Extended Data Fig. 2: Specificity of 5-HT sensors.
From: Improved green and red GRAB sensors for monitoring spatiotemporal serotonin release in vivo
Specificity test of indicated sensors in HEK293T cells (a, b) or cultured rat cortical neurons (c–f) to 5-HT alone, 5-HT together with SB, 5-HT together with RS, and 5-HT precursor, 5-HT metabolites, as well as other neurotransmitters and neuromodulators (all compounds at 10 μM except RS at 100 μM). 5-HTP, 5-hydroxytryptophan; 5-HIAA, 5-hydroxyindole acetic acid; DA, dopamine; NE, norepinephrine; HA, histamine; MT, melatonin; OA, octopamine; Glu, glutamate; GABA, gamma-aminobutyric acid; ACh, acetylcholine; Gly, glycine. Norm., normalized. n = 3 wells for each group with 200–500 cells per well. One-way ANOVA followed by Tukey’s multiple-comparison tests, in panel a, F13,28 = 180.2, P = 2.08 × 10−23, post hoc test: P = 0 for 5-HT versus 5-HT and RS, and other compounds; in panel b, F13,28 = 120, P = 5.52 × 10−21, post hoc test: P = 0 for 5-HT versus 5-HT and RS, and other compounds; in panel c, F13,28 = 148.9, P = 2.86 × 10−22, post hoc test: P = 0 for 5-HT versus 5-HT and RS, and other compounds; in panel d, F13,28 = 918, P = 3.16 × 10−33, post hoc test: P = 0 for 5-HT versus 5-HT and RS, and other compounds; in panel e, F13,28 = 44.2, P = 3.65 × 10−15, post hoc test: P = 4.39 × 10−7, 2.06 × 10−7, 9.18 × 10−8, 1.26 × 10−7, 1.26 × 10−7, 1.26 × 10−7, 2.08 × 10−7, 2.08 × 10−7, 1.26 × 10−7, 1.26 × 10−7, 2.09 × 10−7 and 9.18 × 10−8 for 5-HT versus 5-HT and RS, and 5-HTP, 5-HIAA, DA, NE, HA, MT, OA, Glu, GABA, ACh and Gly; in panel f, F13,28 = 87.9, P = 3.83 × 10−19, post hoc test: P = 1.75 × 10−9, 0, 2.33 × 10−11, 0, 0, 0, 0, 0, 0, 0, 0 and 0 for 5-HT versus 5-HT and RS, and 5-HTP, 5-HIAA, DA, NE, HA, MT, OA, Glu, GABA, ACh and Gly. Data are shown as mean ± s.e.m., with the error bars indicating the s.e.m. ***P < 0.001.
