Matsuda S, Ii Y, Desaki J, Yoshimura H, Okumura N, Sakanaka M
Department of Anatomy, Ehime University School of Medicine, Japan.
Neuroscience. 1994 Apr;59(3):651-62. doi: 10.1016/0306-4522(94)90184-8.
The development of basic fibroblast growth factor-like immunoreactivity was investigated in the nuclei, cell bodies and processes of Purkinje cells with attention to basic fibroblast growth factor-containing neuronal input to the deep cerebellar nuclei. Immunoblot analysis with the use of the antisera against basic fibroblast growth factor revealed that crude homogenate of the developing rat cerebellum exhibits a main band with the same molecular weight (18,000 mol. wt) as basic fibroblast growth factor in all the postnasal stages examined. Cerebellar cells were not labeled with the antisera during embryonic life. Under light microscopy, basic fibroblast growth factor-like immunoreactivity was detected initially in cortical cells located close to deep cerebellar fissures of the newborn rat but not in superficial cortical regions. It was difficult to determine whether or not they are Purkinje cells at the fusiform stage. On postnatal day 7, immunoreactive Purkinje cells were identified throughout the cerebellar cortex, and they expressed basic fibroblast growth factor-like immunoreactivity mainly in the apical cytoplasm and proximal dendrites. From postnatal day 14 to postnatal day 28, basic fibroblast growth factor-like immunoreactivity was noted not only throughout the cytoplasm of Purkinje cells but also in the nuclei of the immunopositive cells. Our statistical analysis showed that Purkinje cells with nuclear immunoreaction peaked on postnatal day 21. At these stages, nerve fibers immunoreactive for basic fibroblast growth factor were numerous in the cerebellar medulla and deep cerebellar nuclei. After postnatal day 42, Purkinje cells with intense immunoreactivity in the nuclei showed a marked decrease in number, and immunoreactive structures were distributed in the cerebellum in a fashion similar to that in adult rats. Electron microscopy demonstrated that immunoreactivity was located mainly in the apical cytoplasm of Purkinje cells on postnatal day 7 and throughout the cytoplasm and in the nuclear euchromatin from postnatal day 14 to postnatal day 28, as was expected from light-microscopic observations. Immunoreactivity, even though distributed diffusely in the cytoplasm, was absent from the lumen of endoplasmic reticulum and mitochondria. A small population of Purkinje cell axon terminals forming synapses with the soma and dendrites of deep cerebellar nucleus neurons began to express basic fibroblast growth factor on postnatal day 21. This is much later than the starting age for synaptogenesis between Purkinje cells and deep cerebellar nucleus neurons. The age-dependent changes in the localization of basic fibroblast growth factor within Purkinje cell nucleus, soma and processes suggest a complex transport system of this factor within Purkinje cells during postnatal development.
研究了浦肯野细胞的细胞核、细胞体和突起中碱性成纤维细胞生长因子样免疫反应性的发育情况,并关注含有碱性成纤维细胞生长因子的神经元向小脑深部核团的输入。使用抗碱性成纤维细胞生长因子的抗血清进行免疫印迹分析显示,在所有检查的出生后阶段,发育中小鼠小脑的粗匀浆均显示出一条与碱性成纤维细胞生长因子分子量相同(18,000道尔顿)的主要条带。在胚胎期,小脑细胞未被抗血清标记。在光学显微镜下,最初在新生大鼠小脑深部裂附近的皮质细胞中检测到碱性成纤维细胞生长因子样免疫反应性,而在皮质浅部区域未检测到。在梭形阶段很难确定它们是否为浦肯野细胞。在出生后第7天,在整个小脑皮质中鉴定出免疫反应性浦肯野细胞,它们主要在顶端细胞质和近端树突中表达碱性成纤维细胞生长因子样免疫反应性。从出生后第14天到出生后第28天,不仅在浦肯野细胞的整个细胞质中,而且在免疫阳性细胞的细胞核中都注意到了碱性成纤维细胞生长因子样免疫反应性。我们的统计分析表明,具有核免疫反应的浦肯野细胞在出生后第21天达到峰值。在这些阶段,小脑髓质和小脑深部核团中有许多对碱性成纤维细胞生长因子呈免疫反应性的神经纤维。出生后第42天之后,细胞核中具有强烈免疫反应性的浦肯野细胞数量明显减少,免疫反应性结构在小脑中的分布方式与成年大鼠相似。电子显微镜显示,如光学显微镜观察所预期的那样,出生后第7天免疫反应性主要位于浦肯野细胞的顶端细胞质中,从出生后第14天到出生后第28天则位于整个细胞质和核常染色质中。免疫反应性尽管在细胞质中呈弥漫性分布,但在内质网腔和线粒体中不存在。一小部分与小脑深部核团神经元的胞体和树突形成突触的浦肯野细胞轴突终末在出生后第21天开始表达碱性成纤维细胞生长因子。这比浦肯野细胞与小脑深部核团神经元之间突触形成的起始年龄要晚得多。碱性成纤维细胞生长因子在浦肯野细胞核、细胞体和突起中的定位随年龄的变化表明,在出生后发育过程中,该因子在浦肯野细胞内存在复杂的转运系统。