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帕金森病患者肾上腺中β-神经生长因子(β-NGF)mRNA的表达

Beta-nerve growth factor (beta-NGF) mRNA expression in the parkinsonian adrenal gland.

作者信息

Silani V, Pizzuti A, Falini A, Borsani G, Rugarli E I, Melo C A, Sidoli A, Villani F, Baralle F, Scarlato G

机构信息

Dino Ferrari Center, Institute of Neurology, University of Milan Medical School, Italy.

出版信息

Exp Neurol. 1991 Aug;113(2):166-70. doi: 10.1016/0014-4886(91)90172-9.

Abstract

The adrenal gland is a well-demonstrated source for different neurotrophic factors. The presence of the beta-nerve growth factor (beta-NGF) mRNA in the adrenal tissue used for grafting in a Parkinsonian patient is reported here. Adrenal samples were obtained on the day of implantation, and a specific cDNA was synthesized after the extraction of total RNA using a synthetic oligonucleotide as a reverse transcription primer. A 168-bp portion of the cDNA was amplified using two other oligonucleotides as Taq polymerase primers in a polymerase chain reaction. Thirty-two cycles of amplification were performed. The amplification products were identified by agarose gel electrophoresis and Southern blot analysis as a single DNA band hybridizing with a third beta-NGF specific oligonucleotide. The identity of the fragment was confirmed by DNA sequencing. Quantitative analysis demonstrated a beta-NGF mRNA concentration exceeding 5 fg/micrograms of total adrenal RNA. These findings add NGF to the other neurotrophic factors produced by the gland (i.e., basic fibroblast growth factor) and demonstrate the retained functional capacity of the Parkinsonian adrenal to express the beta-NGF mRNA. All these data may assume relevant meaning for neurotransplantation research.

摘要

肾上腺是不同神经营养因子的一个已得到充分证实的来源。本文报道了用于帕金森病患者移植的肾上腺组织中β-神经生长因子(β-NGF)mRNA的存在情况。在植入当天获取肾上腺样本,使用合成寡核苷酸作为逆转录引物提取总RNA后合成特异性cDNA。在聚合酶链反应中,使用另外两种寡核苷酸作为Taq聚合酶引物扩增cDNA的168 bp片段。进行了32个循环的扩增。扩增产物通过琼脂糖凝胶电泳和Southern印迹分析鉴定为与第三种β-NGF特异性寡核苷酸杂交的单一DNA条带。通过DNA测序确认了该片段的身份。定量分析表明,β-NGF mRNA浓度超过每微克肾上腺总RNA 5 fg。这些发现使NGF加入到该腺体产生的其他神经营养因子(即碱性成纤维细胞生长因子)之列,并证明帕金森病患者的肾上腺保留了表达β-NGF mRNA的功能能力。所有这些数据对于神经移植研究可能具有重要意义。

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