Main description:

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.

Contents:

Preface...
Table of Contents...
Contributing Authors...

Part I Methods for the Detection of Melatonin and Its Biological Derivatives

1. Two-Dimensional Thin Layer Chromatography of Melatonin and Related Compounds
David C. Klein

2. Detection of Melatonin in Tissue Samples
Celine Legros and Jean A. Boutin

3. Measuring Dim Light Melatonin Onset in Humans
Melissa A. St. Hilaire and Steven W. Lockley

4. Measuring Urinary 6-sulphatoxymelatonin in Humans
Melissa A. St. Hilaire and Steven W. Lockley

5. MEL-Index: Estimation of Tissue Melatonin Levels using Gene Expression Data
Pedro Augusto CM Fernandes, Regina P. Markus, and Gabriela Sarti Kinker

6. Melatonin Synthesis Enzymes Activity: Radiometric Assays for AANAT, ASMT, and TPH
Fernanda Gaspar do Amaral, Jose Cipolla-Neto, and Solange Castro Afeche

7. Pinealectomy in Rats
Camila Congentino Gallo, Fernanda Akane Nishino, Fernanda Gaspar do Amaral, and Jose Cipolla-Neto

8. Surgical Techniques and Nuances for Superior Cervical Ganglionectomy and Decentralization in Rats
Sarosh Irfan Madhani, David C. Klein, Estela M. Munoz, and Luis E. Savastano

9. Pineal Microdialysis
Caroline Aparecida Pereira de Souza, Fernanda Akane Nishino, Fernanda Gaspar do Amaral, and Jose Cipolla-Neto

10. Radiochemical In Situ Hybridization in Developmental Studies of the Pineal Gland
Martin Fredensborg Rath and Morten Moller

11. Pineal Cells Dissociation and Culture: Isolated Pinealocytes, Isolated Astrocytes, and Co-Culture
Solange Castro Afeche, Diego de Piazza Pimentel, Luis Felipe Ferro, and Jose Cipolla-Neto

12. Pineal Gland Culture
Solange Castro Afeche, Fernanda Gaspar do Amaral, and Jose Cipolla-Neto

13. Preparation of Single Pineal Cells
Steven L. Coon and David C. Klein

14. RNA Sequencing of Single Pineal CellsSteven L. Coon and David C. Klein

15. siRNA-Mediated Downregulation of Gene Expression in Cultured Rat Pineal Cells
Aurea Susana Blancas-Velazquez and Martin Fredensborg Rath

16. Analysis of the Human Pineal Proteome by Mass Spectrometry
Mariette Matondo, Guillaume Dumas, and Erik Maronde

17. The Pineal Transcriptome Webpage
David C. Klein, Steven L. Coon, Carol Ensault, and Dale Ryan

Part II Methods to Measure Melatonin Effects at the Molecular and Cellular Level

18. 2-[125I]iodomelatonin and [3H]melatonin Binding Assays for Melatonin Receptors
Florence Gbahou and Ralf Jockers

19. Alternative Ligands at Melatonin Receptors
Celine Legros, Said Yous, and Jean A. Boutin

20. GTP S Binding Assay for Melatonin Receptors in Mouse Brain Tissue
Romain Gerbier and Ralf Jockers

21. MT1 Melatonin Receptor Reconstitution in Nanodiscs
Jean A. Boutin, Christel Logez, Marjorie Damian, Renaud Wagner, Jean-Louis Baneres, and Gilles Ferry

22. Functional Investigation of Melatonin Receptors Activation by Homogenous cAMP Assay
Erika Cecon, Jean-Luc Guillaume, and Ralf Jockers

23. Functionality of Melatonin Receptors: Internalization
Clemence Dupre, Celine Legros, and Jean A. Boutin

24. Functionality of Melatonin Receptors: Recruitment of -arrestin at MT1
Clemence Dupre, Celine Legros, and Jean A. Boutin

25. MT1 Receptor Signaling Pathways by Impedance Measurement
Anne Bonnaud, Clemence Dupre, Celine Legros, and Jean A. Boutin

