Jacobowitz D M, Kallarakal A T
Laboratory of Clinical Science, NIMH, NIH, Bethesda, MD 20892, USA.
Neurotox Res. 2004;6(4):245-57. doi: 10.1007/BF03033435.
The substantia nigra cells of a normal and Parkinson's disease human brain were obtained by the micropunch procedure and total RNA was isolated. Differential display RT-PCR of the total RNA revealed differentially expressed cDNAs that were identified by sequencing. This resulted in the identification of a panel of known and unknown differentially expressed genes. Complex I (NADH ubiquinone oxidoreductase) and Complex IV (cytochrome oxidase) whose expressions are decreased in Parkinson's disease were reduced in the Parkinson brain. Of the various differentially expressed genes, flotillin-1, also known as reggie-2, was of great interest to us. It is a relatively new protein which is an integral membrane component of lipid rafts and has been implicated in signal transduction pathway events. In situ hybridization histochemical studies with human and rat brain sections revealed the presence of this mRNA in discrete neuronal (and possibly glial) cells of the substantia nigra, locus coeruleus, cortex, hippocampus, hypothalamus, thalamus, motor nuclei, nucleus basalis, raphe nucleus, and other brain regions. Immunohistochemical studies revealed that flotillin-1 is not present in all the regions where the message was found. In the rat brain, the most prominent observation was the revelation of all catecholamine cells (dopamine, norepinephrine, epinephrine) by the flotillin-1 antibody (1:100 dilution). At a more concentrated dilution (1:10) other neuronal cells (e.g., cortex, thalamus, hindbrain) were observed. At both dilutions dense dopaminergic fibers were observed in the rat caudate-putamen, nigrostriatal tract, and substantia nigra. It is significant that there is an increased gene expression of flotillin-1 in the Parkinson substantia nigra/ventral tegmental area. The role of flotillin in these cells is unclear although it is interesting that the reggie-2/flotillin-1 gene was upregulated during retinal axon regeneration in the goldfish visual pathway (Schulte et al., Development 124:577-87, 1997) which suggests that flotillin-1/reggie-2 might play a role in axonal growth from the remaining substantia nigra cells of the Parkinson brain.
通过微量打孔法获取正常人和帕金森病患者大脑的黑质细胞,并分离出总RNA。对总RNA进行差异显示逆转录聚合酶链反应,揭示了通过测序鉴定的差异表达的互补DNA。这导致鉴定出一组已知和未知的差异表达基因。在帕金森病中表达降低的复合体I(NADH泛醌氧化还原酶)和复合体IV(细胞色素氧化酶)在帕金森病大脑中减少。在各种差异表达基因中,浮舰蛋白-1(也称为瑞吉蛋白-2)引起了我们的极大兴趣。它是一种相对较新的蛋白质,是脂筏的一种整合膜成分,并与信号转导途径事件有关。对人和大鼠脑切片进行的原位杂交组织化学研究显示,在黑质、蓝斑、皮质、海马、下丘脑、丘脑、运动核、基底核、中缝核和其他脑区的离散神经元(可能还有神经胶质)细胞中存在这种信使核糖核酸。免疫组织化学研究表明,浮舰蛋白-1并不存在于发现该信使核糖核酸的所有区域。在大鼠大脑中,最显著的观察结果是浮舰蛋白-1抗体(1:100稀释)揭示了所有儿茶酚胺能细胞(多巴胺、去甲肾上腺素、肾上腺素)。在更高浓度的稀释度(1:10)下,观察到了其他神经元细胞(如皮质、丘脑、后脑)。在两种稀释度下,在大鼠尾状核-壳核、黑质纹状体束和黑质中都观察到了密集的多巴胺能纤维。值得注意的是,帕金森病黑质/腹侧被盖区中浮舰蛋白-1的基因表达增加。尽管有趣的是,瑞吉蛋白-2/浮舰蛋白-1基因在金鱼视觉通路的视网膜轴突再生过程中上调(舒尔特等人,《发育》124:577 - 587,1997年),这表明浮舰蛋白-1/瑞吉蛋白-2可能在帕金森病大脑中剩余黑质细胞的轴突生长中发挥作用,但浮舰蛋白在这些细胞中的作用尚不清楚。