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大鼠小脑皮质和蜗神经核的单极刷状细胞钙视网膜蛋白呈阳性:一项光镜和电镜免疫细胞化学研究。

The unipolar brush cells of the rat cerebellar cortex and cochlear nucleus are calretinin-positive: a study by light and electron microscopic immunocytochemistry.

作者信息

Floris A, Diño M, Jacobowitz D M, Mugnaini E

机构信息

Laboratory of Neuromorphology, Biobehavioral Sciences Graduate Program, University of Connecticut, Storrs 06269-4154.

出版信息

Anat Embryol (Berl). 1994 Jun;189(6):495-520. doi: 10.1007/BF00186824.

Abstract

Cell class-specific markers are powerful tools for the study of individual neuronal populations. The peculiar unipolar brush cells of the mammalian cerebellar cortex have only recently been definitively identified by means of the Golgi method, and we have explored markers of cerebellar neurons with the purpose of facilitating the analysis of this new cell population and, especially, its distribution and ultrastructural features. By light microscopic immunocytochemistry, we demonstrate that, in the rat, the unipolar brush cells are the cortical neurons that are most densely immunostained with antiserum to calretinin, a recently discovered calcium-binding protein. The unipolar brush cells are highly concentrated in the flocculo-nodular lobe, the ventral uvula and the ventral paraflocculus, occur at relatively high density in the lingula, at moderate-to-low density in other folia of the vermis and in the narrow intermediate cortex, and at low to very low density, with the exception of a few hot spots, in the lateral regions of the cerebellar hemispheres and in the dorsal paraflocculus. Unipolar brush cells are also found in the cochlear nucleus. In addition to the unipolar brush cells, calretinin antibody distinctly stains certain mossy fibers, and weakly to moderately stains other cerebellar elements, such as granule neurons and climbing fibers. In the lobules containing high densities of unipolar brush cells, the granule cell bodies and the parallel fibers are much less immunoreactive, and there are many more densely immunostained mossy fibers than in the lobules, where these cells are rare, which suggests some relationships between these elements. In the cerebellar nuclei, small neurons are densely immunostained, while large neurons are immunonegative. The unipolar brush cells reside nearly exclusively in the granular layer. They are small neurons, intermediate in size between granule cells and Golgi cells, and their features are remarkably similar across all lobules. They usually have a single, relatively thick dendrite of varying length that terminates in a brush-like tip consisting of several short branchlets. Utilizing a pre-embedding protocol, we have identified unipolar brush cells with the electron microscope. The cytoplasm of these cells is partially obscured by the electron dense product of calretinin immunoreaction in all regions of the soma and processes. The cells are often covered with non-synaptic appendages and contain a peculiar cytoplasmic inclusion consisting of ringlet subunits. Other characteristic components are numerous neurofilaments, mitochondria and large, dense-core vesicles. Individual brushes enter one or two glomeruli, where the dendritic branchlets establish an unusually extensive synapse with mossy fiber rosettes.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

细胞类别特异性标记物是研究单个神经元群体的有力工具。哺乳动物小脑皮质中独特的单极刷状细胞直到最近才通过高尔基方法得以明确鉴定,我们探索了小脑神经元的标记物,旨在便于分析这个新的细胞群体,尤其是其分布和超微结构特征。通过光学显微镜免疫细胞化学方法,我们证明,在大鼠中,单极刷状细胞是对钙视网膜蛋白抗血清免疫染色最密集的皮质神经元,钙视网膜蛋白是一种最近发现的钙结合蛋白。单极刷状细胞高度集中在绒球小结叶、腹侧蚓垂和腹侧旁绒球,在舌叶中密度相对较高,在蚓部的其他小叶和狭窄的中间皮质中密度为中到低,在小脑半球外侧区域和背侧旁绒球中密度低至极低,但有一些热点区域除外。单极刷状细胞也存在于蜗神经核中。除了单极刷状细胞外,钙视网膜蛋白抗体还能清晰地标记某些苔藓纤维,并对其他小脑成分,如颗粒神经元和攀缘纤维进行弱阳性至中等强度的染色。在含有高密度单极刷状细胞的小叶中,颗粒细胞体和平行纤维的免疫反应性要低得多,且与这些细胞稀少的小叶相比,免疫染色密集的苔藓纤维更多,这表明这些成分之间存在某种关系。在小脑核中,小神经元免疫染色密集,而大神经元免疫阴性。单极刷状细胞几乎完全位于颗粒层。它们是小神经元,大小介于颗粒细胞和高尔基细胞之间,其特征在所有小叶中都非常相似。它们通常有一条单一的、相对较粗的、长度不等的树突,末端是由几个短分支组成的刷状尖端。利用包埋前方案,我们用电子显微镜鉴定了单极刷状细胞。在细胞体和突起的所有区域,这些细胞的细胞质都被钙视网膜蛋白免疫反应的电子致密产物部分遮盖。这些细胞常被非突触附属物覆盖,并含有由小环亚基组成的特殊细胞质内含物。其他特征性成分包括大量神经丝、线粒体和大的致密核心囊泡。单个刷状结构进入一个或两个小球体,在那里树突分支与苔藓纤维花结形成异常广泛的突触。(摘要截取自400字)

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