Neuronal differentiation is accompanied by increased levels of SNAP-25 protein in fetal rat primary cortical neurons - Implications in neuronal plasticity and Alzheimer's disease
Neuronal differentiation is accompanied by increased levels of SNAP-25 protein in fetal rat primary cortical neurons - Implications in neuronal plasticity and Alzheimer's disease
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DOI：
10.1196/annals.1377.001
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发表时间：
2006-01-01
期刊：
影响因子：
--
通讯作者：
Lahiri, Debomoy K.
中科院分区：
文献类型：
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作者：
Bailey, Jason A.;Lahiri, Debomoy K.
阿尔茨海默病（AD）伴随着进行性记忆丧失和一般认知能力下降。神经病理学特征包括淀粉样斑块形成、神经纤维缠结以及神经元和突触损失。为了改善AD的研究模型，我们对原代大鼠神经元细胞培养模型的几个方面进行了表征，这些方面与神经胞吐、神经元可塑性和淀粉样变性的研究直接相关。我们已经试图分离和评估的神经元富集的原代皮层细胞培养的形态学和分子技术，这些培养的细胞培养的进展，为18天的文化，以确定在这些细胞中的操作和数据收集的最佳时间框架。我们在第15天左右观察到最大神经元分化，其在第18天下降，因此，在这些培养物中操作的关键窗口似乎在第12-15天之间。神经炎长度和密度的显著增加在非常早的时间点是明显的，并在第15天达到峰值。在形态学评估的同时，我们测量了一组选定的神经元重要蛋白质的水平，包括细胞骨架和突触蛋白。细胞相关APP和突触素水平的变化不像SNAP-25或分泌型APP那样显著。值得注意的是，这种增加的分化伴随着SNAP-25蛋白水平的显著增加，其在第15天达到峰值，此后下降。这些结果表明，这个神经元群体包含可量化的突触前末梢。因此，这种神经元模型的建立和分子表征将在神经科学研究中具有许多意义，包括突触分化和神经元可塑性以及用于评估疾病过程中的药物干预。
Alzheimer's disease (AD) is accompanied by progressive memory loss and decline in general cognitive abilities. Neuropathological hallmarks include amyloid plaque formation, neurofibrillary tangles, and neuronal and synaptic loss. To improve research models in AD, we have characterized several aspects of the primary rat neuronal cell culture model, which relate directly to the study of neuroexocytosis, neuronal plasticity, and amyloidosis. We have attempted to isolate and assess the neuronal enrichment of a primary cortical cell culture by morphological and molecular techniques, following progress of these cultured cells for up to 18 days in culture to identify the optimum timeframe for the manipulation and data collection in these cells. We observed maximum neuronal differentiation around day 15, which was in decline by day 18, thus, the critical window for manipulations in these cultures seems to be between days 12-15. The substantial increase in neuritic length and density was apparent at very early time points and peaked at day 15. In parallel to morphological assessments, we measured levels of a group of selected neuronally important proteins, including cytoskeletal and synaptic proteins. Change in the levels of cell-associated APP and synaptophysin were not as dramatic as that of SNAP-25 or secreted APP. Notably, this increased differentiation is accompanied by a dramatic increase in levels of SNAP-25 protein, which peaked at day 15 and declined thereafter. These results suggest that this neuronal population contains quantifiable presynaptic terminals. Thus, the establishment and molecular characterization of this neuronal model would have many implications in neuroscience research including synaptic differentiation and neuronal plasticity and for the evaluation of pharmacological interventions in the disease process.