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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.
List of contents
Rat pheochromocytoma PC12 cells in culture.- Adrenal grafts in the central nervous system: Chromaffin and chromaffin progenitor cell transplantation.- Immunocytochemistry of acutely isolated adrenal medullary chromaffin cells.- Transmission electron microscopy: a method for studying the adrenochromaffin cells.- Immunogold for protein location in chromaffin cells.- Visualization of exo- and endocytosis membrane dynamics with super-resolution STED microscopy.- Online detection of catecholamine release from the perfused rat adrenal gland.- On-line recording of perifused chromaffin cells.- Recording of chromaffin cell electrical activity in situ in acute adrenal slices.- Calcium imaging and amperometric recording in cultured chromaffin cells and adrenal slices from normotensive, Wistar Kyoto rats, and Spontaneously Hypertensive Rats.- Measurements of calcium in chromaffin cell organelles using targeted aequorins.- Quantification of secretory granule exocytosis by TIRF imaging and capacitance measurements.- Membrane capacitance measurements of stimulus-evoked exocytosis in adrenal chromaffin cell.- Quantal release analysis of electrochemically active molecules using single-cell amperometry.- Methodologies for detecting quantal exocytosis in adrenal chromaffin cells through diamond-based MEAs.- Vesicle collision protocols for the study of quantum size and exocytotic fraction released.- Patch Amperometry and Intracellular Patch Electrochemistry.- Artificial cells for dissecting exocytosis.- Isolation and purification of chromaffin granules from adrenal glands and cultured neuroendocrine cells.- Confocal microscopy studies of F-actin cytoskeleton distribution and dynamics using fluorescent LifeAct constructs in bovine adrenal chromaffin cells.- Unveiling the nanoscale dynamics of the exocytic machinery in chromaffin cells with single-molecule imaging.- Determinationof catecholamines in a small volume (25µl) of plasma from conscious mouse tail vein.- Quantification of chromogranin A and its fragments in biological fluids.- Cytotoxicity models in chromaffin cells to evaluate neuroprotective compounds.
