Bicanic Ivana, Hladnik Ana, Petanjek Zdravko
Department of Anatomy and Clinical Anatomy, School of Medicine, University of ZagrebZagreb, Croatia.
Department of Neuroscience, Croatian Institute for Brain Research, School of Medicine, University of ZagrebZagreb, Croatia.
Front Neuroanat. 2017 Apr 28;11:37. doi: 10.3389/fnana.2017.00037. eCollection 2017.
In this study we have provided a detailed quantitative morphological analysis of medium spiny neurons (MSNs) in the mice dorsal striatum and determined the consistency of values among three groups of animals obtained in different set of experiments. Dendritic trees of 162 Golgi Cox (FD Rapid GolgiStain Kit) impregnated MSNs from 15 adult C57BL/6 mice were 3-dimensionally reconstructed using Neurolucida software, and parameters of dendritic morphology have been compared among experimental groups. The parameters of length and branching pattern did not show statistically significant difference and were highly consistent among groups. The average neuronal soma surface was between 160 μm and 180 μm, and the cells had 5-6 primary dendrites with close to 40 segments per neuron. Sholl analysis confirmed regular pattern of dendritic branching. The total length of dendrites was around 2100 μm with the average length of individual branching (intermediate) segment around 22 μm and for the terminal segment around 100 μm. Even though each experimental group underwent the same strictly defined protocol in tissue preparation and Golgi staining, we found inconsistency in dendritic volume and soma surface. These changes could be methodologically influenced during the Golgi procedure, although without affecting the dendritic length and tree complexity. Since the neuronal activity affects the dendritic thickness, it could not be excluded that observed volume inconsistency was related with functional states of neurons prior to animal sacrifice. Comprehensive analyses of tree complexity and dendritic length provided here could serve as an additional tool for understanding morphological variability in the most numerous neuronal population of the striatum. As reference values they could provide basic ground for comparisons with the results obtained in studies that use various models of genetically modified mice in explaining different pathological conditions that involve MSNs.
在本研究中，我们对小鼠背侧纹状体中的中等棘状神经元（MSNs）进行了详细的定量形态分析，并确定了在不同实验组中获得的三组动物之间数值的一致性。使用Neurolucida软件对来自15只成年C57BL/6小鼠的162个经高尔基·考克斯（FD快速高尔基染色试剂盒）浸染的MSNs的树突进行了三维重建，并比较了各实验组之间的树突形态参数。长度和分支模式参数在各实验组之间未显示出统计学上的显著差异，且高度一致。神经元胞体的平均表面积在160μm至180μm之间，细胞有5 - 6个初级树突，每个神经元约有40个节段。Sholl分析证实了树突分支的规则模式。树突的总长度约为2100μm，单个分支（中间）节段的平均长度约为22μm，末端节段约为100μm。尽管每个实验组在组织制备和高尔基染色过程中都遵循了相同的严格定义的方案，但我们发现树突体积和胞体表面积存在不一致。这些变化可能在高尔基染色过程中受到方法学的影响，尽管不影响树突长度和树的复杂性。由于神经元活动会影响树突厚度，因此不能排除观察到的体积不一致与动物处死前神经元的功能状态有关。这里提供的树复杂性和树突长度的综合分析可以作为理解纹状体中数量最多的神经元群体形态变异性的额外工具。作为参考值，它们可以为与使用各种转基因小鼠模型的研究结果进行比较提供基础，以解释涉及MSNs的不同病理状况。
