Activation of the CREB/c-Fos Pathway during Long-Term Synaptic Plasticity in the Cerebellum Granular Layer.
Activation of the CREB/c-Fos Pathway during Long-Term Synaptic Plasticity in the Cerebellum Granular Layer.
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DOI：
10.3389/fncel.2017.00184
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发表时间：
2017
影响因子：
5.3
通讯作者：
D'Angelo E
中科院分区：
文献类型：
--
作者：
Gandolfi D;Cerri S;Mapelli J;Polimeni M;Tritto S;Fuzzati-Armentero MT;Bigiani A;Blandini F;Mapelli L;D'Angelo E
长期增强和抑制（LTP和LTD）的诱导被认为触发基因表达和蛋白质合成，导致突触和神经元变化的巩固。然而，尽管LTP和LTD被认为在小脑颗粒层的感觉-运动学习中发挥重要作用，但它们与这些机制的关联尚不清楚。在这里，我们研究了在急性小脑切片中，脑电波爆发刺激（TBS）诱导突触可塑性后，camp反应元件结合蛋白（CREB）的磷酸化和即时早期基因c-Fos通路的激活。用电压敏感染料成像（VSDi）定位LTP和LTD。在TBS后的两个时间点（15 min和120 min），对应于可塑性的早期和晚期，将切片固定并处理，以评估CREB磷酸化（P-CREB）和c-FOS蛋白水平，以及CREB和c-FOS mRNA表达。高水平的P-CREB和Creb/c-Fos在c-Fos之前被检测到，如果Creb磷酸化触发基因表达，然后是蛋白质合成，这是预期的。在n -甲基- d -天冬氨酸受体（NMDAR）拮抗剂存在的情况下，对照切片和经TBS刺激的切片之间没有差异。有趣的是，CREB/c-Fos系统的激活显示出与长期突触可塑性相关的共定位程度。这些结果表明，在小脑输入阶段，nmdar依赖的可塑性与转录和转录后过程有关，可能有助于小脑的学习和记忆巩固。
The induction of long-term potentiation and depression (LTP and LTD) is thought to trigger gene expression and protein synthesis, leading to consolidation of synaptic and neuronal changes. However, while LTP and LTD have been proposed to play important roles for sensori-motor learning in the cerebellum granular layer, their association with these mechanisms remained unclear. Here, we have investigated phosphorylation of the cAMP-responsive element binding protein (CREB) and activation of the immediate early gene c-Fos pathway following the induction of synaptic plasticity by theta-burst stimulation (TBS) in acute cerebellar slices. LTP and LTD were localized using voltage-sensitive dye imaging (VSDi). At two time points following TBS (15 min and 120 min), corresponding to the early and late phases of plasticity, slices were fixed and processed to evaluate CREB phosphorylation (P-CREB) and c-FOS protein levels, as well as Creb and c-Fos mRNA expression. High levels of P-CREB and Creb/c-Fos were detected before those of c-FOS, as expected if CREB phosphorylation triggered gene expression followed by protein synthesis. No differences between control slices and slices stimulated with TBS were observed in the presence of an N-methyl-D-aspartate receptor (NMDAR) antagonist. Interestingly, activation of the CREB/c-Fos system showed a relevant degree of colocalization with long-term synaptic plasticity. These results show that NMDAR-dependent plasticity at the cerebellum input stage bears about transcriptional and post-transcriptional processes potentially contributing to cerebellar learning and memory consolidation.