Activity of and neurochemical expression in nitrergic and cholinergic neurons in the murine colonic myenteric plexus

Study Purpose: Phasic calcium transient responses to different physiological stimuli, as well as neurochemical expression, in nitrergic and cholinergic myenteric neurons of the murine colon.

Data Collection: Activity of, and expression of neurochemicals within nitrergic and cholinergic myenteric neurons of the murine colon, using nNOS-CreERT2 and ChAT-Cre mice crossed to conditional GCaMP6f-expressing mice. Activity, in the form of Ca2+ transients, was imaged for two minutes at 10X in the middle colon during spontaneous colonic migrating motor complexes (CMMCs), in response to CMMCs caused by brush stimulation of the mucosa of the proximal or distal colon 30 seconds into the video (labeled in files as "oral" or "anal brush stim," respectively), in response to elongation of the proximal or distal colon 30 seconds into the video, or in response to direct electrical field stimulation (EFS) of the middle colon 30 seconds into the video.

Primary Conclusion: None stated.

Curator's Notes

Experimental Design: Adult mice of each sex were killed by isoflurane inhalation, and cervical dislocation, in accordance with the requirements of the Animal Ethics Committee at the University of Nevada, Reno. A ventral midline incision was made, and the entire colon was removed and placed in a Sylgard-lined dish that was perfused with Krebs-Ringer buffer solution. The region of interest was carefully pinned mucosal side up and a camel-hair paintbrush was used for mucosal stimulation, where each stimulation consisted of a sequence of five brush strokes in approximately 5 seconds. Electronic field stimulation was applied at 20 hertz for 10 seconds; 6 volts .01 millisecond pulse duration. Functional imaging was performed on a Nikon Eclipse FN1 upright fluorescence microscope using Nikon Plan Fluor 20x lens. Image sequences were captured using a Photometrics Prime 95B sCMOS camera and captured on a Windows-based PC using Nikon NIS Elements 4.1. Image sequences were recorded at 25 frames per second.

Completeness: This dataset is complete.

Subjects & Samples: Male (n=11) and female (n=8) adult transgenic mice were used in this study.

Primary vs derivative data: The primary data is organized in folders by subject ID. Each sample subfolder contains .avi microscope movie of calcium transient and movement responses of cholinergic and nitrergic neurons in the middle colon of male and female mice to oral or anal mucosal or electrical field stimulation. There is no derivative data folder.