analysis due to its high sensitivity and capacity for untargeted, global neuropeptide measurements, there are still technical challenges that must be overcome. For example, neuropeptide sequences within a peptide family are highly homologous and therefore must be analyzed in their intact form to obtain high sequence coverage for confident identification and often require high resolution instrumentation. This becomes more challenging as sequence length and post-translational modification complexity increases. In this thesis, I will discuss my efforts in improving MS-based neuropeptidomic workflows, specifically in relation to "top-down" protein and peptide characterization, while highlighting improvements in identification and quantification of peptides through optimized fragmentation and cysteine reactive labeling. I will also discuss my contributions to our group's novel software, HyPep, enabling utility for confident neuropeptide identification. Altogether, these methods have contributed to increased neuropeptide coverage and can be translated to the analysis of other peptide classes.This thesis begins with two reviews of recent developments in MS-based analysis of neuropeptides. Each review outlines examples of how advancements in MS sample preparation, MS acquisition, data analysis, and technology has increased our understanding of neuropeptides, and provides insight on promising directions for further development within each section. My research contributions in Professor Lingjun Li's research group started by applying peptidomics workflows to quantify changes in neuropeptide expression due to hypoxia stress in a crustacean animal model, C. sapidus. Using the same animal model, we also increased the number of detectable neuropeptides using matrix-assisted laser desorption/ionization (MALDI)-MS imaging by incorporating novel tissue wash steps during sample preparation. Over 100 putative peptidomic biomarkers for different levels of hypoxia severity and duration were discovered within multiple tissue types. Four neuropeptide biomarkers were examined for functional activity and two neuropeptides were discovered to increase cardiac output ex vivo, including the first-ever reported function for a crustacean hyperglycemic hormone precursor-related peptide (CPRP). Since CPRP (3.5 kDa) is co-released with crustacean hyperglycemic hormone (CHH) neuropeptide (8.5 kDa), an important neuropeptide that regulates glucose metabolism, we did not detect CHH in these hypoxia studies because its larger size prevented confident detection using typical neuropeptidomic MS workflow and data acquisition methods. Previous members of our research group have developed MS methods to sequence CHH, but limitations of those methods included requiring multiple MS instruments or reliance on targeted MS strategies in which information of potential novel neuropeptides is lost. I built upon those MS acquisition and data analysis methods and was able to sequence novel CHH isoforms and proteoforms, without assistance from genomic or transcriptomic-based sequence prediction, using a Thermo Orbitrap Fusion Lumos instrument. Additionally, I developed novel sample preparation methods to reduce the disulfide bonds and alkylate all six cysteine residues on CHH at high efficiency (83-89% labeling efficiency). While developing methods for CHH are beneficial for analyzing mid-size (8-10 kDa) peptides, I sought to also advance general neuropeptidomics data analysis pipeline. A novel concept for neuropeptide identification software, called HyPep, was created 10 years ago but was not finished and could not be used. To make the software usable, I incorporated features typically used in peptide-spectrum matching (PSM)-based algorithms in proteomics software, such as a false discovery rate. I optimized the software parameters which resulted in HyPep outperforming an analogous commercial software in identifying neuropeptides from four crustacean neuronal tissue types. Collectively, this dissertation is devoted to advancing sample preparation, data acquisition, and data analysis strategies for neuropeptidomics, and these methods are also potentially translatable to analyze other classes of endogenous peptides.
