Department of Nutrition, Faculty of Medical and Health Sciences, Tsukuba International University, Tsuchiura, Ibaraki, Japan.
Anat Rec (Hoboken). 2021 Feb;304(2):413-424. doi: 10.1002/ar.24420. Epub 2020 Jun 23.
We immunohistochemically characterized postnatal changes in cerebellar cortical cytoarchitectures in ferrets using markers for cerebellar cortical neurons and glial cells. Although 10 lobules of the vermis were already observed on postnatal day (PD) 4, Purkinje cells were still arrayed into two to three layers. Purkinje cells were aligned in a monolayer by PD 10 and formed mature shapes on PD 42 by developing their dendritic arbors. Parvalbumin immunostaining revealed relatively slower maturation of Purkinje cells in the Lobule X cortex than in other lobules. Basket and stellate cells emerged in the molecular layer on PDs 21 and 42, respectively. Rosette-like arranged glutamate decarboxylase 65 and 67-positive puncta were observed in the inner granular layer (IGL) on PD 21. Proliferating cell nuclear antigen immunostaining appeared in the outer zone of the external granular layer (EGL) containing progenitors of granular neurons on PDs 4-21. Bergmann glial processes extending vertically through the molecular layer and EGL were visible with GFAP immunostaining on PD 10 and thereafter. Their somata, aligned in the Purkinje cell layer, showed immunopositivity to Sox2 already on PD 4 and subsequently to S100 protein on PD 10. Sox2-positive cells were found sparsely in the IGL. Few of them were NeuN positive on PD 90, predicting the possibility of adult neurogenesis. These immunohistochemical results revealed that ferrets underwent cerebellar cortical histogenesis during their postnatal life in sequences. Relatively slow development or maturation of the ferret cerebellum was revealed by the timing of the monolayer alignment and morphological maturation of Purkinje cells.
我们使用标记物对雪貂小脑皮质神经元和神经胶质细胞进行了免疫组织化学分析,以研究其出生后的皮质结构变化。尽管在出生后第 4 天(postnatal day,PD)就已经观察到 10 个小脑蚓部小叶,但浦肯野细胞仍排列成 2-3 层。到 PD10 时,浦肯野细胞排列成单层,到 PD42 时,通过发育其树突形成成熟的形状。巴德蛋白免疫染色显示,在 X 小叶的浦肯野细胞比其他小叶成熟得相对较慢。篮状细胞和星状细胞分别在 PD21 和 PD42 时出现在分子层。在 PD21 时,在内颗粒层(inner granular layer,IGL)中观察到呈玫瑰花结样排列的谷氨酸脱羧酶 65 和 67 阳性点状结构。增殖细胞核抗原免疫染色出现在包含颗粒神经元前体细胞的外颗粒层(external granular layer,EGL)的外区,出现在 PD4-21 时。PD10 及以后,用 GFAP 免疫染色可以看到通过分子层和 EGL 垂直延伸的伯格曼胶质细胞突起。它们的胞体在 PD4 时对 Sox2 呈免疫阳性,随后在 PD10 时对 S100 蛋白呈免疫阳性,排列在浦肯野细胞层中。在 IGL 中稀疏地发现 Sox2 阳性细胞。在 PD90 时,少数细胞 NeuN 阳性,预示着成年神经发生的可能性。这些免疫组织化学结果表明,雪貂在出生后的生命过程中按照一定的顺序进行小脑皮质发生。浦肯野细胞单层排列和形态成熟的时间揭示了雪貂小脑相对缓慢的发育或成熟。