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. 2018 Apr 9;12:224. doi: 10.3389/fnins.2018.00224

Figure 5.

Figure 5

Bicuculline-evoked neuronal activation triggers upregulation and stabilization of Mmp-9 mRNA in hippocampal neurons. (A) Mmp-9 mRNA accumulates in primary rat hippocampal neuronal cultures after neuronal activation. On Day 7 in vitro, neuronal cultures were treated with bicuculline (50 μM). For each analysis, equal amounts of RNA samples isolated from the unstimulated (control) and activated (2 and 4 h after bicuculline) cultures were used. Mmp-9 expression was normalized against 18S rRNA expression. One-way ANOVA was followed by Tukey's multiple comparisons test. Values are mean ± SEM (*p < 0.05; **p < 0.01; ***p < 0.001; n = 6). (B) Bicuculline treatment strongly extends the Mmp-9 mRNA half-life in primary hippocampal neurons. On Day 7 in vitro, actinomycin D (5 μg/mL) was added to the medium of the unstimulated (control) and activated (4 h after bicuculline) hippocampal neuronal cultures for 30 min. Then, RNA was isolated from the cultures at 0, 1, 2, 4, 6, 8, and 12 h after administration of actinomycin D, and Mmp-9 mRNA expression was evaluated by qRT-PCR. Mmp-9 mRNA expression was normalized against 18S rRNA expression. Results are presented as a percentage of the remaining Mmp-9 mRNA transcripts (the amount of Mmp-9 mRNA at Time 0 is assumed to be 100%). Two-way ANOVA followed by Bonferroni's multiple comparisons test. Values are mean ± SEM (**p < 0.01; ***p < 0.001; n = 4).