Lorent K, Overbergh L, Moechars D, De Strooper B, Van Leuven F, Van den Berghe H
Center for Human Genetics, Katholieke Universiteit van Leuven, Belgium.
Neuroscience. 1995 Apr;65(4):1009-25. doi: 10.1016/0306-4522(94)00555-j.
We have analysed by northern blotting and by in situ hybridization the expression patterns of eight different genes during the second half of mouse embryonic development and in adult mouse brain: we compared the messenger RNA levels of amyloid precursor protein and of the two amyloid precursor protein-like proteins 1 and 2 and we have analysed expression of apolipoprotein E and of its main receptor in brain, the alpha-2-macroglobulin/low density lipoprotein receptor-related protein and three other ligands: the proteinase inhibitor alpha-2-macroglobulin, the modifying enzyme lipoprotein lipase and the 44,000 molecular weight heparin binding protein, a ligand of unknown function. During embryogenesis the temporal expression pattern differs considerably for the three members of the amyloid precursor proteins. Total embryo messenger RNA levels of amyloid precursor protein and amyloid precursor protein-like protein 2 increased progressively, while amyloid precursor protein-like protein 1 messenger RNA showed a burst of synthesis between days 10 and 13 post-coitum. Significantly, expression of the alpha-2-macroglobulin/low density lipoprotein receptor-related protein and of its associated protein, the 44,000 molecular weight heparin binding protein, exhibited their most important increase very similar to that of amyloid precursor protein-like protein 1, between days 10 and 13 post-coitum. Apolipoprotein E, lipoprotein lipase and alpha-2-macroglobulin messenger RNA levels in total embryos increased progressively, beginning most pronounced at days 13, 15 and 17, respectively. In mouse embryos, in situ hybridization established amyloid precursor protein, amyloid precursor protein-like protein 2 and alpha-2-macroglobulin/low density lipoprotein receptor-related protein messenger RNA to be expressed in most organs, with the notable exception of the liver, while expression of the other studied proteins was much more restricted. Among adult mouse tissues, the genes investigated were expressed very prominently in brain, except for lipoprotein lipase and for the complete absence of alpha-2-macroglobulin messenger RNA. In adult mouse brain, the cortex and hippocampus exhibited strong signals for most genes analysed. Exceptions are lipoprotein lipase and apolipoprotein E messenger RNAs, and the absent alpha-2-macroglobulin messenger RNA. Several interesting features, similarities as well as differences, between brain tissue sections hybridized with probes for amyloid precursor protein, amyloid precursor protein-like proteins 1 and 2 and between alpha-2-macroglobulin/low density lipoprotein receptor-related protein and heparin binding protein-44 were observed and are described. The results are further discussed in view of the known or anticipated physiological functions of the proteins examined and of their possible role in the etiology of Alzheimer's disease.
我们通过Northern印迹法和原位杂交技术分析了8种不同基因在小鼠胚胎发育后期及成年小鼠大脑中的表达模式:我们比较了淀粉样前体蛋白、两种类淀粉样前体蛋白1和2的信使RNA水平,并分析了载脂蛋白E及其在大脑中的主要受体、α-2-巨球蛋白/低密度脂蛋白受体相关蛋白以及其他三种配体的表达情况:蛋白酶抑制剂α-2-巨球蛋白、修饰酶脂蛋白脂肪酶以及分子量为44,000的肝素结合蛋白,后者是一种功能未知的配体。在胚胎发生过程中,淀粉样前体蛋白的三个成员的时间表达模式差异很大。淀粉样前体蛋白和类淀粉样前体蛋白2的总胚胎信使RNA水平逐渐升高,而类淀粉样前体蛋白1的信使RNA在交配后第10天至13天之间出现合成高峰。值得注意的是,α-2-巨球蛋白/低密度脂蛋白受体相关蛋白及其相关蛋白——分子量为44,000的肝素结合蛋白的表达,在交配后第10天至13天之间出现了与类淀粉样前体蛋白1非常相似的最重要的增加。总胚胎中的载脂蛋白E、脂蛋白脂肪酶和α-2-巨球蛋白信使RNA水平逐渐升高,分别在第13天、15天和17天开始最为明显。在小鼠胚胎中,原位杂交显示淀粉样前体蛋白、类淀粉样前体蛋白2和α-2-巨球蛋白/低密度脂蛋白受体相关蛋白的信使RNA在大多数器官中表达,但肝脏除外,而其他研究蛋白的表达则更为局限。在成年小鼠组织中,所研究的基因在大脑中表达非常突出,但脂蛋白脂肪酶和完全不存在α-2-巨球蛋白信使RNA的情况除外。在成年小鼠大脑中,皮质和海马体对大多数分析的基因显示出强烈信号。例外的是脂蛋白脂肪酶和载脂蛋白E的信使RNA,以及不存在的α-2-巨球蛋白信使RNA。观察并描述了用淀粉样前体蛋白、类淀粉样前体蛋白1和2的探针杂交的脑组织切片之间,以及α-2-巨球蛋白/低密度脂蛋白受体相关蛋白和肝素结合蛋白-44之间的几个有趣特征,包括相似之处和不同之处。鉴于所检测蛋白质的已知或预期生理功能及其在阿尔茨海默病病因学中的可能作用,对结果进行了进一步讨论。