Calcium imaging of gut activity.

摘要

The major cell types regulating gut motility include enteric neurones, interstitial cells of Cajal (ICC) and their effector smooth muscle cells. These cells are arranged conveniently in nested layers through the gut wall. Our knowledge of how many of these cells in each layer are integrated to produce the various patterns of motility is largely unknown. So far, much of our knowledge of gut motility has usually been obtained by examining point sources of activity (e.g. intracellular recordings from enteric neurones, ICC and smooth muscle cells), rather than the spread of activity through these spatially distributed nerve and ICC networks, or smooth muscle syncitia. Our understanding of how these cells are integrated to produce gut movements would be greatly enhanced if we could image the activity in many of these cells in each layer, or many cells in several layers, simultaneously. Calcium (Ca2+) is a major signalling and regulatory molecule in most cells. In fact, electrical excitability in enteric neurones, ICC and smooth muscle is associated with robust rises in intracellular Ca2+ that long outlast the electrical events (e.g. action potentials in neurones and smooth muscle) that gave rise to them. These prolonged Ca2+ responses, together with the development of several high quality Ca2+ indicators, has provided a unique opportunity to image many cells in intact tissues simultaneously using ICCD video-rate cameras along with conventional microscopy. However, confocal microscopy has also been used, and has several advantages over the above systems. These include reduced photo-toxicity and bleaching and the elimination of out of focus light from different layers within the tissue. So far, despite some limitations with the calcium imaging techniques, the spread of activity through the two layers of smooth muscle, ICC networks and myenteric neurones in intact preparations, or cultured myenteric neuronal networks, is beginning to yield exciting new data about how these different cells interact and process information.