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Free cytosolic Ca 2+ recordings from myenteric neurones in multilayer intestinal preparations[Note 1. Parts of this study have been published in abstract ...]
Author(s) -
Berghe P. Vanden,
Missiaen L.,
Bellon E.,
Vanderwinden JM.,
Janssens J.,
Tack J.
Publication year - 2001
Publication title -
neurogastroenterology and motility
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.489
H-Index - 105
eISSN - 1365-2982
pISSN - 1350-1925
DOI - 10.1046/j.1365-2982.2001.00283.x
Subject(s) - myenteric plexus , depolarization , confocal , enteric nervous system , electrophysiology , confocal microscopy , acetylcholine , chemistry , interstitial cell of cajal , differential interference contrast microscopy , ganglion , neuroscience , biophysics , anatomy , biology , endocrinology , medicine , microbiology and biotechnology , pathology , microscopy , immunohistochemistry , smooth muscle , geometry , mathematics
The ability to simultaneously monitor different myenteric neurones in a multilayer preparation may enhance our understanding of the enteric nervous system. Longitudinal muscle myenteric plexus prepar‐ ations were mounted in recording chambers with a coverslip base and loaded with Indo‐1‐AM. cytosolic Ca 2+ concentration ([Ca 2+ ] i ); changes were recorded at room temperature with a confocal microscope. In addition to mechanical (pressure‐ring) and pharmacological (nifedipine) reduction of muscle contractions, purpose‐designed software was developed to reposition regions of interest and avoid artefacts. Confocal scanning permitted optical selection of single cell layers. High K + depolarization, used to distinguish between excitable and nonexcitable cells, caused a synchronous [Ca 2+ ] i rise in 84.3% of the ganglion cells. Acetylcholine, substance P and serotonin (all at 10 −5 mol L −1 ) induced transient [Ca 2+ ] i changes in subpopulations of myenteric neurones (45.1%, 42.9 and 21.9%, respectively). In addition to immediate responses to agonists, delayed [Ca 2+ ] i changes were also recorded, suggesting the presence of both directly activated and synaptically driven neurones. Functionally identified neurones and other cells in close apposition to the ganglia (interstitial cells of Cajal) could also be studied. This study demonstrates the potential of optical Ca 2+ recordings to monitor spread of activity in myenteric neurones and to study their interaction with non‐neuronal targets.