PUBLICATION
Activity and localization of 3beta-hydroxysteroid dehydrogenase/ Delta(5)-Delta(4)-isomerase in the zebrafish central nervous system
- Authors
- Sakamoto, H., Ukena, K., and Tsutsui, K.
- ID
- ZDB-PUB-011031-12
- Date
- 2001
- Source
- The Journal of comparative neurology 439(3): 291-305 (Journal)
- Registered Authors
- Sakamoto, Hirotaka
- Keywords
- neurosteroids; biochemical analysis; Western immunoblotting; immunocytochemistry; brain; teleost
- MeSH Terms
-
- 3-Hydroxysteroid Dehydrogenases/metabolism*
- Animals
- Blotting, Western
- Central Nervous System/cytology
- Central Nervous System/enzymology*
- Immunohistochemistry
- Male
- Neurons/cytology
- Neurons/enzymology*
- Pregnenolone/metabolism
- Progesterone/biosynthesis
- Steroids/biosynthesis*
- Zebrafish/anatomy & histology
- Zebrafish/metabolism*
- PubMed
- 11596055 Full text @ J. Comp. Neurol.
Citation
Sakamoto, H., Ukena, K., and Tsutsui, K. (2001) Activity and localization of 3beta-hydroxysteroid dehydrogenase/ Delta(5)-Delta(4)-isomerase in the zebrafish central nervous system. The Journal of comparative neurology. 439(3):291-305.
Abstract
Little information is available for neurosteroidogenesis in the central nervous system (CNS) of lower vertebrates. Therefore, in the present study, we examined the enzymatic activity and localization of 3beta-hydroxysteroid dehydrogenase/Delta(5)-Delta(4)-isomerase (3betaHSD), a key steroidogenic enzyme, in the CNS of adult male zebrafish to clarify central progesterone biosynthesis. Biochemical studies together with HPLC analysis revealed that the zebrafish brain converted pregnenolone to progesterone, suggesting the enzymatic activity of 3betaHSD. This conversion was significantly reduced by trilostane, a specific inhibitor of 3betaHSD. By using Western immunoblotting with the polyclonal antiserum directed against purified bovine adrenal 3betaHSD, a 3betaHSD-like substance was found in homogenates of the zebrafish brain. Immunocytochemical analysis was then undertaken to investigate the localization of the 3betaHSD-like substance in the zebrafish brain and spinal cord. Clusters of immunoreactive cell bodies were localized in the dorsal telencephalic areas (D), central posterior thalamic nucleus (CP), preoptic nuclei (NPO), posterior tuberal nucleus (PTN), paraventricular organ (PVO), and nucleus of medial longitudinal fascicle (NMLF). 3betaHSD-like immunoreactivity was also observed in somata of cerebellar Purkinje neurons. A widespread distribution of immunoreactive fibers was found throughout the brain and spinal cord. In addition, positively stained cells were restricted to other organs, such as the pituitary and retina. Preabsorbing the antiserum with purified bovine adrenal microsome resulted in a complete absence of 3betaHSD-like immunoreactivity. These results suggest that the fish CNS possesses steroidogenic enzyme 3betaHSD and produces progesterone. The present study further provides the first immunocytochemical mapping of the site of 3betaHSD expression in the fish CNS.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping