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小鼠神经系统中孤儿核受体RORα的免疫组织化学特征

Immunohistochemical characterization of the orphan nuclear receptor ROR alpha in the mouse nervous system.

作者信息

Ino Hidetoshi

机构信息

Department of Neurobiology (C1), Graduate School of Medicine, Chiba University, Chiba, Japan.

出版信息

J Histochem Cytochem. 2004 Mar;52(3):311-23. doi: 10.1177/002215540405200302.

Abstract

ROR alpha is an orphan nuclear receptor. A deletion mutation in the ROR alpha gene leads to severe cerebellar defects, known as the staggerer mutant mouse. Although previous in situ hybridization (ISH) studies have shown that ROR alpha is highly expressed in the cerebellum, especially in Purkinje cells, and in the thalamus, sufficient immunohistochemical (IHC) study has not yet been presented. I demonstrate here the IHC analysis of ROR alpha using a specific anti-ROR alpha antibody, in adult and developing mouse nervous system. ROR alpha immunoreactivity was observed in the Purkinje cell and molecular layers of the cerebellum. The co-localization of ROR alpha with calbindin D(28K) (CaBP) and parvalbumin indicates that ROR alpha-positive cells were Purkinje cells, stellate cells, and basket cells. In addition to the cerebellum, strong to medium ROR alpha immunoreactivity was found in the thalamus, cerebral cortex (mainly in the layer IV), dorsal cochlear nucleus (DCN), suprachiasmatic nucleus (SCN), superior colliculus, spinal trigeminal nucleus, and retina. The immunostaining was restricted in nuclei of neurons. Developmentally, ROR alpha immunoreactivity was observed in the cerebellum and thalamus from embryonal day 16 (E16). The distribution of ROR alpha immunoreactivity and ROR alpha mRNA hybridization signal was almost coincident. However, the intensity of hybridization signal was not always parallel to that of immunoreactivity.

摘要

RORα是一种孤儿核受体。RORα基因的缺失突变会导致严重的小脑缺陷,即蹒跚突变小鼠。尽管先前的原位杂交(ISH)研究表明RORα在小脑中高度表达,尤其是在浦肯野细胞和丘脑中,但尚未有充分的免疫组织化学(IHC)研究报道。在此,我展示了使用特异性抗RORα抗体对成年和发育中小鼠神经系统进行的RORα免疫组织化学分析。在小脑的浦肯野细胞层和分子层中观察到RORα免疫反应性。RORα与钙结合蛋白D(28K)(CaBP)和小白蛋白的共定位表明,RORα阳性细胞为浦肯野细胞、星状细胞和篮状细胞。除小脑外,在丘脑、大脑皮层(主要在IV层)、蜗背侧核(DCN)、视交叉上核(SCN)、上丘、三叉神经脊束核和视网膜中也发现了强至中等强度的RORα免疫反应性。免疫染色局限于神经元细胞核。在发育过程中,从胚胎第16天(E16)开始在小脑和丘脑中观察到RORα免疫反应性。RORα免疫反应性的分布与RORα mRNA杂交信号几乎一致。然而,杂交信号的强度并不总是与免疫反应性强度平行。

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