ONTOGENY OF AROMATASE MESSENGER-RIBONUCLEIC-ACID IN MOUSE-BRAIN - FLUOROMETRICAL QUANTITATION BY POLYMERASE CHAIN-REACTION
ONTOGENY OF AROMATASE MESSENGER-RIBONUCLEIC-ACID IN MOUSE-BRAIN - FLUOROMETRICAL QUANTITATION BY POLYMERASE CHAIN-REACTION
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DOI：
10.1210/en.131.5.2306
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发表时间：
1992-11-01
期刊：
影响因子：
4.8
通讯作者：
YAMADA, K
中科院分区：
文献类型：
--
作者：
HARADA, N;YAMADA, K
建立了一种灵敏的测定各种组织中芳香化酶mRNA的方法。通过在内标RNA的存在下进行逆转录，然后用荧光引物通过聚合酶链反应扩增所得cDNA，对芳香化酶mRNA进行定量。用聚合酶链反应方法检测芳香化酶mRNA在小鼠体内的组织分布。结果表明，芳香化酶mRNA在脑、睾丸和卵巢中表达，但在其他组织中几乎不表达，包括胎盘。在小鼠脑中，芳香化酶主要位于前脑，尤其是间脑。在成年男性和女性间脑中，芳香化酶mRNA水平分别为0.022 +/- 0.004和0.014 +/- 0.003阿托摩尔/μ g总RNA。已知间脑中的芳香化酶参与脑分化和性行为，因此研究了发育期间脑中芳香化酶mRNA水平的变化。芳香化酶mRNA首先在12日龄的胎儿中检测到，并发现在胎儿和新生儿期间迅速增加。其mRNA水平在雄性和雌性大脑中分别在出生后3-4天达到0.068 +/- 0.008和0.059 +/- 0.006阿托摩尔/微克总RNA的最大值，然后逐渐下降到成年水平。这些观察结果是一致的，与以前的数据表明，边缘期的新生儿印迹的性别差异是在出生后一周内，并建议大脑芳香化酶可能是重要的性分化和行为。
A sensitive method was developed for the determination of trace amounts of aromatase mRNA in various tissues. Aromatase mRNA was quantitated by subjecting it to reverse transcription in the presence of an internal standard RNA and then amplifying the resulting cDNA by polymerase chain reaction with a fluorescent primer. The tissue distribution of aromatase mRNA in mice was examined by this polymerase chain reaction method. Results showed that aromatase mRNA is expressed in the brain, testis, and ovary, but scarcely at all in other tissues, including the placenta. In mouse brain, aromatase is mainly located in the forebrain, especially the diencephalon. In adult male and female diencephala, aromatase mRNA levels were 0.022 +/- 0.004 and 0.014 +/- 0.003 attomoles/mug total RNA, respectively. Aromatase in the diencephalon is known to participate in brain differentiation and sexual behavior, so changes in its mRNA levels in the brain during development were examined. Aromatase mRNA was first detected in the 12-day-old fetus and was found to increase rapidly during the fetal and neonatal periods. Its mRNA levels in male and female brains reached maxima of 0.068 +/- 0.008 and 0.059 +/- 0.006 attomoles/mug total RNA, respectively, 3-4 days after birth and then gradually decreased to adult levels. These observations are consistent with previous data indicating that the marginal period of neonatal imprinting of sexual differences is within a week after birth and suggest that brain aromatase may be important in sexual differentiation and behavior.