Download or read book Mass Spectral Analysis of Crustacean Signaling Peptides Using a Multi dimensional Strategy written by and published by . This book was released on 2013 with total page 331 pages. Available in PDF, EPUB and Kindle. Book excerpt: By definition, neuropeptides are a class of signaling molecules either secreted by or that have an effect on neurons. They are present at minute concentrations in complex biological matrices. Their expression levels vary both temporally and spatially as they regulate several biological functions. Our limited knowledge of their regulatory processes has resulted in the creation or understanding of several therapeutic treatments with more advancements hinging on future neuropeptidomic breakthroughs. A multi-dimensional strategy has been implemented for the analysis of neuropeptides in the crustacean model system. Due to their dynamic nature, biological mass spectrometry is the instrumental platform used for neuropeptide analysis. It is coupled with analytical methodologies primarily focused on improving neuropeptide detection, identification, and quantitation. Simple protocols for rapid, robust, and reproducible analysis have been developed by modifying previous techniques for novel application on newer technologies. Several protocols were developed by acknowledging both the benefits and limitations of the techniques implemented. While this work utilizes both soft ionization techniques, more emphasis is given to the development of MALDI analysis. Relative quantitative analysis by 4-plex N,N-dimethyl leucine (DiLeu) labels was applied to study the neuropeptide expression level changes in relation to feeding. A neuropeptide database was compiled for automated analysis of large datasets using popular protein identification and quantitation software. Improving mass spectrometry de novo sequencing was achieved by modifying a simple acid-driven deamidation protocol to determine the presence and location of amides. Also, the formation of multiply charged peptides by MALDI ionization was explored to improve fragmentation spectra. Finally, to further analysis by MALDI ionization, proof-of-principle projects were initiated. The first was established to improve detection of hemolymph samples on newer MALDI instruments to determine expression level changes of circulating neuropeptides in relation to the circadian rhythm cycle. The second was the application of isotopic DiLeu (iDiLeu) labels to generate a five point standard curve for absolute quantitation. Primary interests of these labels are in their application for imaging experiments. In conclusion, all projects were developed by implementing a multi-dimensional strategy to strengthen the mass spectrometric platform for advancing neuropeptidomic research.
Download or read book Peptidomics written by Mikhail Soloviev and published by John Wiley & Sons. This book was released on 2007-12-21 with total page 432 pages. Available in PDF, EPUB and Kindle. Book excerpt: The definitive guide to peptidomics- a hands-on lab reference The first truly comprehensive book about peptidomics for protein and peptide analysis, this reference provides a detailed description of the hows and whys of peptidomics and how the techniques have evolved. With chapters contributed by leading experts, it covers naturally occurring peptides, peptidomics methods and new developments, and the peptidomics approach to biomarker discovery. Explaining both the principles and the applications, Peptidomics: Methods and Applications: * Features examples of applications in diverse fields, including pharmaceutical science, toxicity biomarkers, and neuroscience * Details the successful peptidomic analyses of biological material ranging from plants to mammals * Describes a cross section of analytical techniques, including traditional methodologies, emerging trends, and new techniques for high throughput approaches An enlightening reference for experienced professionals, this book is sufficiently detailed to serve as a step-by-step guide for beginning researchers and an excellent resource for students taking biotechnology and proteomics courses. It is an invaluable reference for protein chemists and biochemists, professionals and researchers in drug and biopharmaceutical development, analytical and bioanalytical chemists, toxicologists, and others.
Download or read book Fundamentals and Applications of Fourier Transform Mass Spectrometry written by Philippe Schmitt-Kopplin and published by Elsevier. This book was released on 2019-08-11 with total page 778 pages. Available in PDF, EPUB and Kindle. Book excerpt: Fundamentals and Applications of Fourier Transform Mass Spectrometry is the first book to delve into the underlying principles on the topic and their linkage to industrial applications. Drs. Schmitt-Kopplin and Kanawati have brought together a team of leading experts in their respective fields to present this technique from many different perspectives, describing, at length, the pros and cons of FT-ICR and Orbitrap. Numerous examples help researchers decide which instruments to use for their particular scientific problem and which data analysis methods should be applied to get the most out of their data. Covers FT-ICR-MS and Orbitrap’s fundamentals, enhancing researcher knowledge Includes details on ion sources, data processing, chemical analysis and imaging Provides examples across the wide spectrum of applications, including omics, environmental, chemical, pharmaceutical and food analysis
Download or read book MALDI Mass Spectrometry Imaging written by Tiffany Porta Siegel and published by Royal Society of Chemistry. This book was released on 2021-11-23 with total page 403 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book gathers, in a single resource, knowledge about matrix-assisted laser desorption ionisation (MALDI) mass spectrometry imaging. It includes fundamentals in the MALDI ionisation process, different source geometries and capabilities, detection systems and the latest research and applications in the range of –omics area as well as other broader areas. Chapters will touch on dedicated sample preparation protocols specific for the class of compounds of interest, instrumentation used with strengths and current limitations, strategies for structural analysis and identification and applications. It will be a welcomed addition to the literature in this fast-moving field and provide a guide to new innovations and applications especially in metabolomics and proteomics. With contributions from leading experts, this book will be an authoritative guide to this method. Aimed at postgraduate and professional researchers, in academia and in the industrial market where it has direct application to clinical research. It will be a supporting volume for those just entering the field as well as experienced practitioners.
