PDK1 Regulates the Maintenance of Cell Body and the Development of Dendrites of Purkinje Cells by pS6 and PKCγ
PDK1 Regulates the Maintenance of Cell Body and the Development of Dendrites of Purkinje Cells by pS6 and PKCγ
复制标题
PDK1通过pS6和PKCγ调节浦肯野细胞胞体的维持和树突的发育
DOI：
10.1523/jneurosci.2496-19.2020
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发表时间：
2020-07-15
影响因子：
5.3
通讯作者：
Gao, Jun
中科院分区：
文献类型：
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作者：
Liu, Rui;Xu, Min;Gao, Jun
3-磷脂酰肌醇依赖的蛋白激酶-1(PDK1)在哺乳动物脑发育过程中发挥着重要作用。在这里，我们通过将pv-cre或pcp2-cre小鼠与pDK1(f1/f1)小鼠杂交，产生pDK1条件性基因敲除小鼠(CKO)，研究了pDK1在浦肯野细胞(PC)中的作用。雄性小鼠用于行为学测试，其他实验在不同性别的小鼠上进行。这些PDK1-CKO小鼠的小脑体积变小，运动平衡和协调性受损。通过电生理记录，我们观察到PDK1-CKO小鼠小脑脑片PC自发放电的减少。此外，与对照组相比，PDK1-CKO组小鼠的PC胞体大小呈时间依赖性地减小。PDK1缺失后，PC的形态复杂性也降低。MTOR1激动剂3-苄基-5-((2-硝基苯氧基)甲基)-二氢呋喃-2(~H)-1上调pS6后，pDK1-CKO小鼠rpS6(还原核糖体蛋白S6)磷酸化水平降低和PKC-γ表达降低，部分挽救了PC胞体的缩小，小脑注射重组腺相关病毒-PKC-γ挽救了PDK1-CKO小鼠树突状突起的复杂性。综上所述，我们的数据表明，PDK1通过调节rpS6的磷酸化和PKCγ的表达，控制PC的细胞体维持和树突的发育，对小脑运动协调至关重要。
3-Phosphoinositide-dependent protein kinase-1 (PDK1) plays a critical role in the development of mammalian brain. Here, we investigated the role of PDK1 in Purkinje cells (PCs) by generating the PDK1-conditional knock-out mice (cKO) through crossing PV-cre or Pcp2-cre mice with Pdk1(fl/fl) mice. The male mice were used in the behavioral testing, and the other experiments were performed on mice of both sexes. These PDK1-cKO mice displayed decreased cerebellar size and impaired motor balance and coordination. By the electrophysiological recording, we observed the reduced spontaneous firing of PCs from the cerebellar slices of the PDK1-cKO mice. Moreover, the cell body size of PCs in the PDK1-cKO mice was time dependently reduced compared with that in the control mice. And the morphologic complexity of PCs was also decreased after PDK1 deletion. These effects may have contributed to the reduction of the rpS6 (reduced ribosomal protein S6) phosphorylation and the PKC gamma expression in PDK1-cKO mice since the upregulation of pS6 by treatment of 3-benzyl-5-((2-nitrophenoxy) methyl)-dihydrofuran-2(3H)-1, the agonist of mTOR1, partly rescued the reduction in the cell body size of the PCs, and the delivery of recombinant adeno-associated virus-PKC gamma through cerebellar injection rescued the reduced complexity of the dendritic arbor in PDK1-cKO mice. Together, our data suggest that PDK1, by regulating rpS6 phosphorylation and PKC gamma expression, controls the cell body maintenance and the dendritic development in PCs and is critical for cerebellar motor coordination.