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. 2022 Aug 19;164(3):625–637. doi: 10.1097/j.pain.0000000000002750

Figure 2.

Figure 2.

CRD stimulation induces a sharp increase in calcium signals in LSV neurons. (A) Schematic of the recording system for evaluating calcium signals with fiber photometry and visceral pain behavior evoked by CRD stimuli (top) and representative image of GCaMP6f expression in LSV (bottom). Scale bar, 50 μm. (B and C) Heatmap and average Ca2+ transients of LSV neurons in CON and NMD mice receiving CRD stimulation at 20, 40, 60, and 80 mm Hg, respectively. (D) The area under the curve of calcium activity of LSV neurons in CON and NMD mice receiving CRD stimulation at 20, 40, 60, and 80 mm Hg, respectively (F (3, 15) = 82.26, *P < 0.05, ***P < 0.001, two-way repeated-measure ANOVA followed by Bonferroni post hoc test, n = 6 mice per group). € Averaged peak ΔF/F of calcium activity of LSV neurons in CON and NMD mice receiving CRD stimulation at 20, 40, 60, and 80 mm Hg, respectively (F (3, 15) = 113.8, ***P < 0.001, two-way repeated-measure ANOVA followed by Bonferroni post hoc test, n = 6 mice per group). (F) Schematic of the recording system for evaluating calcium signals with fiber photometry and somatic stimulation evoked by von Frey fiber, tail pinch, and tail brush. (G and H) Heatmap and average Ca2+ transients of LSV neurons in CON and NMD mice receiving tail brush, tail pinch, and von Frey filament (Crl and Ipl) stimulation, respectively. (I) The area under the curve of calcium activity of LSV neurons in CON and NMD mice receiving tail brush, tail pinch, and von Frey filament (Crl and Ipl) stimulation, respectively (F (3, 20) = 0.5363, P > 0.05, two-way ANOVA followed by Bonferroni post hoc test, n = 6 mice per group). (J) Averaged peak ΔF/F of calcium activity of LSV neurons in CON and NMD mice receiving tail brush, tail pinch, and von Frey filament (Crl and Ipl) stimulation, respectively (F (3, 20) = 0.7553, P > 0.05, two-way ANOVA followed by Bonferroni post hoc test, n = 6 mice per group). Crl, contralateral; Ipl, ipsilateral. n.s. indicates nonsignificant differences, P > 0.05. See also Figure S1, available at http://links.lww.com/PAIN/B695. ANOVA, analysis of variance; CRD, colorectal distention; LSV, lateral septal ventral; NMD, neonatal maternal deprivation.