Estrogen receptor-dependent genomic responses in the uterus mirror the biphasic physiological response to estrogen
Estrogen receptor-dependent genomic responses in the uterus mirror the biphasic physiological response to estrogen
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DOI：
10.1210/me.2003-0146
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发表时间：
2003-10-01
影响因子：
--
通讯作者：
Korach, KS
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文献类型：
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作者：
Hewitt, SC;Deroo, BJ;Korach, KS
啮齿动物子宫对急性雌激素(E)剂量的生理反应可分为早期事件和晚期事件。早期反应的例子包括分别注射E后2小时和6小时的RNA转录增加、充血和水分吸收，而后期反应包括E后10小时和16小时开始的DNA合成周期和上皮细胞有丝分裂。雌激素受体(ER)基因敲除(ERKO)小鼠的发展，结合微阵列技术，使我们能够设计一种基因组方法来研究啮齿动物生殖道对E的急性反应，以确定早期和晚期的生物反应是否与早期和晚期反应的单组基因或不同组基因的调节改变有关。取去卵巢小鼠的子宫RNA，分别用溶剂或雌二醇处理2 h(早期)或24 h(晚期)。样本也是从同样处理的缺乏ERAlpha(AlphaERKO)或ERbeta(BetaERKO)的小鼠身上制备的，以确定ER在子宫反应中的相对贡献。对8700个小鼠基因表达的芯片分析表明，在早期或晚期受E正向和负向调控的基因簇以及在这两个时间段都受到调控的基因簇。BetaERKO样品的早期和晚期反应与WT样品的反应没有区别，而AlphaERKO样品对E的响应在基因表达上几乎没有变化，表明ERpha在基因组反应中起主导作用。进一步的研究表明，在中间时间点(6h、12h)的样本中，基因组反应落在早期或晚期的簇内，而不是表现出在中间时期调节的独特簇。这种基因组方法的使用说明了生理反应是如何反映在基因组模式中的。此外，识别子宫中受E调控的功能基因家族，结合利用基因改变的实验动物模型，有助于揭示和定义E作用的新机制。
The physiological responses of the rodent uterus to acute estrogen (E) dosing can be divided into early and late events. Examples of early responses include increased RNA transcription, hyperemia, and water imbibition 2 and 6 h following E administration respectively, whereas later responses include cycles of DNA synthesis and mitosis of epithelial cells beginning 10 and 16 h after E. The development of estrogen receptor (ER) knockout (ERKO) mice, combined with microarray technology, has allowed us to design a genomic approach to study the acute response of the rodent reproductive tract to E. To determine whether early and late biological responses are correlated with altered regulation of a single set of genes or distinct sets of genes characteristic of early and late responses, uterine RNA was obtained from ovariectomized mice that were treated with vehicle or with estradiol for 2 h (early) or 24 h (late). Samples were also prepared from identically treated mice that lacked either ERalpha (alphaERKO) or ERbeta (betaERKO) to address the relative contributions of the ERs in the uterine responses. Microarray analysis of the relative expression of 8700 mouse cDNAs indicated distinct clusters of genes that were regulated both positively and negatively by E in the early or late phases as well as clusters of genes regulated at both times. Both early and late responses by the betaERKO samples were indistinguishable from those of WT samples, whereas the alphaERKO showed little change in gene expression in response to E, indicating the predominant role for ERalpha in the genomic response. Further studies indicated that the genomic responses in samples from intermediate time points (6 h, 12 h) fall within the early or late clusters, rather than showing unique clusters regulated in the intermediary period. The use of this genomic approach has illustrated how physiological responses are reflected in genomic patterns. Furthermore, the identification of functional gene families that are regulated by E in the uterus combined with the utilization of genetically altered experimental animal models can help to uncover and define novel mechanisms of E action.