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A complete understanding of how behavior is controlled by the nervous system must include detailed knowledge of the underlying circuitry and neurochemistry. The parasitic nematode Ascaris suum, with only 298 neurons, is an exceptionally well-suited system in which to obtain this level of detail. Whereas a great deal of the cytoarchitecture, electrophysiological properties, and classical transmitter phenotypes of individual cells has been described, much is still unknown about the great diversity of modulatory neuropeptides. A description of their structure and activity is essential for understanding the in the control of behavior. In this thesis, we use a combination of mass spectrometry (MS), bioinformatics, and molecular biology to discover and localize neuropeptides in the motor nervous system. The peptides are then chemically synthesized and tested for bioactivity. Direct profiling of single neurons by matrix assisted laser desorption/ ionization time-of-flight (MALDI-TOF) MS identified diverse and interesting patterns of peptide expression. In parts one and two, we targeted ventral cord motorneurons that innervate the somatic body wall musculature responsible for locomotion. In the cholinergic excitatory motorneurons, two different peptide expression profiles were observed. The first was found in neurons responsible for forward locomotion, and the second, a completely different set of peptides, was present in neurons responsible for backwards locomotion. Aside from confirming the identity of previously known peptides, all of which had been shown to be bioactive, we also identified six novel peptides. In the second part, in the GABAergic inhibitory motorneurons we found a novel bioactive peptide with potent inhibitory effects on muscle contraction. In the third part, we mapped peptide expression in the 4 GABAergic RME ring motorneurons neurons of the head. The peptides in these neurons, of which 7 were novel, were different from those in the ventral cord inhibitors. Three peptides encoded by one of the transcripts had inhibitory effects on muscle contraction. This is the first study to identify peptide expression patterns in the motor neurons. In this study we have not only found novel peptides but also identified the peptides that are likely to be most relevant to locomotion in A. suum.
Analysis of Neuropeptides by Liquid Chromatography and Mass Spectrometry
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.
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.
This book tackles a number of different perspectives concerning the parasitic helminth Ascaris, both in animals and in humans and the disease known as ascariasis. It seeks to identify interesting, exciting and novel aspects, which will interest readers from a broad range of disciplines.Over a quarter of the world's population are infected with the human roundworm, and the equivalent in pigs is equally ubiquitous. Both contribute to insidious and chronic nutritional morbidity, and this has been quantified, in humans, as disability adjusted life years approximating 10.5 million. Ascaris larvae develop in host parenteral tissues, and the resultant pathology has been condemnation. Ascariasis, despite its staggering global prevalence and the sheer numbers of people it infects, remains a classic neglected disease. However, renewed interest in the consequences of early infection with worms from the perspective of immune modulation, co-infections and the development of allergy further enhances the relevance of these parasites. - Brings together a wide range of topics and approaches and recent, comprehensive and progressive research concerning the neglected parasite Ascaris - Provides a blueprint of how a single parasite entity can stimulate interest in basic biology, clinical science, veterinary science, public health and epidemiology - Presents a wealth of new insights given that a book on this parasite has not been published for over 20 years - 16 chapters from a range of top authors from around the world
The structural simplicity of nematode nervous systems has provided a useful resource for studies on basic neurobiology. Two species have underpinned basic neuroscience research in phylum Nematoda, the free-living nematode Caenorhabditis elegans and the pig parasitic nematode Ascaris suum due to their genetic tractability and large size, respectively. The fact that most of the chemotherapeutic targets for the treatment of roundworm parasite infections of animals and man associate with neuromuscular function means that nematode neurobiology research resonates beyond discovery biology and informs parasite control. These diverse drivers for the interrogation of nematode neurobiology have exposed a unique approach to the regulation of complex behavioral traits, encompassing structural simplicity with profound chemical complexity. The latter embraces extreme diversity in both the ligand-gated ion channel subunit complements for classical neurotransmitters and in the neuropeptide ligands that signal mostly via G-protein coupled receptors, offering compelling opportunities for parasite control-target discovery and validation.