Dournaud P, Cervera-Pierot P, Hirsch E, Javoy-Agid F, Kordon C, Agid Y, Epelbaum J
INSERM U 159, Centre Paul Broca, Paris, France.
Neuroscience. 1994 Aug;61(4):755-64. doi: 10.1016/0306-4522(94)90399-9.
The level of expression of somatostatin messenger RNA-containing neurons in human brain was visualized and quantified by in situ hybridization with a 35S-labelled oligonucleotide complementary to amino acids 96-111 of the preprosomatostatin complementary DNA sequence. The analysis was carried out in the frontal and parahippocampal cortices and hippocampus of six age- and post mortem delay-matched Alzheimer's disease and control brains. By northern blot analysis, in frontal cortex samples, 18S rRNA degradation was identical in control and Alzheimer brains and somatostatin messenger RNAs migrated as a single band of 1 kb. By in situ hybridization, specificity was demonstrated by abolition of the signal using either an excess of unlabelled antisense probe or using a labelled sense probe. Somatostatin messenger RNA-containing neurons displayed a similar regional and subregional distribution in control subjects and patients with Alzheimer's disease, being more abundant in the frontal cortex, followed by the hippocampus and the parahippocampal cortex. An overall reduction of labelled cell density was observed in patients with Alzheimer's disease (frontal cortex gray matter:--41%; white matter:--66%; hippocampus:--44%; parahippocampal cortex white matter:--40%). Due to a great variation between brains, this decrease only reached significance in the parahippocampal cortex (-59%, P < 0.05). A significantly lower level of expression of somatostatin messenger RNA per somatostatinergic cell was observed in the hippocampus of Alzheimer's disease patients (-47%, P < 0.05), but not in frontal cortex gray (-17%) and white (-36%) matter and parahippocampal cortex gray (-42%) and white (-29%) matter. These data are in accordance with the distribution of somatostatin cells as visualized by immunohistochemistry in human brain. They indicate that the ability of cortical cells to express somatostatin messenger RNA is partially preserved in Alzheimer disease brains and that the decrease in the amount of somatostatin messenger RNA per cell is restricted to the hippocampal formation.
采用与前促生长抑素互补DNA序列中第96 - 111位氨基酸互补的35S标记寡核苷酸进行原位杂交，对人脑中含生长抑素信使核糖核酸的神经元的表达水平进行了可视化和定量分析。分析在6例年龄和死后延迟相匹配的阿尔茨海默病患者及对照者的额叶、海马旁回皮质和海马中进行。通过Northern印迹分析，在额叶皮质样本中，对照者和阿尔茨海默病患者的18S核糖体RNA降解情况相同，生长抑素信使核糖核酸迁移为一条1 kb的单带。通过原位杂交，使用过量未标记的反义探针或标记的正义探针消除信号，证明了特异性。含生长抑素信使核糖核酸的神经元在对照者和阿尔茨海默病患者中显示出相似的区域和亚区域分布，在额叶皮质中含量更高，其次是海马和海马旁回皮质。在阿尔茨海默病患者中观察到标记细胞密度总体降低（额叶皮质灰质：-41%；白质：-66%；海马：-44%；海马旁回皮质白质：-40%）。由于不同脑之间差异很大，这种减少仅在海马旁回皮质达到显著水平（-59%，P < 0.05）。在阿尔茨海默病患者的海马中观察到每个生长抑素能细胞的生长抑素信使核糖核酸表达水平显著降低（-47%，P < 0.05），但在额叶皮质灰质（-17%）、白质（-36%）以及海马旁回皮质灰质（-42%）和白质（-29%）中未观察到。这些数据与通过免疫组织化学在人脑中观察到的生长抑素细胞分布一致。它们表明，在阿尔茨海默病脑中，皮质细胞表达生长抑素信使核糖核酸的能力部分保留，且每个细胞中生长抑素信使核糖核酸量的减少仅限于海马结构。
