Clock genes regulate neurogenic transcription factors, including NeuroD1, and the neuronal differentiation of adult neural stem/progenitor cells
Clock genes regulate neurogenic transcription factors, including NeuroD1, and the neuronal differentiation of adult neural stem/progenitor cells
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DOI：
10.1016/j.neuint.2008.12.005
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发表时间：
2009-05-01
影响因子：
4.2
通讯作者：
Wada, Keiji
中科院分区：
文献类型：
--
作者：
Kimiwada, Tomomi;Sakurai, Mikako;Wada, Keiji
生物钟系统在我们的身体中发挥着多种作用，生物钟基因在不同的大脑区域表达，包括外侧脑室下区 (SVZ)，神经干/祖细胞 (NSPC) 持续存在，出生后神经发生也在此处继续。然而，成年 NSPC 中时钟基因的功能尚不清楚。在这里，我们首先通过免疫组织化学研究了 SVZ 中 Clock 和 Bmal1 的表达模式，然后使用定量 RT-PCR 验证了 17 个时钟和时钟相关基因的表达水平在培养的成体 NSPC 分化过程中如何变化。最后，我们利用RNAi观察了Clock和Bmal1对神经元分化的影响。我们的结果显示，Clock 和 Bmal1 在 SVZ 中表达，并用神经祖细胞标记物 Nestin 和神经干标记物 GFAP 进行双染色。在培养的成年 NSPC 中，时钟基因在分化过程中改变了表达模式，有趣的是，Bmal1 开始内源性振荡。此外，通过RNAi沉默Clock或Bmal1基因会降低神经元标记Map2阳性细胞的百分比和NeuroD1 mRNA的表达水平。这些发现表明，时钟基因参与成人 NSPC 的神经元分化，并可能扩展我们对与成人神经发生和昼夜节律相关的各种神经/心理疾病的理解。 (C) 2008 Elsevier Ltd. 保留所有权利。
The circadian clock system plays multiple roles in our bodies, and clock genes are expressed in various brain regions, including the lateral subventricular zone (SVZ) where neural stem/progenitor cells (NSPCs) persist and postnatal neurogenesis continues. However, the functions of clock genes in adult NSPCs are not well understood. Here, we first investigated the expression patterns of Clock and Bmal1 in the SVZ by immunohistochemistry and then verified how the expression levels of 17 clock and clock-related genes changed during differentiation of cultured adult NSPCs using quantitative RT-PCR. Finally, we used RNAi to observe the effects of Clock and Bmal1 on neuronal differentiation. Our results revealed that Clock and Bmal1 were expressed in the SVZ and double-stained with the neural progenitor marker Nestin and neural stem marker GFAP. In cultured adult NSPCs, the clock genes changed their expression patterns during differentiation, and interestingly, Bmal1 started endogenous oscillation. Moreover, gene silencing of Clock or Bmal1 by RNAi decreased the percentages of neuronal marker Map2-positive cells and expression levels of NeuroD1 mRNA. These findings suggest that clock genes are involved in the neuronal differentiation of adult NSPCs and may extend our understanding of various neurological/psychological disorders linked to adult neurogenesis and circadian rhythm. (C) 2008 Elsevier Ltd. All rights reserved.