Organotypic brain slice cultures of adult transgenic P301S mice--a model for tauopathy studies.
Organotypic brain slice cultures of adult transgenic P301S mice--a model for tauopathy studies.
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DOI：
10.1371/journal.pone.0045017
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Singer D
中科院分区：
文献类型：
--
作者：
Mewes A;Franke H;Singer D
器官型脑片培养代表了单细胞培养和完整动物研究之间的极好折衷，以这种方式取代和减少了动物实验的数量。器官型脑片被广泛应用于建立神经元发育和再生模型以及中风、癫痫和阿尔茨海默病(AD)的神经元病理学研究。AD的特征是两种蛋白改变，即tau蛋白过度磷酸化和淀粉样蛋白β过度沉积，两者都导致小胶质细胞和星形胶质细胞的激活。被激活的神经胶质细胞包围的过度磷酸化的tau沉积，称为神经原纤维缠结(NFTs)，在转基因小鼠中进行建模，例如，Tau病模型P301S。在这项研究中，我们探索了由成熟的成年转基因小鼠制成的器官型脑片培养作为研究AD多因素表型的潜在模型系统的好处和局限性。首先，对7到10个月大的转基因P301S小鼠的新生(P1)和成年器官型脑片培养的活力进行了比较，并用乳酸脱氢酶(LDH)和四甲基偶氮唑蓝((3-(4，5-Dimethylthiazol-2-yl)-2，5-diphenyltetrazolium)检测了15天的活力。新生切片显示出持续较高的活力水平，而成体切片培养的活力在培养过程中显著下降。为了提高成体切片的活力，对不同的制备和培养条件进行了试验，结果表明：切片厚度减小，培养温度较低，CO2浓度为5%。此外，我们提供了大量的免疫组织化学特征，分析了神经元、星形胶质细胞和小胶质细胞的形态，并与新生儿组织进行了比较。到目前为止，只有最大年龄为两个月的青春期动物被用来制备器官型脑片。目前的研究提供了证据，从7到10个月大的小鼠的成年器官型脑片培养，在没有转基因修饰的情况下，经历了缓慢的细胞程序性死亡，这是由神经元修复系统的功能障碍引起的。
Organotypic brain slice cultures represent an excellent compromise between single cell cultures and complete animal studies, in this way replacing and reducing the number of animal experiments. Organotypic brain slices are widely applied to model neuronal development and regeneration as well as neuronal pathology concerning stroke, epilepsy and Alzheimer’s disease (AD). AD is characterized by two protein alterations, namely tau hyperphosphorylation and excessive amyloid β deposition, both causing microglia and astrocyte activation. Deposits of hyperphosphorylated tau, called neurofibrillary tangles (NFTs), surrounded by activated glia are modeled in transgenic mice, e.g. the tauopathy model P301S. In this study we explore the benefits and limitations of organotypic brain slice cultures made of mature adult transgenic mice as a potential model system for the multifactorial phenotype of AD. First, neonatal (P1) and adult organotypic brain slice cultures from 7- to 10-month-old transgenic P301S mice have been compared with regard to vitality, which was monitored with the lactate dehydrogenase (LDH)- and the MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays over 15 days. Neonatal slices displayed a constant high vitality level, while the vitality of adult slice cultures decreased significantly upon cultivation. Various preparation and cultivation conditions were tested to augment the vitality of adult slices and improvements were achieved with a reduced slice thickness, a mild hypothermic cultivation temperature and a cultivation CO2 concentration of 5%. Furthermore, we present a substantial immunohistochemical characterization analyzing the morphology of neurons, astrocytes and microglia in comparison to neonatal tissue. Until now only adolescent animals with a maximum age of two months have been used to prepare organotypic brain slices. The current study provides evidence that adult organotypic brain slice cultures from 7- to 10-month-old mice independently of the transgenic modification undergo slow programmed cell death, caused by a dysfunction of the neuronal repair systems.
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影响因子：
6.1
作者：
Braithwaite SP;Schmid RS;He DN;Sung ML;Cho S;Resnick L;Monaghan MM;Hirst WD;Essrich C;Reinhart PH;Lo DC
通讯作者：
Lo DC
DOI：
10.1016/s0006-291x(84)80190-4
发表时间：
1984-01-01
影响因子：
3.1
作者：
GLENNER, GG;WONG, CW
通讯作者：
WONG, CW
影响因子：
3
作者：
Frantseva, MV;Carlen, PL;El-Beheiry, H
通讯作者：
El-Beheiry, H
影响因子：
2.9
作者：
Finley, M;Fairman, D;Cho, SG
通讯作者：
Cho, SG
影响因子：
5.5
作者：
De Simoni, A;Griesinger, CB;Edwards, FA
通讯作者：
Edwards, FA