Expression and localization of cytochrome P45011β,aldo mRNA in the frog brain

Minoru Takase, Kazuyoshi Ukena, Kazuyoshi Tsutsui

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

The present study is focused on biosynthesis of adrenal steroids in the frog brain. Employing RT-PCR method using total RNA from the adult Rana nigromaculata brain, we isolated a 419-bp fragment of cDNA encoding cytochrome P45011β,aldo, which catalyzes the final step of biosynthesis of the frog adrenal steroids, corticosterone and aldosterone. The deduced amino acid sequence of R. nigromaculata brain cytochrome P45011β,aldo shared a high homology (88.8%) with that of R. catesbeiana adrenal cytochrome P45011β,aldo. Southern blot analysis of the RT-PCR product confirmed the P45011β,aldo transcription in the frog brain without a clear-cut sex difference. Then, we analyzed the P45011β,aldo mRNA expression in different brain regions of the adult frog by RT-PCR method. The P45011β,aldo gene was transcribed in the telencephalon, diencephalon, midbrain, and cerebellum. The transcript level of the frog β-actin gene was relatively constant in all the frog samples examined. In situ hybridization analysis showed that the P45011β,aldo gene was transcribed abundantly in the cells throughout the frog brain, such as the pallium mediale in the telencephalon, the nucleus preopticus in the diencephalon, the stratum griseum superficiale tecti in the midbrain, and Purkinje cells in the cerebellum. These results taken together suggest that the frog brain synthesizes adrenal steroids, such as corticosterone and aldosterone.

Original languageEnglish
Pages (from-to)288-296
Number of pages9
JournalBrain Research
Volume950
Issue number1-2
DOIs
Publication statusPublished - 2002 Sep 20
Externally publishedYes

Fingerprint

Cytochromes
Anura
Messenger RNA
Brain
Diencephalon
Telencephalon
Steroids
Corticosterone
Mesencephalon
Aldosterone
Polymerase Chain Reaction
Cerebellum
Genes
Ranidae
Purkinje Cells
Southern Blotting
Sex Characteristics
In Situ Hybridization
Actins
Amino Acid Sequence

Keywords

  • CDNA cloning
  • Cytochrome P450
  • Frog brain
  • Gene expression
  • In situ hybridization
  • Neurosteroids
  • Reverse transcription-polymerase chain reaction

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Expression and localization of cytochrome P45011β,aldo mRNA in the frog brain. / Takase, Minoru; Ukena, Kazuyoshi; Tsutsui, Kazuyoshi.

In: Brain Research, Vol. 950, No. 1-2, 20.09.2002, p. 288-296.

Research output: Contribution to journalArticle

Takase, Minoru ; Ukena, Kazuyoshi ; Tsutsui, Kazuyoshi. / Expression and localization of cytochrome P45011β,aldo mRNA in the frog brain. In: Brain Research. 2002 ; Vol. 950, No. 1-2. pp. 288-296.
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