Fig. 3: Lung-innervating sensory neurons mediate the activation of 5-HTDRN neurons by peripheral MSCs. | Nature Communications

Fig. 3: Lung-innervating sensory neurons mediate the activation of 5-HTDRN neurons by peripheral MSCs.

From: Mesenchymal stromal cells alleviate depressive and anxiety-like behaviors via a lung vagal-to-brain axis in male mice

Fig. 3

a Representative images from three independent experiments showing GFP-MSC distribution. Scale bar, 200 µm. b Quantification of GFP-MSCs. One-way ANOVA: F(4, 10) = 303.6 **** P = 0.00000000022. n = 3 mice. c Label design for mCherry-GFP-MSCs. d Representative image from three independent experiments showing that GFP-MSCs co-expressed mCherry. Scale bar, 10 µm. e Representative image from three independent experiments showing the colocalization between mCherry secreted from MSCs and pulmonary nerves (TUBB3). Scale bar, 50 µm. f Representative image showing from three independent experiments the colocalization between mCherry secreted from MSCs and pulmonary vagal nerves (VGLUT2). Scale bar, 50 µm. g Representative sequential images from three independent experiments displaying calcium signal response to MSC suspensions inside the ex-vivo lungs of VGLUT2-GCaMP6 mice. Arrow points to a region showing an increase in GCaMP6 fluorescence. Scale bar, 10 µm. n = 3 mice. Right, real-time changes in fluorescence intensity were expressed as percentage changes over baseline ((F-F0)/F0). h Representative images and statistical analysis showing the expression of c-Fos in nodose ganglia. Scale bar, 200 µm. Two-tailed t test: t = 4.685, df = 8 ** P = 0.0016. n = 5 mice. i Representative images from three independent experiments showing the NTS c-Fos expression. Scale bar, 200 µm. AP, area postrema. j The experimental procedures. k Representative images from three independent experiments showing the NTS c-Fos expression. Scale bar, 200 µm. l Statistical analysis of the NTS c-Fos expression. Brown-Forsythe ANOVA test: ** P = 0.0046. n = 5 mice from three independent experiments. m Representative images showing the colocalization of 5-HT and c-Fos in the DRN. Scale bar, 100 µm. aq, mesencephalic aqueduct. n Statistical analysis of the DRN c-Fos expression. One-way ANOVA: F(3, 16) = 12.32 *** P = 0.00020. n = 5 mice. o Statistical analysis of the colocalization of 5-HT and c-Fos. One-way ANOVA: F(3, 33) = 8.545 *** P = 0.00024. n = 5, 7, 12, 13 mice separately. Illustrations created with BioRender.com. Source data are provided as a Source Data file.

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