26. Measuring Protein-Protein Interactions of Melatonin Receptors by Bioluminescence Resonance Energy Transfer (BRET)
Atsuro Oishi and Ralf Jockers

27. Non-Radioactive In Situ Hybridization of MT1 Melatonin Receptor for the Identification of Melatonin Target Cells
Paul Klosen

28. Beta-Galactosidase as a Transgenic Reporter for the Mapping and Phenotyping of MT1 and MT2 Melatonin Receptor Expressing Cells
Paul Klosen

29. Why Are We Still Cloning Melatonin Receptors? A Commentary
Celia Gautier, Isabelle Theret, Giulia Lizzo, Gilles Ferry, Sophie-Penelope Guenin, and Jean A. Boutin

30. Measuring Binding at the Putative Melatonin Receptor MT3
Celine Legros, Philippe Dupuis, Gilles Ferry, and Jean A. Boutin

31. Cloning, Expression, Purification, Crystallization, and X-Ray Structural Determination of the Human NQO2 in Complex with Melatonin
Barbara Calamini, Gilles Ferry, and Jean A. Boutin

32. Melatonin Binding to Human NQO2 by Isothermal Titration Calorimetry
Barbara Calamini, Gilles Ferry, and Jean A. Boutin

33. Measurement of NQO2 Catalytic Activity and of Its Inhibition by Melatonin
Gilles Ferry and Jean A. Boutin

34. Measuring the NQO2: Melatonin Complex by Native Nano-Electrospray Ionization Mass Spectrometry
Jean A. Boutin, Johann Stojko, Gilles Ferry, and Sarah Cianferani

35. Human and Rodent Cell Lines as Models of Functional Melatonin-Responsive Pancreatic Islet Cells
Juliane Zibolka, Ina Bahr, Elmar Peschke, Eckhard Muhlbauer, and Ivonne Bazwinsky-Wutschke

36. Mesenchymal Stem Cell and Monocyte Co-Cultures
Fahima Munmun and Paula A. Witt-Enderby

37. Real-Time Monitoring of Circadian Rhythms in the Eye
Kenkichi Baba and Gianluca Tosini

Part III Methods to Measure Melatonin Effects In Vivo

38. Assessing the Role of Melatonin in the Modulation of Visual Functions in the Mouse
Kenkichi Baba and Gianluca Tosini

39. Methods to Assess Melatonin-Receptor Mediated Phase-Shift and Re-Entrainment of Rhythmic Behaviors in Mouse Models
Grant C. Glatfelter, Jennifer Sosa, Randall L. Hudson, and Margarita L. Dubocovich

40. Neurotrasmission Recovery by Melatonin Measured by CMAP
Samuele Negro, Marco Stazi, Michela Rigoni, and Aram Megighian

41. Olfaction and Melatonin: The Use of the Olfactory Discrimination Test
Ana Carolina Duarte Noseda and Marcelo M. S. Lima

42. Photothrombotic Mouse Models for the Study of Melatonin as a Therapeutic Tool After Ischemic Stroke
Marco Cambiaghi, Laura Cherchi, and Stefano Comai

43. Hypnotic Effects of Melatonergic Compounds Measured in Mice or Rats
Stefano Comai and Danilo De Gregorio

44. Acute and Chronic Pain Preclinical Models to Study the Analgesic Properties of Melatonergic Compounds
Danilo De Gregorio and Stefano Comai

45. Behavioral and Fluorescent-Based Immunohistochemistry Protocols for Examining Anti-Depressant-Like Effects of Melatonin in Mice
Estrada-Reyes Rosa, Munoz Estrada Jesus, and Benitez-King Gloria

46. A Method for Perfusion of Tissue-Isolated Human Tumor Xenografts in Nude Rats to Investigate the Oncostatic Role of the Physiological Nocturnal Melatonin Signal
Robert T. Dauchy, Steven M. Hill, and David E. Blask

47. A Method for Growing Tissue-Isolated Human Tumor Xenografts in Nude Rats for Melatonin/Cancer Studies
Robert T. Dauchy, Steven M. Hill, and David E. Blask

Subject Index List...