Melatonin: Methods and Protocols

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
C幨ine 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, Jos?Cipolla-Neto, and Solange Castro Afeche

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

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

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

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

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

12. Pineal Gland Culture
Solange Castro Afeche, Fernanda Gaspar do Amaral, and Jos?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
C幨ine 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 Ban鋨es, 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
Cl幦ence Dupr? C幨ine Legros, and Jean A. Boutin

24. Functionality of Melatonin Receptors: Recruitment of β-arrestin at MT1
Cl幦ence Dupr? C幨ine Legros, and Jean A. Boutin

25. MT1 Receptor Signaling Pathways by Impedance Measurement
Anne Bonnaud, Cl幦ence Dupr? C幨ine 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 Mel