Study purpose: The main purpose of the study is to examine the distribution of nitergic and cholinergic neurons in the colon.
Data collection: Tissue samples from the colon were collected in transgenic mice to monitor and drive the activity of cholinergic and nitergic neurons. GCaMP3-expressing myenteric neurons were identified by fluorescence on an inverted microscope (Keyence, Itaska, IL) and counted in 500-μm2 areas across the entire colonic circumference of each colonic segment. Data consist of multiple stacked and reconstructed images (bcf and tif format) of cholinergic, nitergic, and glial distribution.
Primary conclusion: None stated
Experimental Design: Mice were into allocated into 3 experimental groups based on targeted neurons: ChAT (3 female and 4 male mice; cholinergic neurons), (2) nNOS (3 female, 4 male mice; nitergic neurons), and (3) Wnt-1 (4 female and 4 male mice; enteric neurons and glia cells). The researchers choose to label cells of interest with the following transgenic approach: GCaMP3 mice (RRID:IMSR_JAX:029043) were crossed with these Cre specific drivers: ChAT-Cre (RRID:MMRRC_037336), nNOS-CreERT2 (RRID:IMSR_JAX:014541), and Wnt-1Cre2 (RRID:IMSR_JAX:022501) which drive expression, upon tamoxifen injection, in nitrergic enteric neurons. Imaging was automated using Keyence BZ-X automated microscope at 20X magnification.
Subjects & Samples: Adult male and female C57BL/6 transgenic mice between 47 and 116 days old were used. Samples were taken from the distal, middle, and proximal regions of the colon for each subject.
Primary vs derivative data: The primary folder contains folders with raw data for each subject, divided by region of colon as a zip file. The zip file consists of a bcf file and tif stack files. The derived folder contains reconstructed images for each subject, by colon region as tif files. Image data (.jp2) was derived from derivative distribution map images (.tif). Images were converted with 40:1 compression to .jp2 by MBF Bioscience for web streaming and visualization on the SPARC Data Portal.
Important notes: (1) The docs folder contains a text document with the image scale measurements, which is 1 pixel = 0.751 um. (2) Some of the zip files in the primary folder are extremely large and may not open on a local drive. The average file size: 14.2 GB (3) Mice. Conditional GCaMP3 (RRID:IMSR_JAX:029043) or channelrhodopsin-2 (RRID:IMSR_JAX:024109) mice, which allow for cell-specific expression of the optical sensor GCaMP3 or the optical actuator ChR2, were crossed to ChAT-Cre mice (RRID:MMRRC_037336), which drive expression in cholinergic enteric neurons; and nNOS-CreERT2 mice (RRID:IMSR_JAX:014541), which drive expression, upon tamoxifen injection, in nitrergic enteric neurons. For convenience, these are referred to as ChAT-GCaMP3 or nNOS-GCaMP3/ChR2 mice.
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