Download or read book Neuroproteomics written by Oscar Alzate and published by CRC Press. This book was released on 2009-10-26 with total page 356 pages. Available in PDF, EPUB and Kindle. Book excerpt: In this, the post-genomic age, our knowledge of biological systems continues to expand and progress. As the research becomes more focused, so too does the data. Genomic research progresses to proteomics and brings us to a deeper understanding of the behavior and function of protein clusters. And now proteomics gives way to neuroproteomics as we beg
Download or read book Mass Spectrometry and Hyphenated Techniques in Neuropeptide Research written by Jerzy Silberring and published by John Wiley & Sons. This book was released on 2002-05-02 with total page 576 pages. Available in PDF, EPUB and Kindle. Book excerpt: The first authoritative guide to the application of this vital analytical technique Mass spectrometry is a powerful analytical technique that is used to identify unknown compounds, to quantify known materials, and to elucidate the structural and chemical properties of molecules. In analyzing the effects of experimental drugs on the brain, it is the sole technique for identifying the presence and structure of neuropeptides-substances that indicate the effect of the drug. Mass Spectrometry and Hyphenated Techniques in Neuropeptide Research specifically explains how to apply the technology to this process. Because the book is written by mass spectrometry users, as opposed to mass spectrometrists, the focus remains on practical applications of the technique. The authors demonstrate how mass spectrometry works, how to apply the technique to research, which types of instrumentation should be used for particular requirements, and how to plan experiments. Readers will learn why mass spectrometry provides more outcome features than other techniques in neuropeptide analysis, including simultaneous detection, identification of substances present in mixtures, and sequence information even when the residues are modified, blocked, or unusual. Among the chapters in this comprehensive text are: * Sequencing of Peptides by Nanospray Mass Spectrometry * Laser-Machined Microdevices for Mass Spectrometry * Electron Capture Dissociation of Peptides * Synthesis of Combinatorial Peptide Libraries * Analysis of Tissues That Reflect Nervous System Disease Doctoral students, researchers, and industry professionals in pharmacology, chemistry, biochemistry/biotechnology, and medicine will find Mass Spectrometry and Hyphenated Techniques in Neuropeptide Research to be an indispensable starting point for understanding peptides, their function, and identification.
Download or read book Proteomics for Biological Discovery written by Timothy D. Veenstra and published by John Wiley & Sons. This book was released on 2006-06-12 with total page 361 pages. Available in PDF, EPUB and Kindle. Book excerpt: Written by recognized experts in the study of proteins, Proteomics for Biological Discovery begins by discussing the emergence of proteomics from genome sequencing projects and a summary of potential answers to be gained from proteome-level research. The tools of proteomics, from conventional to novel techniques, are then dealt with in terms of underlying concepts, limitations and future directions. An invaluable source of information, this title also provides a thorough overview of the current developments in post-translational modification studies, structural proteomics, biochemical proteomics, microfabrication, applied proteomics, and bioinformatics relevant to proteomics. Presents a comprehensive and coherent review of the major issues faced in terms of technology development, bioinformatics, strategic approaches, and applications Chapters offer a rigorous overview with summary of limitations, emerging approaches, questions, and realistic future industry and basic science applications Discusses higher level integrative aspects, including technical challenges and applications for drug discovery Accessible to the novice while providing experienced investigators essential information Proteomics for Biological Discovery is an essential resource for students, postdoctoral fellows, and researchers across all fields of biomedical research, including biochemistry, protein chemistry, molecular genetics, cell/developmental biology, and bioinformatics.
Download or read book Behavioral Genetics of the Fly Drosophila Melanogaster written by Josh Dubnau and published by Cambridge University Press. This book was released on 2014-06-26 with total page 309 pages. Available in PDF, EPUB and Kindle. Book excerpt: A comprehensive portrayal of the behaviour genetics of the fruit fly (Drosophila melanogaster) and the methods used in these studies.
