Fujita Hirofumi, Aoki Hanako, Ajioka Itsuki, Yamazaki Maya, Abe Manabu, Oh-Nishi Arata, Sakimura Kenji, Sugihara Izumi
Department of Systems Neurophysiology, Tokyo Medical and Dental University Graduate School, Bunkyo-ku, Tokyo, Japan ; Systems Neurobiology Laboratory, The Salk Institute for Biological Studies, La Jolla, California, United States of America ; Department of Otolaryngology-Head and Neck Surgery, The Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America.
Department of Systems Neurophysiology, Tokyo Medical and Dental University Graduate School, Bunkyo-ku, Tokyo, Japan.
PLoS One. 2014 Jan 27;9(1):e86679. doi: 10.1371/journal.pone.0086679. eCollection 2014.
Aldolase C (Aldoc, also known as "zebrin II"), a brain type isozyme of a glycolysis enzyme, is expressed heterogeneously in subpopulations of cerebellar Purkinje cells (PCs) that are arranged longitudinally in a complex striped pattern in the cerebellar cortex, a pattern which is closely related to the topography of input and output axonal projections. Here, we generated knock-in Aldoc-Venus mice in which Aldoc expression is visualized by expression of a fluorescent protein, Venus. Since there was no obvious phenotypes in general brain morphology and in the striped pattern of the cerebellum in mutants, we made detailed observation of Aldoc expression pattern in the nervous system by using Venus expression in Aldoc-Venus heterozygotes. High levels of Venus expression were observed in cerebellar PCs, cartwheel cells in the dorsal cochlear nucleus, sensory epithelium of the inner ear and in all major types of retinal cells, while moderate levels of Venus expression were observed in astrocytes and satellite cells in the dorsal root ganglion. The striped arrangement of PCs that express Venus to different degrees was carefully traced with serial section alignment analysis and mapped on the unfolded scheme of the entire cerebellar cortex to re-identify all individual Aldoc stripes. A longitudinally striped boundary of Aldoc expression was first identified in the mouse flocculus, and was correlated with the climbing fiber projection pattern and expression of another compartmental marker molecule, heat shock protein 25 (HSP25). As in the rat, the cerebellar nuclei were divided into the rostrodorsal negative and the caudoventral positive portions by distinct projections of Aldoc-positive and negative PC axons in the mouse. Identification of the cerebellar Aldoc stripes in this study, as indicated in sample coronal and horizontal sections as well as in sample surface photos of whole-mount preparations, can be referred to in future experiments.
醛缩酶C(Aldoc,也称为“zebrin II”)是一种糖酵解酶的脑型同工酶,在小脑浦肯野细胞(PCs)亚群中呈异质性表达,这些浦肯野细胞在小脑皮质中以复杂的条纹模式纵向排列,该模式与输入和输出轴突投射的拓扑结构密切相关。在此,我们构建了敲入Aldoc-维纳斯小鼠,其中通过荧光蛋白维纳斯的表达来可视化Aldoc的表达。由于突变体的一般脑形态和小脑条纹模式没有明显的表型,我们利用Aldoc-维纳斯杂合子中的维纳斯表达对神经系统中Aldoc的表达模式进行了详细观察。在小脑浦肯野细胞、耳蜗背核的车轮状细胞、内耳感觉上皮以及所有主要类型的视网膜细胞中观察到高水平的维纳斯表达,而在背根神经节的星形胶质细胞和卫星细胞中观察到中等水平的维纳斯表达。通过连续切片对齐分析仔细追踪了不同程度表达维纳斯的浦肯野细胞的条纹排列,并将其映射到整个小脑皮质的展开图上,以重新识别所有单独的Aldoc条纹。在小鼠绒球中首次确定了Aldoc表达的纵向条纹边界,并且该边界与攀缘纤维投射模式以及另一种分区标记分子热休克蛋白25(HSP25)的表达相关。与大鼠一样,在小鼠中,小脑核被Aldoc阳性和阴性浦肯野细胞轴突的不同投射分为吻背侧阴性部分和尾腹侧阳性部分。本研究中小脑Aldoc条纹的识别,如样本冠状和水平切片以及整装标本的样本表面照片所示,可在未来实验中参考。