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Immunohistochemical Characterization of the Orphan Nuclear Receptor ROR in the Mouse Nervous System

Hidetoshi Ino

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

Correspondence to: Hidetoshi Ino, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan. E-mail: ino{at}med.m.chiba-u.ac.jp

ROR is an orphan nuclear receptor. A deletion mutation in the ROR gene leads to severe cerebellar defects, known as the staggerer mutant mouse. Although previous in situ hybridization (ISH) studies have shown that ROR 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 using a specific anti-ROR antibody, in adult and developing mouse nervous system. ROR immunoreactivity was observed in the Purkinje cell and molecular layers of the cerebellum. The co-localization of ROR with calbindin D_28K (CaBP) and parvalbumin indicates that ROR-positive cells were Purkinje cells, stellate cells, and basket cells. In addition to the cerebellum, strong to medium ROR 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 immunoreactivity was observed in the cerebellum and thalamus from embryonal day 16 (E16). The distribution of ROR immunoreactivity and ROR mRNA hybridization signal was almost coincident. However, the intensity of hybridization signal was not always parallel to that of immunoreactivity.

(J Histochem Cytochem 52:311–323, 2004)

Key Words: ROR • retinoic acid receptor • retinoid • cerebellum • thalamus • dorsal cochlear nucleus • immunohistochemistry

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