Promotion of dendritic differentiation of cerebellar Purkinje cells by Ca2+/calmodulin-dependent protein kinase IIα, IIβ and IV and possible involvement of CREB phosphorylation
Promotion of dendritic differentiation of cerebellar Purkinje cells by Ca2+/calmodulin-dependent protein kinase IIα, IIβ and IV and possible involvement of CREB phosphorylation
复制标题
Ca2+/钙调蛋白依赖性蛋白激酶 IIα、IIβ 和 IV 促进小脑浦肯野细胞的树突分化，并可能涉及 CREB 磷酸化
DOI：
10.1016/j.neuroscience.2021.01.024
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发表时间：
2021
期刊：
影响因子：
3.3
通讯作者：
Masahiko Tanaka
中科院分区：
文献类型：
--
作者：
Yuki Horie;Toshiaki Arame;Naohide Hirashima;Masahiko Tanaka
小脑浦肯野细胞在大脑神经元中发育出最精致的树突树。为了研究Ca2+/钙调素依赖性蛋白激酶（CaMK） IIα， IIβ和IV在浦肯野细胞树突状分化中的作用，我们使用单细胞电穿孔技术将针对这些CaMK的siRNA引入小脑细胞培养的浦肯野细胞。单细胞电穿孔使我们能够将siRNA转移到异质细胞群中的特定细胞中。此外，我们可以简单而可靠地将多种siRNA转移到细胞中，只需将它们装载在一个微移液管中。任何一种抗camkii α， ii β和div的siRNA（单敲除）或任何两种抗这些ecamks的siRNA的组合（双敲除）对浦肯野细胞的树突状分化没有显著影响。然而，这三种siRNA对这些ecamks的联合抑制了浦肯野细胞树突的分支。此外，三重敲除降低了浦肯野细胞中CREB的磷酸化。这些发现提示CaMKIIα， i β和IV促进浦肯野细胞的树突分化，并可能参与CREB磷酸化作为这些camk的共同底物。
Cerebellar Purkinje cells develop the most elaborate dendritic trees among neurons in the brain. To examine the role of Ca2+/calmodulin-dependent protein kinase (CaMK) IIα, IIβ and IV in the dendritic differentiation of Purkinje cells, we introduced siRNA against theseCaMKsinto Purkinje cells in cerebellar cell cultures using a single-cell electroporation technique. Single-cell electroporation enables us to transfer siRNA into specific cells within a heterogeneous cell population. In addition, we can easily and reliably transfer multiple types of siRNA into a cell simply by loading them together in one micropipette. Any one of the siRNA againstCaMKIIα, IIβandIV(single knockdown) or any combinations of two of the siRNA against theseCaMKs(double knockdown) had no significant effects on the dendritic differentiation of Purkinje cells. However, the combination of all three siRNA against theseCaMKs(triple knockdown) inhibited the branching of Purkinje cell dendrites. Furthermore, the triple knockdown reduced the phosphorylation of CREB in Purkinje cells. These findings suggest the promotion of dendritic differentiation of Purkinje cells by CaMKIIα, IIβ and IV and the possible involvement of phosphorylation of CREB as a common substrate of these CaMKs.