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Fig. 4 | Acta Neuropathologica Communications

Fig. 4

From: Sphingosine-1-phosphate receptor inhibition prevents denervation-induced dendritic atrophy

Fig. 4

Change in Sphingosine-1-phosphate (S1P) and S1P receptor (S1PR) 1 and 3 mRNA levels following entorhinal denervation in vitro. a Laser microdissection (LMD) was employed to collect tissue from the granule cell layer (GCL) the inner molecular layer (IML) and the outer molecular layer (OML) of denervated cultures (at 2, 7 and 14 days post lesion; dpl) and non-denervated cultures (control; age- and time-matched to denervated cultures). Scale bar: 50 μm. b, c LMD/qPCR results of non-denervated and denervated cultures. b RNA integrity numbers (RIN) for the acquired samples. S1PR1- and S1P3-mRNA levels were assessed in the isolated tissue using qPCR. c An increase in S1PR1-mRNA was observed in the OML at 7 dpl. Similarly, increased S1PR3-mRNA levels were observed in the OML (at 2 dpl). S1PR1- and S1PR3-mRNA levels in the IML and GCL were not significantly changed (data not shown; n = 4–9 cultures per group; Kruskal-Wallis test followed by Dunn’s multiple comparisons test: *, p <0.01; ns, not significant; age- and time-matched non-denervated controls were not significantly different and therefore pooled). d Mass spectrometry disclosed a gradual increase in S1P levels in denervated hippocampal tissue as compared to samples taken from age- and time-matched non-denervated cultures (control; n = 9–11 independent experiments per group; Kruskal-Wallis test followed by Dunn’s multiple comparisons test: **, p <0.01; ns not significant; age- and time-matched non-denervated controls were not significantly different and therefore pooled)

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