Hozumi I, Aquino D A, Norton W T
Department of Neurology, Albert Einstein College of Medicine, Bronx, NY 10461.
Brain Res. 1990 Nov 26;534(1-2):291-4. doi: 10.1016/0006-8993(90)90142-x.
We previously reported that glial fibrillary acidic protein (GFAP) levels increased significantly at 3 days after stab wounds, relative to sham-operated controls, reaching a maximum of 200% of control value at 5-7 days. They then fell to near-normal values by 21 days. To determine whether these protein changes correlated with changes in GFAP mRNA we performed Northern blot analyses. Total RNA, isolated from lesioned, sham-operated and intact rat forebrains, was hybridized with 32P-labeled mouse GFAP cDNA and quantified by densitometry. The maximum increase in total RNA content in lesioned animals was only 20% over controls at 12 h. GFAP mRNA levels increased to 2-fold control values at 6 h and reached 5-fold at 12 h. Thereafter they remained at 3.5- to 6-fold until 5 days and then declined to 1.5-fold by 21 days. The rapid increase of GFAP message at 12 h preceded a significant increase in GFAP by 2 days and the decrease of message after 5 days was more precipitate than the slow decrease in GFAP content. Sham-operated animals showed no significant changes in GFAP mRNA, compared to intact controls, during the period 3 h to 14 days postoperation. GFAP mRNA and GFAP in the stab-wound model reached levels similar to those found in the experimental autoimmune encephalomyelitis (EAE) model, but returned to normal much more rapidly.
我们先前报道,相对于假手术对照组,在刺伤后3天,胶质纤维酸性蛋白(GFAP)水平显著升高,在5 - 7天时达到对照值的200%的最大值。然后在21天时降至接近正常的值。为了确定这些蛋白质变化是否与GFAP mRNA的变化相关,我们进行了Northern印迹分析。从损伤的、假手术的和完整的大鼠前脑中分离出的总RNA,与用32P标记的小鼠GFAP cDNA杂交,并通过光密度测定法定量。损伤动物的总RNA含量在12小时时相对于对照的最大增加仅为20%。GFAP mRNA水平在6小时时增加到对照值的2倍,在12小时时达到5倍。此后,它们在5天内保持在3.5至6倍,然后在21天时降至1.5倍。GFAP信息在12小时时的快速增加比GFAP在2天后的显著增加提前,并且在5天后信息的减少比GFAP含量的缓慢减少更迅速。与完整对照组相比,假手术动物在术后3小时至14天期间GFAP mRNA没有显著变化。刺伤模型中的GFAP mRNA和GFAP达到了与实验性自身免疫性脑脊髓炎(EAE)模型中相似的水平,但恢复正常的速度要快得多。