{"id": "package:b98dfc6a-1104-4c27-bbbb-bc02d5724bc6", "name": "ISP_20190913_G1.xlsx", "self_uri": "https://services.scicrunch.io/sparc/drs/v1/objects/b98dfc6a-1104-4c27-bbbb-bc02d5724bc6", "size": 527351, "created_time": "2020-10-16T13:34:34,135904Z", "updated_time": "2022-12-13T15:43:03,032359Z", "version": "2", "mime_type": "application/vnd.openxmlformats-officedocument.spreadsheetml.sheet", "checksums": [{"checksum": "9f55335260d4beb8fa10fee17af3ae7c2922ce18d90f527f80e2d37752557e33", "type": "sha256"}], "access_methods": [{"type": "s3", "access_url": {"url": "s3://prd-sparc-discover50-use1/124/files/primary/stretch/sub-20190913_G1/sam-20190913_G1/ISP_20190913_G1.xlsx"}, "region": "us-east-1"}], "dataset": {"id": "124", "doi": "DOI:10.26275/0khe-2os4", "title": "In vitro imaging of mechanosensitive submucous neurons in the porcine colon", "description": "This dataset is about electrophysiological experiment on porcine colonic submucosal enteric neurons. The neurons were challenged with 2 different mechanical stimuli (compression and stretch). Neuronal responses to the stimuli were recorded and analyzed.", "abstract": "In this study we investigated the sensitivity of enteric neurons to mechanical stimuli comparing tissue samples from porcine proximal and distal colon. Tissue samples were taken from apparently healthy pigs (Leine-Fleisch GmbH, Laatzen, Germany), placed in ice-cold oxygenated Krebs solution for preparation and immediately transferred to our laboratory. Tissues were then dissected in ice-cold oxygenated Krebs solution for preparation to obtain whole-mount inner submucosal plexus preparations. Samples were used for ultrafast neuroimaging technique combined with a voltage sensitive dye (VSD) as well as for immunohistochemistry.\nIn order to stain selected submucosal ganglia, VSD Di-8-ANEPPS (20 μM) was applied directly into the ganglion at moderate pressure (≤ 0.5 bar). The viability of ganglia was proven by applying 100 μM nicotine on the surface of a ganglion. Mechanical stimulation included compression and tension of submusosal neurons. Compression was induced by intraganglionic volume injection of experimental Krebs solution at 0.5 bar for 500 ms. Tension of entire ganglia was performed bidirectionally by moving a self-constructed stretching tool in opposite directions perpendicular to the ganglion axis. The preparations were examined with an inverted microscope equipped with an appropriate filter set. Pictures were acquired with a CMOS camera connected to a computer and controlled by Turbo SM 64 Software.\nWe used primary antibodies against Hu, Chat, SP and NOS. Specimens were first incubated in Triton X-100 (0.5%)/PBS/NaN3 (0.1%)/horse serum (4%) for 1 h at room temperature followed by 12h incubation with the primary antibodies, respectively. After incubation for 2 h at room temperature with the respective secondary antibodies specimens were washed in PBS and cover slipped with a solution of PBS/NaN3 containing 65% glycerol. The preparations were examined with an epifluorescence microscope equipped with appropriate filter blocks. Pictures were acquired with a camera connected to a computer and controlled by Olympus cellSens Standard Software.\nAnalysis was performed on Excel tables. The approximate size of the whole data set is around 1 GB."}}