Fig. 4: BDNF from astrocytes regulates neural network levels of activity in an in vitro model of temporal lobe epilepsy. | Cell Death & Disease

Fig. 4: BDNF from astrocytes regulates neural network levels of activity in an in vitro model of temporal lobe epilepsy.

From: Astrocytic BDNF and TrkB regulate severity and neuronal activity in mouse models of temporal lobe epilepsy

Fig. 4

a Representative average image of a hippocampal neurons-astrocytes co-culture at 15 DIV loaded with Fluo4-AM. b Calcium fluorescence traces from individual neurons highlighted in a. Black vertical lines indicate inferred spikes from the three neurons. c Experimental design: after 10 min recording in basal conditions, 4-AP is added to the culture and after 5 min of drug diffusion and stabilization, and neuronal activity is recorded for additional 10 min. d 4-AP treatment increased the firing rate of individual neurons in both conditions; cultured with BDNF+/+ or BDNF−/− astrocytes (two-way ANOVA, 4-AP effect: F(1,22) = 17.23, p < 0.001), indicating the effect of the treatment increasing neuronal activity. e 4-AP treatment has differential effect on the number of active neurons depending on if they were co-cultured with BDNF+/+ or BDNF−/− astrocytes (two-way ANOVA, interaction effect: F(1,26) = 5.204, p < 0.05; 4-AP effect: F(1,26) = 6.419, p < 0.05); genotype effect: F(1,26) = 8.881, p < 0.01). Post hoc analysis revealed a significant increase in the number of active neurons co-cultured with BDNF+/+ astrocytes after addition of 4-AP, but not in the ones co-cultured with BDNF−/− astrocytes; after 4-AP treatment, the number of active neurons was different between BDNF+/+ and BDNF−/− co-cultures. f 4-AP treatment increases the global firing rate in neurons in both conditions (two-way ANOVA, 4-AP effect: F(1,26) = 7.266, p < 0.05); genotype effect: F(1,26) = 5.654, p < 0.05). Post hoc analysis showed significance only in neurons co-cultured with BDNF+/+ astrocytes, but not in neurons co-cultured with BDNF−/− astrocytes. Each point represents the average value of an individual experiment. Black lines show the mean of the group (n = 10 BDNF+/+ basal; 10 BDNF−/− basal; 5 BDNF+/+ 4-AP; 5 BDNF−/− 4-AP). g 4-AP treatment increased the firing rate in both conditions; individual neurons co-cultured with WT or pGFAP-BDNF astrocytes (two-way ANOVA, 4-AP effect: F(1,40) = 22.17, p < 0.0001; genotype effect: F(1,40) = 4.77, p < 0.05). h 4-AP treatment increased the number of active neurons in both conditions; in WT neurons co-cultured with WT or pGFAP-BDNF astrocytes (two-way ANOVA, 4-AP effect: F(1,40) = 13.57, p < 0.001). i 4-AP treatment increased the global firing rate in both conditions; neurons co-cultured with WT or pGFAP-BDNF astrocytes (two-way ANOVA, 4-AP effect: F(1,40) = 31.75, p < 0.0001). Each point represents the average value of an individual experiment. Black lines show the mean of the group (n = 12WT basal; 11pGFAP-BDNF basal; 11WT 4-AP; 11 BDNF−/− 4-AP). Data were analyzed by two-way ANOVA followed by Bonferroni’s post hoc test. *p < 0.05, **p < 0.01, ***p < 0.001 compared to WT basal; $$p < 0.001 compared to BDNF−/− basal; #p < 0.05, ###p < 0.001 compared to pGFAP-BDNF. Scale bar = 200 μm.

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