血清冲击诱导哺乳动物组织培养细胞中昼夜基因表达
A Balsalobre1, F Damiola, U Schibler
1Département de Biologie Moléculaire, Sciences II, Université de Genève, Switzerland.
Cell
|July 11, 1998
概括
用血清治疗的培养细胞表现出昼夜基因表达,反映出体内节律. 这种血清冲击方法为研究昼夜钟基因调节提供了一种新方法.
科学领域:
- 分子生物学分子生物学
- 时间生物学 时间生物学
- 细胞生物学 细胞生物学
背景情况:
- 昼夜节律是内生生物过程,在24小时周期内调节生理功能.
- 基因转录是昼夜节律的关键组成部分,许多基因表现出节奏表达模式.
- 了解围绕昼夜基因表达的分子机制对于破译生物定时至关重要.
研究的目的:
- 研究培养细胞的血清治疗是否可以诱导昼夜基因表达.
- 将培养细胞中的基因表达时间模式与体内观察到的基因表达模式进行比较.
- 探索血清冲击的潜力,作为一种研究昼夜节律的工具.
主要方法:
- 培养的老鼠-1纤维细胞和H35肝瘤细胞被高度的血清治疗.
- 特定基因的mRNA水平,包括rper1,rper2,Rev-Erbα,DBP和TEF,随着时间的推移被量化.
- 在培养细胞中分析了周期长度和振荡次数.
- 在血清休克后,对c-fos和rper表达进行了监测.
主要成果:
- 血清治疗诱导了培养细胞中多个基因的昼夜表达,包括Drosophila时期基因 (rper1和rper2) 的老鼠同类物和转录因子Rev-Erb alpha,DBP和TEF.
- 鼠1型纤维细胞显示了多达三次连续的每日振荡,平均时间为22.5小时.
- 在培养细胞中mRNA积累周期的时间序列与体内观察结果一致.
- 血清冲击暂时刺激了c-fos和rper表达,模仿光诱导的即时早期基因表达.
结论:
- 血清治疗在诱导培养细胞中昼夜基因表达方面是有效的.
- 培养细胞为研究昼夜节律的分子机制提供了有价值的模型.
- 血清冲击方法提供了一种新的方法来研究体外昼夜钟基因调节,可能模仿光感应的反应.
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