Endogenous amyloidogenesis in long-term rat hippocampal cell cultures
Endogenous amyloidogenesis in long-term rat hippocampal cell cultures
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DOI：
10.1186/1471-2202-12-38
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发表时间：
2011-05-10
期刊：
影响因子：
2.4
通讯作者：
Booze, Rosemarie M.
中科院分区：
文献类型：
--
作者：
Bertrand, Sarah J.;Aksenova, Marina V.;Booze, Rosemarie M.
背景：长期原代神经元培养是研究神经元衰老相关生化过程的有用工具。现有技术的改进使得有可能在体外长时间观察从啮齿动物脑的不同区域分离的神经细胞的成熟。现有的实验证据表明，细胞老化发生在成熟的，长期的，原代神经元细胞培养。然而，需要详细的研究，在体外神经元的发育，以证明长期的细胞培养为基础的模型，用于调查的生化机制，在体外神经元的发育和senescence.Results的有效性：在目前的研究中，神经元富集的海马细胞培养物被用来分析海马神经元的分化和退化超过两个月的时间段。不同的神经元和星形胶质细胞生物标志物的表达被用来确定海马细胞在不同年龄的长期培养的细胞化学特征。据观察，表达的中间丝巢蛋白是不存在的文化年龄超过21天在体外（DIV），和神经元或星形胶质细胞标记物的表达似乎取代巢蛋白。此外，神经元完整性的形态学评价和Hoescht染色用于评估海马培养物发育和老化过程中的细胞状况。发现A β（1-42）的内源性产生增加，刚果红结合淀粉样蛋白聚集体的积累增加，这与原代培养物中神经元的老化有关。在体外共存的星形胶质细胞和细胞培养的年龄依赖性变性的神经树突网络的形态学的变化类似的功能，在体内脑老化在cellularlevel.Conclusion：总之，本研究表明，长期的主要中枢神经系统培养是一个可行的模型，为研究的基本机制和有效的方法，以减缓神经元衰老的过程。
Background: Long-term primary neuronal cultures are a useful tool for the investigation of biochemical processes associated with neuronal senescence. Improvements in available technology make it possible to observe maturation of neural cells isolated from different regions of the rodent brain over a prolonged period in vitro. Existing experimental evidence suggests that cellular aging occurs in mature, long-term, primary neuronal cell cultures. However, detailed studies of neuronal development in vitro are needed to demonstrate the validity of long-term cell culture-based models for investigation of the biochemical mechanisms of in vitro neuronal development and senescence.Results: In the current study, neuron-enriched hippocampal cell cultures were used to analyze the differentiation and degeneration of hippocampal neurons over a two month time period. The expression of different neuronal and astroglial biomarkers was used to determine the cytochemical characteristics of hippocampal cells in long-term cultures of varying ages. It was observed that the expression of the intermediate filament nestin was absent from cultures older than 21 days in vitro (DIV), and the expression of neuronal or astrocytic markers appeared to replace nestin. Additionally, morphological evaluations of neuronal integrity and Hoescht staining were used to assess the cellular conditions in the process of hippocampal culture development and aging. It was found that there was an increase in endogenous production of A beta(1-42) and an increase in the accumulation of Congo Red-binding amyloidal aggregates associated with the aging of neurons in primary culture. In vitro changes in the morphology of coexisting astrocytes and cell culture age-dependent degeneration of neurodendritic network resemble features of in vivo brain aging at the cellular level.Conclusion: In conclusion, this study suggests that long-term primary CNS culture is a viable model for the study of basic mechanisms and effective methods to decelerate the process of neuronal senescence.