Department of Systems Neurophysiology, Tokyo Medical and Dental University, Tokyo, Japan.
Graduate School of Informatics, Kyoto University, Kyoto, Japan.
J Neurosci Res. 2018 Sep;96(9):1476-1489. doi: 10.1002/jnr.24206. Epub 2018 Jan 10.
Regional differences in dendritic architecture can influence connectivity and dendritic signal integration, with possible consequences for neuronal computation. In the cerebellum, analyses of Purkinje cells (PCs), which are functionally critical as they provide the sole output of the cerebellar cortex, have suggested that the cerebellar cortex is not uniform in structure as traditionally assumed. However, the limitations of traditional staining methods and microscopy capabilities have presented difficulties in investigating possible local variations in PC morphology. To address this question, we used male mice expressing green fluorescent protein selectively in PCs. Using Neurolucida 360 with confocal image stacks, we reconstructed dendritic arbors of PCs residing in lobule V (anterior) and lobule IX (posterior) of the vermis. We then analyzed morphologies of individual arbors and the structure of the assembled "jungle," comparing these features across anatomical locations and age groups. Strikingly, we found that in lobule IX, half of the reconstructed PCs had two primary dendrites emanating from their soma, whereas fewer than a quarter showed this characteristic in lobule V. Furthermore, PCs in lobule V showed more efficient spatial occupancy compared to lobule IX, as well as greater packing density and increased arbor overlap in the adult. When analyzing complete ensembles of PC arbors, we also observed "hot spots" of increased dendritic density in lobule V, whereas lobule IX showed a more homogeneous spread of dendrites. These differences suggest that input patterns and/or physiology of PCs could likewise differ along the vermis, with possible implications for cerebellar function.
树突形态的区域性差异会影响连接和树突信号整合,从而可能对神经元计算产生影响。在小脑,对浦肯野细胞(PC)的分析表明,小脑皮质的结构并不像传统假设的那样均匀,因为它们作为小脑皮质的唯一输出具有重要的功能。然而,传统染色方法和显微镜功能的局限性给研究 PC 形态的局部变化带来了困难。为了解决这个问题,我们使用雄性小鼠,在其 PC 中选择性地表达绿色荧光蛋白。利用 Neurolucida 360 和共聚焦图像堆栈,我们重建了位于蚓部 V(前部)和 IX(后部)叶的 PC 的树突分支。然后,我们分析了个体树突分支的形态和组装的“丛林”结构,比较了这些特征在解剖位置和年龄组之间的差异。引人注目的是,我们发现,在 IX 叶中,一半的重建 PC 有两个从其胞体发出的主树突,而在 V 叶中,不到四分之一的 PC 具有这种特征。此外,与 IX 叶相比,V 叶中的 PC 具有更高的空间占有率、更大的堆积密度和更多的成年树突分支重叠。当分析完整的 PC 树突分支集合时,我们还观察到 V 叶中树突密度增加的“热点”,而 IX 叶中树突的分布更为均匀。这些差异表明,PC 的输入模式和/或生理学也可能沿着蚓部不同,这可能对小脑功能有影响。