Study Purpose: GCaMP imaging combined with pharmacology is used as a surrogate for subthreshold and action potential activity to better understand the colonic neural circuit.
Data Collection: Mice expressing calcium indicator GCaMP6f were used to examine spontaneous and pharmacologically evoked changes in GCaMP-mediated fluorescence. Fluorescence was used as a surrogate for action potentials and neural circuit dynamics.
Primary Conclusion: None stated
Experimental Design: Excised colon segments were opened along the mesenteric border and pinned mucosal-side down onto a Sylgard surface lining a glass coverslip attached to the bottom of a plastic imaging chamber containing Krebs solution. Spontaneous and evoked changes in GCaMP6f-mediated Ca2+ fluorescence intensity were acquired in 12-bit images using a 1.44-megapixel CMOS camera capable of capturing at up to 80 frames/sec (Prime 95B; Teledyne Photometrics, Tuscon, AZ) controlled by MetaMorph software (version 188.8.131.52; Molecular Devices, Silicon Valley, CA). Image stacks of 400–1800 frames were processed, and motion-corrected when necessary using Fiji (National Institutes of Health, Bethesda, MD) software (version 2.0.0-rc-65/1.52a). DMPP (dimethylphenyl-piperazinium) was used to assay functional nAChRs (nicotinic acetylcholine receptors). The agonist was focally applied by pressure microperfusion (10 psi, 10 sec; via Picospritzer II; Parker Instrumentation Corp, Barnstaple, United Kingdon) from blunt glass micropipettes (diameter, 5–10 μm) delivered within 50 μm of an adjacent myenteric ganglion.
Completeness: This dataset is part of a larger study: "Quantitative Morphology, Transcriptomics, and Function of Enteric Neurons Expressing VIP"
Subjects & Samples: This study used one 120 day old male VIP-GCAmP reporter mouse.
Primary vs derivative data: Primary data contains imaging of calcium dynamics (GCaMP) in mouse colon as 1800 frames image stack .tif file. There is no derivative data folder.
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