Human narcolepsy susceptibility candidate genes analyzed by microarray, quantitative RT-PCR and distribution shown by in situ hybridization in mice.

n.t.; not tested, n.e.; no expression, n.c.; no coexpression in perifornical hypocretin area.probe set ID is from aAffymetrix, QRT-PCR probe ID is from Applied Biosystems, and All clones for in situ hybridization are IMAGE clones except where noted.Gene expression was compared between 6 narcolepsy and 8 control postmortem posterior hyptothalami (A and B Genechip). Narcolepsy candidate genes selected by statistical analysis of microarray data are listed at left, quantitative RT-PCR results are in the center, and results of in situ hybridization in mouse hypothalamus are at the right. As only downregulated genes were confirmed by RT-PCR studies, upregulated candidates are not shown. Genes labeled with ��validated�� were confirmed as narcolepsy related genes. Note that IGFBP3 is the only gene validated by QRT-PCR and showing a hypocretin-like distribution pattern. Some candidates showed uniform signal over the whole brain section, potentially indicating ubiquitous expression, but we regarded the staining as background and classified the gene as not expressed if no anatomical variation in staining level was observed.n.t.（未检测，not tested）、n.e.（无表达，no expression）、n.c.（无共表达，no coexpression）于穹窿周促食欲素区（perifornical hypocretin area）。探针组ID来自Affymetrix公司，定量实时荧光PCR（QRT-PCR）探针ID来自应用生物系统公司（Applied Biosystems）；除特别注明外，所有原位杂交（in situ hybridization）所用克隆均为IMAGE克隆（IMAGE clones）。本研究通过基因芯片（A和B芯片）比较了6例发作性睡病患者与8例对照者的死后下丘脑后组织的基因表达差异。通过微阵列数据统计分析筛选出的发作性睡病候选基因列于左侧，定量逆转录PCR（qRT-PCR）结果位于中间，小鼠下丘脑原位杂交结果则在右侧。由于仅下调基因经RT-PCR实验得到验证，因此上调候选基因未在本研究中展示。标注为"validated"的基因已被证实与发作性睡病相关。需注意的是，胰岛素样生长因子结合蛋白3（IGFBP3）是唯一经QRT-PCR验证且呈现促食欲素样分布模式的基因。部分候选基因在全脑切片上呈现均匀信号，这可能提示其为普遍表达，但我们将此类染色信号视为背景，若未观察到染色水平的解剖学差异，则将该基因归类为无表达。