SYNAPTOPHYSIN GENE-EXPRESSION IN HUMAN BRAIN - A QUANTITATIVE IN-SITU HYBRIDIZATION AND IMMUNOCYTOCHEMICAL STUDY
SYNAPTOPHYSIN GENE-EXPRESSION IN HUMAN BRAIN - A QUANTITATIVE IN-SITU HYBRIDIZATION AND IMMUNOCYTOCHEMICAL STUDY
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DOI：
10.1016/0306-4522(94)90292-5
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发表时间：
1994-04-01
期刊：
影响因子：
3.3
通讯作者：
HARRISON, PJ
中科院分区：
文献类型：
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作者：
EASTWOOD, SL;BURNET, PWJ;HARRISON, PJ
突触素是一种突触前囊泡蛋白。其定量检测已成为突触密度的一种分子标志物。我们利用原位杂交组织化学方法研究了新皮质、海马和小脑中的突触素信使RNA，以确定在人死后的大脑中是否能检测到编码转录本，并研究可能影响其丰度的因素。还对海马中的突触素进行了免疫细胞化学检测。在所有检测的新皮质区域，突触素信使RNA的表达水平是一致的。在各个区域之间以及两个半球的同源区域之间，发现突触素信使RNA的量具有很强的相关性。突触素信使RNA随着年龄的增长和死后间隔时间的延长而减少。突触素免疫反应性也随着年龄以及福尔马林固定时间的延长而降低，但与死后间隔时间无关。突触素免疫反应性与每个海马亚区内部以及投射到该亚区的神经元中信使RNA的丰度相关。在左侧CA4和CA1区域观察到的突触素免疫反应性明显高于右侧。这些数据表明，在人死后的大脑切片中可以可靠地对突触素信使RNA以及突触素本身进行定量检测。它们与其他研究结果一致，即新皮质的突触密度相对均匀且随年龄增长而降低。它们还提示海马突触素表达可能存在不对称性。突触素信使RNA的水平与突触素免疫反应性平行，表明基因表达的变化导致了在衰老和其他情况下观察到的后者的变化。突触素信使RNA的检测拓宽了研究突触蛋白基因产物的方法范围，并可将其用作人类大脑生理和病理过程中突触密度和突触受累的标志物。
Synaptophysin is a presynaptic vesicle protein. Its quantitative detection has become established as a molecular marker of synaptic density. We have studied synaptophysin messenger RNA in the neocortex, hippocampus and cerebellum using in situ hybridization histochemistry to see if the encoding transcript can be detected in post mortem human brain and to investigate factors which might influence its abundance. Synaptophysin was also measured immunocytochemically in the hippocampus. The level of synaptophysin messenger RNA expression was uniform in all neocortical areas examined. Strong correlations were found for the amount of synaptophysin messenger RNA between individual regions and between homologous areas in the two hemispheres. Synaptophysin messenger RNA declined with increasing age and prolonged post mortem interval. Synaptophysin immunoreactivity also reduced with age, as well as with duration of formalin fixation but not post mortem interval. Synaptophysin immunoreactivity correlated with the abundance of the messenger RNA in neurons within, and projecting to, each hippocampal subfield. Significantly greater synaptophysin immunoreactivity was seen in the left than the right CA4 and CA1 regions.These data show that quantitative detection of synaptophysin messenger RNA as well as synaptophysin itself can reliably be carried out in post mortem human brain sections. They are in keeping with other findings that synaptic density is relatively uniform through the neocortex and decreases with age. They also suggest a possible asymmetry of hippocampal synaptophysin expression. The level of synaptophysin messenger RNA paralleled that of synaptophysin immunoreactivity, indicating that changes in gene expression contribute to variations in the latter observed in aging and other situations. Detection of synaptophysin messenger RNA broadens the range of methods by which synaptic protein gene products can be studied and used as markers of synaptic density and synaptic involvement during physiological and pathological processes in human brain.