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. 2021 Jan 26;11:2249. doi: 10.1038/s41598-021-81269-6

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Cluster analysis of spontaneous EPSCs in delay and tonic-firing superficial dorsal horn neurons. Three dimensional representations of the sEPSCs in (A1,A2) delay neurons and (B1,B2) tonic neurons plotted based on interevent interval (x-axis), amplitude (y-axis), and charge transfer (z-axis). Mapping of the clustering results and treatment shown for the total sample of sEPSCs. The sEPSCs in each neuron were clustered independently from the other neurons. Optimal partitioning around medoids identified 3 primary clusters present in all neurons in both treatment conditions (naming based on amplitude; interevent interval): small; short, small; long, and large. Two additional clusters were identified in some neurons, but were not present in all neurons in both conditions: small; mid and medium. Neurons sampled: 19 delay (5 control, 14 BDNF); 9 tonic (5 control, 4 BDNF). Total number of sEPSCs: 33,206 in delay, control; 108,261 in delay, BDNF; 40,658 in tonic, control; 36,521 in tonic, BDNF. The effects of BDNF on sEPSC amplitude, charge transfer (i.e., area under the curve), and peak time were assessed for the 3 primary clusters (large cluster shown, refer to Supplemental Fig. 3 for the two small-amplitude clusters). (C1,C2) Treatment with BDNF significantly decreased sEPSC amplitudes in tonic neurons, but a significant effect was not observed in the delay neurons. Cumulative distributions of the measured amplitudes shown (left) alongside the means ± standard error of the means (right). (D1,D2) The BDNF treatment significantly reduced sEPSC charge transfer measures in both delay and tonic neurons. (E1,E2) Interevent intervals for the sEPSCs within the large cluster were significantly decreased in the BDNF-treated neurons. (F1, F2) The sEPSC activation kinetics were significantly faster in the BDNF-treated neurons, but no significant effect was observed for the sEPSCs in the delay neurons. Large sEPSC cluster sample sizes: delay neurons control n = 753, BDNF n = 4539; tonic neurons control n = 2348, BDNF n = 2367. Comparisons between means were made using Brown-Forsythe and Welch’s ANOVA tests and Games-Howell post-hoc test. **P < 0.01, ****P < 0.0001.