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.
Inhaltsverzeichnis
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.
