Use of Genetically Encoded Calcium Indicators (GECIs) Combined with Advanced Motion Tracking Techniques to Examine the Behavior of Neurons and Glia in the Enteric Nervous System of the Intact Murine Colon.

摘要: Genetically encoded Ca2+ indicators (GECIs) have been used extensively in many body systems to detect transients associated with neuronal activity. Their adoption enteric neurobiology has slower, although they offer advantages terms of selectivity, signal-to-noise and non-invasiveness. Our aims were utilize a number cell-specific promoters express the indicator GCaMP3 different classes neurons glia determine their effectiveness measuring activity neural networks during colonic motor behaviors. We developed several mice: 1) Wnt1-GCaMP3, all glia; 2) GFAP-GCaMP3, 3) nNOS-GaMP3, nitrergic neurons, 4) ChAT-GCaMP3, cholinergic neurons. These mice allowed us study behavior intact colon maintained at physiological temperature, especially migrating complex (CMMC), using low power imaging. In this preliminary study, we observed glial cell specific cells both myenteric submucous plexus transgenic variants. The that could be simultaneously imaged (100-1000 active cells) through undissected gut required advanced motion tracking analysis routines. pattern showed significant differences response spontaneous, oral or anal stimulation. Brief elongation mucosal stimulation, which evokes CMMC, most effective stimuli elicited powerful synchronized prolonged burst when compared same spontaneous CMMC. contrast, elongation, normally inhibits CMMCs, appeared suppress some an anally evoked follow but continued long after had waned. With these new tools unprecedented level detail can recorded from nervous system (ENS) minimal manipulation tissue. techniques extended order better understand roles particular normal disordered motility.