图书简介
This volume covers the latest techniques that study the synthesis of melatonin, its receptor function, and its effects at the cellular and systemic level. The chapters are organized into three parts. Part One describes methods for the detection of melatonin and its biological derivatives in various biological samples, the manipulation of melatonin synthesis by the pineal gland in animals, and the principal source of melatonin in mammals. Part Two explores methods to measure the biological effects and consequences of melatonin binding to high-affinity G protein-coupled receptors. Part Three describes methods to measure the physiological effects that are regulated by melatonin in animals, particularly in rodent models. 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, Melatonin: Methods and Protocols is a valuable resource for any researcher interested in investigating melatonin, from its production to its mechanisms of action and systemic effects.
Two-Dimensional Thin Layer Chromatography of Melatonin and Related Compounds.- Detection of Melatonin in Tissue Samples.- Measuring Dim Light Melatonin Onset in Humans.- Measuring Urinary 6-sulphatoxymelatonin in Humans.- MEL-Index: Estimation of Tissue Melatonin Levels using Gene Expression Data.- Melatonin Synthesis Enzymes Activity: Radiometric Assays for AANAT, ASMT, and TPH.- Pinealectomy in Rats.- Surgical Techniques and Nuances for Superior Cervical Ganglionectomy and Decentralization in Rats.- Pineal Microdialysis.- Radiochemical In Situ Hybridization in Developmental Studies of the Pineal Gland.- Pineal Cells Dissociation and Culture: Isolated Pinealocytes, Isolated Astrocytes, and Co-Culture.- Pineal Gland Culture.- Preparation of Single Pineal Cells.- RNA Sequencing of Single Pineal Cells.- siRNA-Mediated Downregulation of Gene Expression in Cultured Rat Pineal Cells.- Analysis of the Human Pineal Proteome by Mass Spectrometry.- The Pineal Transcriptome Webpage.- 2-[125I]iodomelatonin and [3H]melatonin Binding Assays for Melatonin Receptors.- Alternative Ligands at Melatonin Receptors.- GTPS Binding Assay for Melatonin Receptors in Mouse Brain Tissue.- MT1 Melatonin Receptor Reconstitution in Nanodiscs.- Functional Investigation of Melatonin Receptors Activation by Homogenous cAMP Assay.- Functionality of Melatonin Receptors: Internalization.- Functionality of Melatonin Receptors: Recruitment of β-arrestin at MT1.- MT1 Receptor Signaling Pathways by Impedance Measurement.- Measuring Protein-Protein Interactions of Melatonin Receptors by Bioluminescence Resonance Energy Transfer (BRET).- Non-Radioactive In Situ Hybridization of MT1 Melatonin Receptor for the Identification of Melatonin Target Cells.- Beta-Galactosidase as a Transgenic Reporter for the Mapping and Phenotyping of MT1 and MT2 Melatonin Receptor Expressing Cells.- Why Are We Still Cloning Melatonin Receptors? A Commentary.- Measuring Binding at the Putative Melatonin Receptor MT3.- Cloning, Expression, Purification, Crystallization, and X-Ray Structural Determination of the Human NQO2 in Complex with Melatonin.- Melatonin Binding to Human NQO2 by Isothermal Titration Calorimetry.- Measurement of NQO2 Catalytic Activity and of Its Inhibition by Melatonin.- Measuring the NQO2: Melatonin Complex by Native Nano-Electrospray Ionization Mass Spectrometry.- Human and Rodent Cell Lines as Models of Functional Melatonin-Responsive Pancreatic Islet Cells.- Mesenchymal Stem Cell and Monocyte Co-Cultures.- Real-Time Monitoring of Circadian Rhythms in the Eye.- Assessing the Role of Melatonin in the Modulation of Visual Functions in the Mouse.- Methods to Assess Melatonin-Receptor Mediated Phase-Shift and Re-Entrainment of Rhythmic Behaviors in Mouse Models.- Neurotrasmission Recovery by Melatonin Measured by CMAP.- Olfaction and Melatonin: The Use of the Olfactory Discrimination Test.- Photothrombotic Mouse Models for the Study of Melatonin as a Therapeutic Tool After Ischemic Stroke.- Hypnotic Effects of Melatonergic Compounds Measured in Mice or Rats.- Acute and Chronic Pain Preclinical Models to Study the Analgesic Properties of Melatonergic Compounds.- Behavioral and Fluorescent-Based Immunohistochemistry Protocols for Examining Anti-Depressant-Like Effects of Melatonin in Mice.- A Method for Perfusion of Tissue-Isolated Human Tumor Xenografts in Nude Rats to Investigate the Oncostatic Role of the Physiological Nocturnal Melatonin Signal.- A Method for Growing Tissue-Isolated Human Tumor Xenografts in Nude Rats for Melatonin/Cancer Studies.
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