Effect of haemorrhage on the expression of neurotransmitter-related genes in rat ventrolateral medulla: a quantitative real-time RT-PCR study
Effect of haemorrhage on the expression of neurotransmitter-related genes in rat ventrolateral medulla: a quantitative real-time RT-PCR study
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DOI：
10.1016/s0169-328x(03)00131-1
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发表时间：
2003-05-26
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影响因子：
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通讯作者：
Pilowsky, PM
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作者：
Li, Q;Goodchild, AK;Pilowsky, PM
延髓腹外侧区（VLM）有三个功能明确的区域，其中含有合成儿茶酚胺的神经元（头侧C1，尾侧C1和A1区）。许多神经信使可以改变VLM中的心血管功能。本研究的目的是，首先验证实时RT-PCR SYBR绿色测定用于定量VLM中小位点特异性神经元群体中神经信使mRNA表达水平的实用性，其次比较肾上腺素合成酶苯乙醇胺-N-甲基转移酶（PNMT）的基础mRNA水平，神经肽Y（NPY）和前脑啡肽原（ENK）在三个区域和第三，以检查出血对这三个基因的表达的影响。大鼠麻醉与戊巴比妥钠，并分为三组：灌注，假手术和出血。对出血组进行了15%的出血。结果发现，所有三个基因的mRNA表达水平存在区域差异：一般来说，从吻部到尾侧区域的VLM减少。15%的出血显着诱导PNMT在吻侧C1区和NPY在尾侧C1和A1区的表达，但没有影响ENK在任何网站上，这三个基因在VLM的表达的差异调节。我们的研究结果还表明，实时RT-PCR是一种敏感和准确的定量研究神经递质基因表达的限制性脑区域的方法。皇冠版权所有（C）2003年出版的爱思唯尔科学B. V.保留所有权利。
The ventrolateral medulla (VLM) has three functionally defined regions that contain catecholamine-synthesising neurons (rostral Cl, caudal C1 and A1 regions). Many neuromessengers can alter cardiovascular functions in the VLM. The aims of this study were, first to validate the utility of real-time RT-PCR SYBR Green assay for quantitation of mRNA expression levels of neuromessengers in small site-specific neuronal populations in the VLM, and second to compare the basal mRNA levels of the adrenaline-synthesizing enzyme phenylethanolamine-N-methyltransferase (PNMT), neuropeptide Y (NPY) and preproenkephalin (ENK) in the three regions and third to examine the effects of haemorrhage on the expression of these three genes. Rats were anaesthetised with sodium pentobarbital and divided into three groups: perfused, sham-operated and haemorrhaged. A 15% haemorrhage was carried out on the haemorrhaged group. It was found that there are regional differences in the level of mRNA expression for all the three genes: with, in general, decreases from the rostral to caudal regions of VLM. A 15% haemorrhage significantly induced expression of PNMT in the rostral C1 region and NPY in the caudal C1 and A1 regions but had no effect on ENK at any sites, suggesting a differential regulation on the expression of these three genes in the VLM. Our results also demonstrate that real-time RT-PCR is a sensitive and accurate method for quantitative studies on neurotransmitter gene expressions in restricted brain regions. Crown Copyright (C) 2003 Published by Elsevier Science B.V. All rights reserved.