Coexistence of dopamine-beta-hydroxylase and activated protein–2 alpha in rat cerebellar Purkinje cells*☆

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Abstract

BACKGROUND:

Tyrosine hydroxylase and phenylethanolamine-n-methyl transferase expression coexist in Purkinje cells of the rat cerebellum. Numerous reports have also been published addressing whether dopamine-beta-hydroxylase (DBH) expression exists in cerebellar Purkinje cells.

OBJECTIVE:

To investigate the coexistence of DBH and activator protein-2α expression in rat cerebellar Purkinje cells.

DESIGN, TIME AND SETTING:

A cell morphological study was performed at the Institute of Neuroscience, Chongqing Medical University, China in May 2007.

MATERIALS:

Ten healthy Wistar rats, of either gender, aged 14 weeks, served as experimental animals. Rabbit anti-mouse DBH, goat anti-mouse activator protein-2α and rabbit anti-mouse β-actin (Santa Cruz Biotechnology, Inc., USA), horseradish peroxidase-labeled goat anti-rabbit IgG, FITC-labeled mouse anti-rabbit IgG, and Cy3-labeled mouse anti-goat IgG (Boster, Wuhan, China), were used in this study.

METHODS:

Immunohistochemical staining was used to measure the expression of DBH or activator protein-2α, with double-label immunofluorescence being employed to determine coexpression of both, in the cerebellum of 5 randomly selected rats. Western blot assay was utilized to determine the expression of DBH and activator protein-2α in the cerebellum of the remaining 5 rats.

MAIN OUTCOME MEASURES:

Expression, localization and coexistence of DBH and activator protein-2α in the cerebellum were measured separately.

RESULTS:

Immunohistochemical staining demonstrated that cerebellar Purkinje cells stained positive for DBH and activator protein-2α. Western blot assay also demonstrated DBH and activator protein-2α expression in the cerebellum. Double-labeling immunofluorescence showed the coexistence of DBH and activator protein-2α in cerebellar Purkinje cells.

CONCLUSION:

Norepinephrine and activator protein-2α coexist in rat cerebellar Purkinje cells.

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