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首页> 外文会议>Meeting on Cell Signaling World: Signal Transduction Pathways as Therapeutic Targets >Intensive Remodeling of Purkinje Cell Spines after Climbing Fibers Deafferentation Does Not Involve MAPK and Akt Activation
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Intensive Remodeling of Purkinje Cell Spines after Climbing Fibers Deafferentation Does Not Involve MAPK and Akt Activation

机译：攀登纤维脱除咖啡因后的浦肯野细胞棘的密集重塑不涉及MAPK和Akt激活

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Subtotal lesion of the inferior olive (IO) achieved by treat ing experimental animals with 3-acetylpyridine (3AP) induces partial Purkinje cells (PCs) deafferentation that leads to PC hyperactivity and new spine formation. Coincidentally, the olivary terminals belonging to the few survived olivary neurons undergo an extensive collateral sprout ing resulting in reinnervation of the neighboring denervated PCs. We obtained chemical deafferentation of PCs in adult rats (body weight, 120-170 g; age, 35-40 days) by a single intraperitoneal injection of 3AP (65 mg/kg body weight), and as early as 3 days after 3AP treatment, important morphological changes could be observed on PCs. Mitogen activated protein kinase (MAPK) cascades and more specifically extra cellular signal-regulated kinases 1/2 (ERK1/2) play a critical role in the signaling events underlying synaptic plasticity. For instance, long-term depression (LTD) in the adult hippocampus and long-term potentiation (LTP) in cerebellum both involve ERK activation. Since PCs deprived of their climbing fibers (CFs) afferents initiate an intensive remodel ing of the spines and rapid recall of the remaining CFs, it prompted us to see whether the observed phenomena correlated with MAPK and Akt activation. Immunohistochemistry and Western blotting were done at various time points after 3AP application (from 24 h to 6 days), as the exact dynamics of CF loss is not precisely known. As judged by Western blotting, there was no increase of activated ERK in the cere bellum. However, immunohistochemistry revealed increased ERK phos phorylation in the "pinceaux" of basket cells in 3AP animals. Similarly, stress-activated protein kinase (SAPK)/c-Jun N-terminal kinase (JNK), p38 MAPK, and Akt activation were also studied by means of Western blotting and immunohistochemistry. Upon 3AP treatment no changes in phosphorylation status could be seen in the different kinases subjected to analysis. Our results suggest that activation of MAPK and Akt cascades is not essential in this model of neuronal plasticity.
机译：通过用3-乙酰基吡啶（3AP）处理实验动物而获得的下橄榄（IO）的小计病变可引起部分Purkinje细胞（PCs）脱除咖啡因，从而导致PC机能亢进和新的脊柱形成。巧合的是，属于少数存活的橄榄神经元的橄榄末端经历了广泛的附带发芽，从而导致邻近的失神经的PC的神经支配。我们通过单次腹膜内注射3AP（65 mg / kg体重）以及早在3AP治疗后3天，获得了成年大鼠（体重120-170 g;年龄，35-40天）中PC的化学除草作用，可以在PC上观察到重要的形态变化。丝裂原活化蛋白激酶（MAPK）级联，更具体地讲，细胞外信号调节激酶1/2（ERK1 / 2）在突触可塑性潜在的信号传导事件中起关键作用。例如，成年海马的长期抑郁（LTD）和小脑的长期增强（LTP）均涉及ERK激活。由于PC失去了攀爬纤维（CF）的传入，因此开始了棘突的重塑并迅速召回了剩余的CF，这促使我们看到观察到的现象是否与MAPK和Akt激活相关。免疫组化和蛋白质印迹在3AP施用后的不同时间点（从24小时到6天）进行，因为CF丢失的确切动态尚不清楚。通过蛋白质印迹判断，在小脑中活化的ERK没有增加。但是，免疫组织化学显示3AP动物篮子细胞的“ pinceaux”中ERK磷酸化增加。同样，应力激活蛋白激酶（SAPK）/ c-Jun N端激酶（JNK） 还通过蛋白质印迹和免疫组织化学研究了p38 MAPK和Akt激活。经3AP处理后，在进行分析的不同激酶中未观察到磷酸化状态的变化。我们的结果表明，在这种神经元可塑性模型中，MAPK和Akt级联反应的激活不是必需的。

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