Background

Spine loss is a hallmark of Alzheimer´s and other neurodegenerative diseases, and testing candidate therapeutic drugs needs quantitative analysis of dendritic spine densities. Golgi-Cox impregnation of neurons is a classical method to visualize dendritic spines in diseased brains. Importantly, at early disease stages spine loss occurs locally in the vicinity of amyloid plaques, and concomitant fluorescence labeling of amyloid plaques is required to detect local spine damage.

New Method

Because Golgi-Cox impregnation is done on unsectioned brains, whereas fluorescence staining is performed on sectioned material, the combination is technically challenging. We have now developed a novel combination of Golgi-Cox impregnation with methoxy-X04 fluorescence labeling of plaques that is performed on unsectioned brains.

Results

We used this new combination method to quantify dendritic spine densities in mouse hippocampal CA1 pyramidal neurons. Comparison of neurons from wildtype and APP/PS1 mice revealed local spine loss in the vicinity of amyloid plaques in both male and female APP/PS1 mice.

Comparison with existing method

Golgi-Cox impregnation of neurons combined with methoxy-X04 staining of amyloid plaques is a highly reliable, easy-to-use method for permanent visualization of spines as compared to the technically more sophisticated and less stable fluorescence imaging of spines.

Conclusion

Our novel combination method will be highly useful for testing potential therapeutic drugs in Alzheimer mouse models.

中文翻译：

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高尔基-科克斯（Golgi-Cox）浸渍与淀粉样蛋白斑的荧光染色相结合，揭示了阿尔茨海默病小鼠模型中的局部脊柱丢失。

背景

脊柱丢失是阿尔茨海默氏病和其他神经退行性疾病的标志，测试候选治疗药物需要对树突棘密度进行定量分析。神经元的高尔基-科克斯（Golgi-Cox）浸渍是可视化患病大脑中树突棘的经典方法。重要的是，在疾病的早期阶段，在淀粉样斑块附近局部发生脊柱丢失，并且需要伴随淀粉样斑块的荧光标记来检测局部脊柱损伤。

新方法

因为高尔基-科克斯（Golgi-Cox）浸渍是在未切开的大脑上进行的，而荧光染色是在切开的材料上进行的，所以这种组合在技术上具有挑战性。现在，我们已经开发了一种在未切开的大脑上进行的高尔基体-Cox浸渍与斑块的甲氧基X04荧光标记的新型组合。

结果

我们使用这种新的组合方法来量化小鼠海马CA1锥体神经元中的树突棘密度。来自野生型和APP / PS1小鼠的神经元的比较显示在雄性和雌性APP / PS1小鼠的淀粉样斑块附近存在局部脊柱丢失。

与现有方法的比较

与技术上更为复杂且不稳定的棘突荧光成像相比，神经元的高尔基-科克斯（Golgi-Cox）浸渍与淀粉样蛋白斑的甲氧基X04染色相结合是一种高度可靠，易于使用的方法，可永久显示棘突。

结论

我们的新型组合方法对于在阿尔茨海默病小鼠模型中测试潜在的治疗药物将非常有用。