Suppression subtractive hybridization and microarray identification of estrogen-regulated hypothalamic genes
Suppression subtractive hybridization and microarray identification of estrogen-regulated hypothalamic genes
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DOI：
10.1023/b:nere.0000023606.13670.1d
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发表时间：
2004-06-01
影响因子：
4.4
通讯作者：
Ronnekleiv, OK
中科院分区：
文献类型：
--
作者：
Malyala, A;Pattee, P;Ronnekleiv, OK
性腺类固醇雌激素是一种多效性激素，对多种细胞功能具有多种作用。雌激素的主要目标之一是大脑，类固醇不仅影响神经元的生长、分化和存活，还调节细胞的兴奋性。由于雌激素调节多个重叠的信号通路，因此很难仔细检查类固醇的转录活性。因此，我们仍然缺乏不同基因如何相互作用以及如何受雌激素调节的全局图景。在此，我们报告了使用抑制消减杂交，然后对在雌性生殖周期的负反馈阶段差异表达的数千个基因进行定制微阵列分析。我们发现了许多受雌激素调节的关键转录本，这些转录本有助于改变突触传递，从而改变下丘脑神经元（例如 GABA 神经元）的兴奋性。其中包括 gec-1、GABA(B)R2、PI3 激酶亚基 p55gamma 以及许多含有对质膜靶向至关重要的 pleckstrin 同源结构域的蛋白质。正在进行研究。我们对单个细胞核和单个细胞进行分析。然而，从这种高度敏感的微阵列分析中得出的结论是，雌激素不仅通过新膜蛋白（例如受体和通道）的转录，而且通过改变下游信号分子和参与神经分泌途径的蛋白质的表达来影响下丘脑神经元的神经元可塑性。
The gonadal steroid estrogen is a pleiotropic hormone that has multiple effects on numerous cellular functions. One of estrogen's major targets is the brain, where the steroid not only affects growth, differentiation, and survival of neurons, but also regulates cell excitability. Because estrogen modulates multiple, overlapping signaling pathways, it has been difficult to scrutinize the transcriptional activity of the steroid. Therefore, we still lack a global picture of how different genes interact and are regulated by estrogen. Herein we report the use of suppression subtractive hybridization followed by custom microarray analysis of thousands of genes that are differentially expressed during the negative feedback phase of the female reproductive cycle. We have found a number of key transcripts that are regulated by estrogen and contribute to the alteration in synaptic transmission and hence excitability of hypothalamic neurons (e. g., GABA neurons). These include gec-1, GABA(B)R2, PI3 kinase subunit p55gamma, and a number of proteins containing pleckstrin homology domains that are critical for plasma membrane targeting. Studies are underway to re. ne our analysis to individual nuclei and individual cells. However, what has emerged from this highly sensitive microarray analysis is that estrogen affects neuronal plasticity in hypothalamic neurons not only by transcription of new membrane proteins (e. g., receptors and channels), but also by altering expression of downstream signaling molecules and proteins involved in neurosecretory pathways.