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Data to support: Functional characterization of ovine dorsal root ganglion neurons reveals peripheral sensitization after osteochondral defect


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Authors

Chakrabarti, Sampurna 
Wong, Katherine 
Newell, Karin 

Description

All the raw data from electrophysiology and Ca2+ imaging present in Figure 1, Figure 2 and Supplementary Figure 1 of the paper.

Electrophysiology data were collected from sheep dorsal root ganglion neurones, as described in detail in the paper, on a HEKA EPC 10 amplifier, action potential threshold, half peak duration, amplitude, after-hyperpolarization duration and amplitude were measured using FitMaster (HEKA) software or IgorPro software (Wavemetrics).

For Ca2+-imaging, neurons were loaded with Fluo-4 AM (as described in the paper) and dishes placed on microscope (Nikon Eclipse Tie–S, Nikon) for imaging. Fluo-4 fluorescence was excited by a 470 nm LED (Cairn Research) and images were captured by a digital camera(Zyla cSMOS, Andor) at 1 Hz with 50-ms exposure time using Micro-Manager software (v1.4; NIH). As detailed in the paper: KCl positive cells and one black background were drawn manually as a region of interest (ROI) using ImageJ software and the mean gray value of selected ROIs in sequence was extracted. Extracted data were then analyzed by lab-developed R toolbox (https://github.com/amapruns/Calcium-Imaging- Analysis-with-R) to calculate the change in Ca21 influx (normalized to peak KCl response (DF/Fmax) with back- ground subtraction) and the percentage of agonist re- spondent cells (cells with DF/Fmax value ,0.001 and peak after 30 s were deleted manually).

Version

Software / Usage instructions

Files can be opened with GraphPad Prism and Igor is required to open files with voltage-gated ion channel electrophysiology data.

Keywords

Electrophysiology, Imaging

Publisher

Sponsorship
S.C. was supported by the Gates Cambridge Trust scholarship. This work was supported by funding from Versus Arthritis (RG21973) and BBSRC (BB/R006210/1) to E.St.J.S and Horizon 2020 (RG90905) and Innovate UK (RG87266) to F.M.D.H.
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