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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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Chakrabarti, Sampurna 
Wong, Katherine 
Newell, Karin 


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 ( 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).


Software / Usage instructions

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


Electrophysiology, Imaging


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.