Main description:

This volume covers the most up-to-date methods and techniques used to further the understanding of chromaffin cell biology and pharmacology. Chapters guide readers through the basic mechanisms that regulate the stimulus-secretion coupling, chromaffin, tumor-derived cell PC-12 , morphology, biochemistry, pharmacology, electrophysiology, and electrochemistry. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Chromaffin Cells: Methods and Protocols aims to be a useful practical guide to researches to help further their study in this field.

Contents:

Part I Culturing and morphology

1. Rat pheochromocytoma PC12 cells in culture

Chin-Tien Wang

2. Adrenal grafts in the central nervous system: Chromaffin and chromaffin progenitor cell transplantation

Alejandra Boronat-Garcia, Marcela Palomero-Rivero, Magdalena Guerra-Crespo

3. Immunocytochemistry of acutely isolated adrenal medullary chromaffin cells

Keita Harada, Hidetada Matsuoka, Masumi Inoue

4. Transmission electron microscopy: a method for studying the adrenochromaffin cells

Anna Capaldo, Maria De Falco, Luigi Rosati and Vincenza Laforgia

5. Immunogold for protein location in chromaffin cells

Rafael Lujan, Rocio Alfaro-Ruiz and Carolina Aguado

6. Visualization of exo- and endocytosis membrane dynamics with super-resolution STED microscopy

Chung Yu Chan, Sue Han, Xin Wang, Xiaoli Guo and Ling-Gang Wu

Part II Stimulus-secretion coupling

7. Online detection of catecholamine release from the perfused rat adrenal gland

Ricardo Borges, Ana de la Iglesia and Antonio G. Garcia

8. On-line recording of perifused chromaffin cells

Ricardo De Pascual and Luis Gandia

9. Recording of chromaffin cell electrical activity in situ in acute adrenal slices

Nathali Guerineau

10. Calcium imaging and amperometric recording in cultured chromaffin cells and adrenal slices from normotensive, Wistar Kyoto rats, and Spontaneously Hypertensive Rats

Tzitzitlini Alejandre-Garcia, Pedro Segura-Chama, Oscar Javier Parada-Parra, Diana Millan-Aldaco and Arturo Hernandez-Cruz.

11. Measurements of calcium in chromaffin cell organelles using targeted aequorins.

Jaime Santo-Domingo, Pilar Alvarez-Illera, Pablo Montenegro, Rosalba I Fonteriz, Mayte Montero and Javier Alvarez

12. Quantification of secretory granule exocytosis by TIRF imaging and capacitance measurements

Muhmmad Omar-Hmeadi, Liangwen Liu, Santiago Echeverry, and Sebastian Barg

13. Membrane capacitance measurements of stimulus-evoked exocytosis in adrenal chromaffin cell

Elizabeth P. Seward and Robert C. Wykes

14. Quantal release analysis of electrochemically active molecules using single-cell amperometry

Natalia Dominguez, Pablo Montenegro and J. David Machado

15. Methodologies for detecting quantal exocytosis in adrenal chromaffin cells through diamond-based MEAs

Giulia Tomagra, Claudio Franchino, Emilio Carbone, Andrea Marcantoni, Alberto Pasquarelli, Federico Picollo, Valentina Carabelli

16. Vesicle collision protocols for the study of quantum size and exocytotic fraction released

Soodabeh Majdi, Alex S. Lima and Andrew Ewing

17. Patch Amperometry and Intracellular Patch Electrochemistry

Eugene V. Mosharov and Manfred Lindau

18. Artificial cells for dissecting exocytosis

Yuanmo Wang and Ann-Sofie Cans

Part III Chromaffin granules and membrane traffic

19. Isolation and purification of chromaffin granules from adrenal glands and cultured neuroendocrine cells

Arlek Gonzalez-Jamett, Maria Constanza Maldifassi, Ana Maria Cardenas

20. Confocal microscopy studies of F-actin cytoskeleton distribution and dynamics using fluorescent LifeAct constructs in bovine adrenal chromaffin cells

Jose Villanueva, Yolanda Gimenez-Molina and Luis M. Gutierrez

21. Unveiling the nanoscale dynamics of the exocytic machinery in chromaffin cells with single-molecule imaging
Adekunle T. Bademosi and Frederic A. Meunier

Part IV Miscellanea

22. Determination of catecholamines in a small volume (25l) of plasma from conscious mouse tail vein

Kechun Tang and Sushil K. Mahata

23. Quantification of chromogranin A and its fragments in biological fluids

Flavio Curnis, Barbara Colombo and Angelo Corti

24. Cytotoxicity models in chromaffin cells to evaluate neuroprotective compounds

Maria F. Cano Abad and Manuela G. Lopez