A pivotal role of calcineurin signaling in development and maturation of postnatal cerebellar granule cells.
A pivotal role of calcineurin signaling in development and maturation of postnatal cerebellar granule cells.
复制标题
DOI：
10.1073/pnas.0501972102
复制
发表时间：
2005-04
影响因子：
11.1
通讯作者：
Masaaki Sato;Kazunori Suzuki;Hiroshi Yamazaki;S. Nakanishi
中科院分区：
文献类型：
--
作者：
Masaaki Sato;Kazunori Suzuki;Hiroshi Yamazaki;S. Nakanishi
出生后小脑颗粒细胞的原代培养提供了一个模型系统，可以概括体内发育中的颗粒细胞的许多分子事件。培养的颗粒细胞去极化增加细胞内钙离子，激活钙离子/钙调素依赖的钙调神经磷酸酶(CaN)。这种Ca(2+)信号模拟体内颗粒细胞增殖、迁移和分化的一些信号事件。我们研究了去极化和CaN调节基因在培养的小鼠颗粒细胞中的全基因组表达谱，并讨论了它们与体内发育中的颗粒细胞的基因调控的相关性。颗粒细胞培养在非去极化条件(5 MM KCl2)和去极化条件(25 MMKCl2)下，分别加入和不加入CaN抑制剂FK506。应用基因芯片技术分析去极化与非去极化以及FK506处理与未处理之间的基因表达谱。去极化和FK506处理都影响大量基因的表达水平，其中大多数是重叠的，然而，这两个处理相反地调节着这些基因的表达水平。重要的是，许多FK506反应基因在体内与出生后颗粒细胞中的基因表达同步上调或下调。FK506下调的基因在增殖/迁移前颗粒细胞中高度表达，其中许多基因编码与细胞增殖、迁移和分化有关的细胞成分。相比之下，FK506上调的基因主要在迁移后颗粒细胞中表达，包括许多与突触传递和调节有关的功能分子。本研究表明，CaN信号在颗粒细胞的发育和突触组织中起着关键作用。
Primary culture of postnatal cerebellar granule cells provides a model system that recapitulates many molecular events of developing granule cells in vivo. Depolarization of cultured granule cells increases intracellular Ca(2+) and activates Ca(2+)/calmodulin-dependent calcineurin (CaN) phosphatase. This Ca(2+) signaling mimics some of the signaling events for proliferation, migration, and differentiation of granule cells in vivo. We investigated the genome-wide expression profiles of depolarization- and CaN-regulated genes in cultured mouse granule cells and addressed their relevance to gene regulation in developing granule cells in vivo. Granule cells were cultured under a nondepolarization condition (5 mM KCl) and a depolarization condition (25 mM KCl) with and without the CaN inhibitor FK506. Gene expression profiles between depolarization and nondepolarization and between FK506 treatment and untreatment were analyzed by microarray techniques. Both depolarization and FK506 treatment influence expression levels of a large number of genes, most of which are overlapping, however, are conversely regulated by these two treatments. Importantly, many of the FK506-responsive genes are up- or down-regulated in parallel with gene expression in postnatal granule cells in vivo. The FK506-down-regulated genes are highly expressed in proliferating/premigratory granule cells and many of these genes encode cellular components involved in cell proliferation, migration, and differentiation. In contrast, the FK506-up-regulated genes are predominantly expressed in postmigratory granule cells, including many functional molecules implicated in synaptic transmission and modulation. This investigation demonstrates that the CaN signaling plays a pivotal role in development and synaptic organization of granule cells during the postnatal period.