Expression Analyses of Polo-Like Kinase 4, a Gene Product Responsible for Autosomal Recessive Microcephaly and Seckel Syndrome, during Mouse Brain Development
Expression Analyses of Polo-Like Kinase 4, a Gene Product Responsible for Autosomal Recessive Microcephaly and Seckel Syndrome, during Mouse Brain Development
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DOI：
10.1159/000526914
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发表时间：
2022-09
影响因子：
2.9
通讯作者：
Nanako Hamada;I. Iwamoto;M. Noda;M. Nishikawa;K. Nagata
中科院分区：
文献类型：
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作者：
Nanako Hamada;I. Iwamoto;M. Noda;M. Nishikawa;K. Nagata
polo样激酶4 （Plk4）是一种丝氨酸/苏氨酸激酶，在细胞周期的中心粒复制中起核心作用。PLK4基因异常是常染色体隐性绒毛膜视网膜病变-小头畸形综合征和Seckel综合征的原因。在这项研究中，我们通过关注小鼠大脑发育进行了Plk4的表达分析。Western blotting分析显示，分子质量为~ 100 kDa的Plk4在具有特定亚细胞分布的成年小鼠组织中广泛表达。在中枢神经系统中，Plk4在整个发育过程中均有表达，P15后表达量急剧增加，表明Plk4在神经元分化过程中起重要作用。在胚胎14天的小鼠大脑免疫组织化学分析中，Plk4主要在脑室区神经元祖细胞的细胞接触部位检测到。P7时，Plk4在皮质神经元细胞体中弥漫性分布，P30时，Plk4在大脑皮层和海马的兴奋性神经元的神经细胞和胞体以及小脑的浦肯野细胞中富集。值得注意的是，生化分离分析发现Plk4在突触后密度部分富集。免疫荧光分析显示，Plk4与兴奋性突触标志物PSD95和突触素在分化的原代培养海马神经元中部分共定位。这些结果表明Plk4参与了分化神经元突触功能的调控。
Polo-like kinase 4 (Plk4) is a ser/thr kinase, which plays a central role in centriole duplication during the cell cycle. PLK4 gene abnormalities are responsible for autosomal recessive chorioretinopathy-microcephaly syndrome and Seckel syndrome. In this study, we performed expression analyses of Plk4 by focusing on mouse brain development. Western blotting analyses revealed that Plk4 with a molecular mass of ∼100 kDa was broadly expressed in adult mouse tissues with specific subcellular distribution. As to the central nervous system, Plk4 was expressed throughout the developmental process with drastic increase after P15, suggesting an essential role of Plk4 in differentiated neurons. In immunohistochemical analyses with mouse brain at embryonic day 14, Plk4 was detected dominantly at the cell-cell contact sites of neuronal progenitors in the ventricular zone. Plk4 was then diffusely distributed in the cell body of cortical neurons at P7, while it was enriched in the neuropil as well as soma of excitatory neurons in the cerebral cortex and hippocampus and Purkinje cells in the cerebellum at P30. Notably, biochemical fractionation analysis found an enrichment of Plk4 in the postsynaptic density fraction. Then, immunofluorescent analyses showed partial co-localization of Plk4 with excitatory synaptic markers, PSD95 and synaptophysin, in differentiated primary cultured hippocampal neurons. These results suggest that Plk4 takes part in the regulation of synaptic function in differentiated neurons.