Download or read book Neuropeptide Systems as Targets for Parasite and Pest Control written by Timothy G. Geary and published by Springer Science & Business Media. This book was released on 2011-01-13 with total page 246 pages. Available in PDF, EPUB and Kindle. Book excerpt: The need to continually discover new agents for the control or treatment of invertebrate pests and pathogens is undeniable. Agriculture, both animal and plant, succeeds only to the extent that arthropod and helminth consumers, vectors and pathogens can be kept at bay. Humans and their companion animals are also plagued by invertebrate parasites. The deployment of chemical agents for these purposes inevitably elicits the selection of resistant populations of the targets of control, necessitating a regular introduction of new kinds of molecules. Experience in other areas of chemotherapy has shown that a thorough understanding of the biology of disease is an essential platform upon which to build a discovery program. Unfortunately, investment of research resources into understanding the basic physiology of invertebrates as a strategy to illuminate new molecular targets for pesticide and parasiticide discovery has been scarce, and the pace of introduction of new molecules for these indications has been slowed as a result. An exciting and so far unexploited area to explore in this regard is invertebrate neuropeptide physiology. This book was assembled to focus attention on this promising field by compiling a comprehensive review of recent research on neuropeptides in arthropods and helminths, with contributions from many of the leading laboratories working on these systems.
Download or read book Chemical Communication in Crustaceans written by Thomas Breithaupt and published by Springer Science & Business Media. This book was released on 2010-11-25 with total page 572 pages. Available in PDF, EPUB and Kindle. Book excerpt: The crustaceans are ecologically and economically important organisms. They constitute one of the dominant invertebrate groups on earth, particularly within the aquatic realm. Crustaceans include some of the preferred scientific model organism, profitable aquaculture specimen, but also invasive nuisance species threatening native animal communities throughout the world. Chemoreception is the most important sensory modality of crustaceans, acquiring important information about their environment and picking up the chemical signals that mediate communication with conspecifics. Significant advances have been made in our understanding of crustacean chemical communication during the past decade. This includes knowledge about the identity, production, transfer, reception and behavioral function of chemical signals in selected crustacean groups. While it is well known that chemical communication is an integral part of the behavioral ecology of most living organisms, the intricate ways in which organisms allocate chemicals in communication remains enigmatic. How does the environment influence the evolution of chemical communication? What are the environmental cues that induce production or release of chemicals? How do individuals economize production and utilization of chemicals? What is the importance of molecule specificity or mix of a molecule cocktail in chemical communication? What is the role of chemical cues in multimodal communication? How does the ontogenetic stage, the sex or the physiological status of an individual affect its reaction to chemical cues? Many of these questions still represent important challenges to biologists.
Download or read book Dynamic Biological Networks written by Ronald M. Harris-Warrick and published by MIT Press. This book was released on 1992 with total page 366 pages. Available in PDF, EPUB and Kindle. Book excerpt: This introduction to the crustacean stomatogastric nervous system (STNS) describes some of the best-understood neural networks in the animal kingdom at cellular, network, behavioural, comparative and evolutionary levels of analysis.
Download or read book The Crustacean Nervous System written by Konrad Wiese and published by Springer Science & Business Media. This book was released on 2002 with total page 652 pages. Available in PDF, EPUB and Kindle. Book excerpt: This unique selection of reviews summarizes current knowledge in all major fields of crustacean neurobiology and all levels of their CNS organization, using lobster and crayfish. It not only imparts theoretical knowledge but also describes all available contemporary and advanced techniques, such as patch clamp recordings, microelectrode techniques, immunocytochemistry, and all methods of molecular genetics to identify cellular pathways of protein synthesis and peptidergic control. In summary, it is a comprehensive account of the research achievements in one of the major nervous systems besides the mammalian CNS.
Download or read book Protein and Peptide Mass Spectrometry in Drug Discovery written by Michael L. Gross and published by John Wiley & Sons. This book was released on 2011-09-26 with total page 484 pages. Available in PDF, EPUB and Kindle. Book excerpt: The book that highlights mass spectrometry and its application in characterizing proteins and peptides in drug discovery An instrumental analytical method for quantifying the mass and characterization of various samples from small molecules to large proteins, mass spectrometry (MS) has become one of the most widely used techniques for studying proteins and peptides over the last decade. Bringing together the work of experts in academia and industry, Protein and Peptide Mass Spectrometry in Drug Discovery highlights current analytical approaches, industry practices, and modern strategies for the characterization of both peptides and proteins in drug discovery. Illustrating the critical role MS technology plays in characterizing target proteins and protein products, the methods used, ion mobility, and the use of microwave radiation to speed proteolysis, the book also covers important emerging applications for neuroproteomics and antigenic peptides. Placing an emphasis on the pharmaceutical industry, the book stresses practice and applications, presenting real-world examples covering the most recent advances in mass spectrometry, and providing an invaluable resource for pharmaceutical scientists in industry and academia, analytical and bioanalytical chemists, and researchers in protein science and proteomics.
