Molecular insight into the effects of hypothyroidism on the developing cerebellum
Molecular insight into the effects of hypothyroidism on the developing cerebellum
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DOI：
10.1016/j.bbrc.2005.03.099
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发表时间：
2005-05-20
影响因子：
3.1
通讯作者：
Yauk, C
中科院分区：
文献类型：
--
作者：
Dong, HY;Wade, M;Yauk, C
尽管甲状腺激素对正常大脑发育的重要性已得到公认，但人们对这一作用背后的关键分子事件知之甚少。我们通过使用微阵列比较甲状腺功能正常和甲状腺功能减退幼年小鼠小脑的全基因组基因表达模式，研究了甲状腺激素对发育中大脑的作用的分子基础。从受孕后第 13 天开始，持续用含 0.1% 或 0.04% 6-丙基-2-硫氧嘧啶 (PTU) 的饮用水处理怀孕母鼠，直至断奶，产生甲状腺功能减退的幼仔。在出生后第 15 天 (PND) 从载体和 0.1% PTU 处理的幼崽中收集小脑，并使用安捷伦高密度寡核苷酸芯片对这些幼崽的 mRNA 进行微阵列分析。统计分析 (MAANOVA) 显示 2940 个基因存在显着差异表达，其中包括 1357 个上调基因和 1583 个下调基因。进一步分析（结合 MAANOVA 和 ANOVA）确定了 204 个显着改变的基因。甲状腺功能减退症对雄性幼崽基因表达的影响大于雌性幼崽。使用实时 (RT) PCR 分析证实了多个基因 [Syt12（突触结合蛋白 12）、Rcor（REI 沉默转录因子共阻遏物）、Bag3（Bcl 相关 athanogene 3）、p21、细胞周期蛋白 D、Bax（Bcl2 相关 X 蛋白）和 Pcp2（浦肯野细胞蛋白 2）] 的转录变化。暴露于 PTU 的 PND 15 和 PND 60 幼崽小脑中 Bag3 的表达显着改变，表明甲状腺功能减退的大脑发生了永久性的功能改变。使用 HepG2 细胞在体外证实了甲状腺激素对 Rcor 表达的负调节。除了 Rcor 之外，在甲状腺功能减退动物中，编码 REST（REI 沉默转录因子）途径关键成分的其他几个基因的表达也发生了改变。这些结果表明，对该途径的修改可能在导致甲状腺功能减退大脑发育受损方面发挥重要作用。皇冠版权所有 (c) 2005。由 Elsevier Inc. 出版。保留所有权利。
Despite the recognized importance of thyroid hormones for normal brain development, little is known about the critical molecular events underlying this role. We investigated the molecular basis of thyroid hormone action on the developing brain by comparing genome-wide gene expression patterns in the cerebellum between euthyroid and hypothyroid juvenile mice using microarrays. Pregnant dams were treated with 0.1% or 0.04% 6-propyl-2-thiouracil (PTU) in drinking water continuously from day 13 post conception until weaning to produce hypothyroid pups. Cerebella were collected from vehicle and 0.1% PTU treated pups at post-natal day (PND) 15, and mRNA from these was subjected to microarray analysis using Agilent high-density oligonucleotide chips. Statistical analysis (MAANOVA) revealed significant differential expression in 2940 genes including 1357 up- and 1583 down-regulated genes. Further analysis (combined MAANOVA and ANOVA) identified 204 significantly altered genes. Hypothyroidism had a greater effect on gene expression in male than in female pups. Transcriptional changes in several genes [Syt12 (Synaptotagmin 12), Rcor (REI-silencing transcription factor co-repressor), Bag3 (Bcl-associated athanogene 3), p21, cyclin D, Bax (Bcl2-associated X protein), and Pcp2 (Purkinje cell protein 2)] were confirmed using real-time (RT) PCR analysis. Significantly altered expression of Bag3 in cerebella from PND 15 and PND 60 pups exposed to PTU suggests permanent functional alterations in the hypothyroid brain. The thyroid hormone negative regulation of Rcor expression was confirmed in vitro using HepG2 cells. In addition to Rcor, expression of several other genes that code for critical components of the REST (REI-silencing transcription factor) pathway was shown to be altered in hypothyroid animals. These results suggest that modification of this pathway may have a significant role in causing impaired development in the hypothyroid brain. Crown copyright (c) 2005. Published by Elsevier Inc. All rights reserved.