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通过逆转录聚合酶链反应分析色氨酸羟化酶mRNA:去甲肾上腺素对新生大鼠松果体影响的初步报告

Tryptophan hydroxylase mRNAs analysis by RT-PCR: preliminary report on the effect of noradrenaline in the neonatal rat pineal gland.

作者信息

Besançon R, Reboul A, Claustrat B, Jouvet A, Belin M F, Fèvre-Montange M

机构信息

INSERM U433, Laboratoire de Neurobiologie expérimentale et Physiopathologie, Faculté de Médecine R.T.H. Laënnec, Lyon, France.

出版信息

J Neurosci Res. 1997 Sep 15;49(6):750-8. doi: 10.1002/(SICI)1097-4547(19970915)49:6<750::AID-JNR9>3.0.CO;2-6.

Abstract

The levels of mRNAs coding for tryptophan hydroxylase (TPOH), the first enzyme in melatonin synthesis, have been investigated by quantitative reverse transcription of RNA, followed by polymerase chain reaction (RT-PCR), after stimulation of neonatal pineal organ cultures with Noradrenaline (NA). TPOH mRNAs were specifically amplified from various adult tissues, namely the pineal gland, raphe, retina, and kidney, but not the lung. PCR signals for TPOH were detected in the neonatal pineal gland in the absence of stimulation. Stimulation of neonatal pineal organ culture with 0.1 microM NA resulted in a significant increase (x2.5) in expression of TPOH mRNAs, whereas higher doses (1 and 10 microM) had no effect. All concentrations of NA enhanced melatonin secretion. Our results suggest that the level of TPOH mRNAs can be controlled by NA and that this effect might be implicated in the gene level regulation of the daily enzyme rhythm in the rat pineal gland.

摘要

在用去甲肾上腺素(NA)刺激新生大鼠松果体器官培养物后,通过RNA定量逆转录,随后进行聚合酶链反应(RT-PCR),研究了褪黑素合成的第一种酶——色氨酸羟化酶(TPOH)的mRNA水平。从各种成年组织,即松果体、中缝核、视网膜和肾脏中特异性扩增出了TPOH的mRNA,但在肺中未扩增出。在未刺激的新生松果体中检测到了TPOH的PCR信号。用0.1微摩尔/升的NA刺激新生松果体器官培养物,导致TPOH mRNA的表达显著增加(增加2.5倍),而更高剂量(1和10微摩尔/升)则没有效果。所有浓度的NA都增强了褪黑素的分泌。我们的结果表明,TPOH mRNA的水平可由NA控制,并且这种作用可能与大鼠松果体中每日酶节律的基因水平调节有关。

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