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大鼠下丘脑核糖体RNA的类固醇激素调节:使用原位杂交和核糖体DNA前体-产物探针的早期检测

Steroid hormone regulation of ribosomal RNA in rat hypothalamus: early detection using in situ hybridization and precursor-product ribosomal DNA probes.

作者信息

Jones K J, Harrington C A, Chikaraishi D M, Pfaff D W

机构信息

Department of Cell Biology and Anatomy, Chicago Medical School, Illinois 60064.

出版信息

J Neurosci. 1990 May;10(5):1513-21. doi: 10.1523/JNEUROSCI.10-05-01513.1990.

Abstract

In the female rat, behavioral and endocrine aspects of reproduction are controlled, in part, by the action of the steroid hormone estradiol on several regions of the brain, including the ventrolateral portion of the ventromedial hypothalamus (VL-VMN) and the arcuate nucleus of the hypothalamus (ARC). Quantitative assessment of the effects of estradiol on the regulation of ribosomal RNA in rat hypothalamus was accomplished in this study by tandem in situ hybridization experiments with 2 ribosomal DNA probes specific to the initial transcript (precursor) or mature, stable (product) rRNA. This novel approach allowed the regulation of RNA processing by steroid hormones to be analyzed in the individual neuron, a particularly important concern in heterogeneous tissue such as the brain. Estradiol was administered subcutaneously to ovariectomized rats for 15 min, 30 min, or 2 hr, or a discontinuous schedule of 2 hr on/7 hr off/2 hr on. Levels of precursor and product rRNA were measured in VL-VMN and ARC neurons using a computerized image-analysis system. Significant increases in the levels of precursor rRNA were observed only in the VL-VMN as early as 30 min after hormone exposure, with a doubling in the amount of precursor rRNA occurring at 2 hr. No changes in product rRNA were observed in either brain region at these early times. These data, in conjunction with our previous findings of increases in product rRNA after longer hormone exposure times, lead us to conclude that rRNA gene transcription is activated in rat hypothalamic neurons within 30 min.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

在雌性大鼠中,生殖行为和内分泌方面部分受甾体激素雌二醇作用于大脑几个区域的调控,这些区域包括腹内侧下丘脑的腹外侧部分(VL-VMN)和下丘脑弓状核(ARC)。本研究通过使用针对初始转录本(前体)或成熟、稳定(产物)rRNA的两种核糖体DNA探针进行串联原位杂交实验,对雌二醇对大鼠下丘脑核糖体RNA调控作用进行了定量评估。这种新方法能够在单个神经元中分析甾体激素对RNA加工的调控,这在诸如大脑这种异质性组织中是一个特别重要的问题。对卵巢切除的大鼠皮下注射雌二醇15分钟、30分钟或2小时,或者采用2小时注射/7小时不注射/2小时注射的间断给药方案。使用计算机图像分析系统测量VL-VMN和ARC神经元中前体和产物rRNA的水平。早在激素暴露后30分钟,仅在VL-VMN中观察到前体rRNA水平显著增加,2小时时前体rRNA量增加一倍。在这些早期时间点,两个脑区的产物rRNA均未观察到变化。这些数据,结合我们之前发现的较长激素暴露时间后产物rRNA增加的结果,使我们得出结论:大鼠下丘脑神经元中的rRNA基因转录在30分钟内被激活。(摘要截短至250字)

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