Figure 1

Enteric neurons produce TNF-α in response to LPS stimulation. (A) rENSpc were treated in a time- and dose-dependent manner with LPS. Quantification of TNF-α secretion was measured by ELISA. Values represent the mean ± SEM of between three and seven independent samples per condition (two-way ANOVA test followed by a Bonferroni post-hoc test; *P <0.05 as compared with the same time point without LPS). (B) Representative images of TNF-α localization in the rENSpc (four independent samples). Immunocytochemical triple labeling of ENS cultures were performed using anti-TNF-α, anti-S100β (glial marker) and anti-Tuj (neuronal marker) antibodies. Examples of neurons expressing TNF-α are depicted with white arrowheads. Scale bar: 50 μm. (C) LPS treatment of enteric glial cell cultures did not induce TNF-α production (between four and ten independent experiments). EGC, enteric glial cells; LMMP, longitudinal muscle/myenteric plexus; LPS, lipopolysaccharides; rENSpc, rat enteric nervous system primary culture; S100β, S100 calcium binding protein beta; SEM, standard error of the mean; TNF-α, tumor necrosis factor alpha; Tuj, βIII-tubulin.