Tandrup Trine
Stereological Research Laboratory, University of Aarhus, DK-8000C Arhus, Denmark.
J Neurocytol. 2004 Mar;33(2):173-92. doi: 10.1023/b:neur.0000030693.91881.53.
For quantitative studies of rat dorsal root ganglion (DRG) in experimental models stereological principles offer a number of different techniques. The application, however, requires knowledge of the anatomy and cytology of the ganglion, considerations of sampling and choosing between the many estimators available. For number and volume estimates in thick glycolmethacrylate sections the optical fractionator and the vertical planar rotator technique in most cases provide sufficient efficiency and are simple to use. Classification of the neurons in the DRG into A- and B-cells is of value in experimental conditions where the two cell types can react differently. Studies on development and subclassification of neuronal DRG cells will gain from application of stereological methods, also. Until now the methods have mainly been applied in studies of axotomy and in a few intoxication models where the time course of cell loss and changes in perikarya volume are important parameters. Further quantitative studies providing better understanding of distribution and expression of neuropeptides, cytokines, apoptotic molecules etc. will provide insight for future therapeutic approaches in neurodegenerative disorders. The more demanding staining techniques require less restrictive embedding media, but unbiased principles are available for almost all the stereological techniques applied to tissue only deformed after sectioning.
在实验模型中,对于大鼠背根神经节(DRG)的定量研究,体视学原理提供了许多不同的技术。然而,这些技术的应用需要了解神经节的解剖学和细胞学知识,考虑抽样问题,并在众多可用的估计方法中进行选择。对于厚甲基丙烯酸乙二醇酯切片中的数量和体积估计,在大多数情况下,光学分割器和垂直平面旋转器技术具有足够的效率且易于使用。在两种细胞类型反应不同的实验条件下,将DRG中的神经元分为A细胞和B细胞具有重要意义。对神经元DRG细胞的发育和亚分类研究也将受益于体视学方法的应用。到目前为止,这些方法主要应用于轴突切断术研究以及少数中毒模型,其中细胞丢失的时间进程和胞体体积变化是重要参数。进一步的定量研究有助于更好地理解神经肽、细胞因子、凋亡分子等的分布和表达,这将为神经退行性疾病的未来治疗方法提供思路。要求更高的染色技术需要限制较少的包埋介质,但几乎所有应用于仅在切片后变形的组织的体视学技术都有非偏倚原理可用。