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植物血凝素对神经细胞的体内刺激作用。II. 老年大鼠大脑皮质中总合成率和mRNA合成率的变化

In vivo stimulation of nerve cells by phytohemagglutinin. II. Alterations in the rate of total and mRNA synthesis in the brain cortex of old rats.

作者信息

Semsei I, Zs-Nagy I

出版信息

Arch Gerontol Geriatr. 1983 Dec;2(4):307-13. doi: 10.1016/0167-4943(83)90004-3.

Abstract

Bacto-phytohemagglutinin-P (PHA-P) was administered in form of a single intralumbar injection of 2 mg/100 g body weight dose to 24- to 28-mth-old female CGY rats. The accuracy of the injection technique was checked by adding 2% lidocaine to the injection mixture, which resulted in a transient and symmetric paralysis of the posterior limbs when reaching the cerebrospinal fluid. The total RNA content of the liver and brain cortex were measured, and phenolic extraction of RNA was performed the brain cortex. Poly(A) +RNA (mRNA) was separated from the total RNA of the brain cortex by oligo(dT)-cellulose chromatography. Pulse labeling with tritiated uridine was performed 45 min before killing the animals and the incorporation of the radiolabel was measured in the respective RNA classes and corrected for the nucleotide pool size. The rates of total and mRNA synthesis are expressed in percentages of the young untreated rats and compared to old untreated animals. The effects of PHA-P was studied at 4, 10, 20 and 44 h after its injection. A considerable increase of the total RNA content of the brain cortex was measured during the first 10 h of the experiment followed by a slow decrease. However, the RNA content of the brain cortex was still significantly higher at the end of experiment compared to untreated old rats. The rate of total RNA synthesis increased significantly during the first 10 h and remained constantly high until 44 h. The rate of mRNA synthesis increased to a higher extent than that of the total RNA, and also remained high until 44 h.

摘要

将细菌植物血凝素-P(PHA-P)以2毫克/100克体重的剂量进行单次腰椎注射,给予24至28月龄的雌性CGY大鼠。通过向注射混合物中添加2%利多卡因来检查注射技术的准确性,当到达脑脊液时会导致后肢短暂且对称的麻痹。测量肝脏和大脑皮质的总RNA含量,并对大脑皮质进行RNA的酚提取。通过寡聚(dT)-纤维素色谱法从大脑皮质的总RNA中分离出聚腺苷酸(Poly(A))+RNA(mRNA)。在处死动物前45分钟用氚标记的尿苷进行脉冲标记,并在相应的RNA类别中测量放射性标记的掺入量,并根据核苷酸库大小进行校正。总RNA和mRNA合成速率以未处理的年轻大鼠的百分比表示,并与未处理的老年动物进行比较。在注射PHA-P后4、10、20和44小时研究其效果。在实验的前10小时内测量到大脑皮质的总RNA含量有相当大的增加,随后缓慢下降。然而,与未处理的老年大鼠相比,在实验结束时大脑皮质的RNA含量仍然显著更高。总RNA合成速率在最初10小时内显著增加,并一直保持在较高水平直到44小时。mRNA合成速率的增加幅度高于总RNA,并且也一直保持在较高水平直到44小时。

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